CA2396719A1 - 22 human secreted proteins - Google Patents

Abstract

The present invention relates to novel human secreted proteins and isolated nucleic acids containing the coding regions of the genes encoding such proteins. Also provided are vectors, host cells, antibodies, and recombinant methods for producing human secreted proteins. The invention further relates to diagnostic and therapeutic methods useful for diagnosing and treating diseases, disorders, and/or conditions related to these novel human secreted proteins.

Description

DEMANDE OU BREVET VOLUMINEUX
LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.

~~ TTENANT LES PAGES 284 A 436 NOTE : Pour les tomes additionels, veuillez contacter 1e Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS
THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE
VOLUME

NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME
NOTE POUR LE TOME / VOLUME NOTE:

Also preferred is a composition of matter comprising isolated nucleic acid molecules wherein the nucleotide sequences of said nucleic acid molecules comprise a panel of at least two nucleotide sequences, wherein at least one sequence in said panel is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from the group consisting of: a nucleotide sequence of SEQ
ID
NO:X wherein X is any integer as defined in Table 1; and a nucleotide sequence encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit with the ATCC Deposit Number shown for said cDNA
clone in Table 1. The nucleic acid molecules can comprise DNA molecules or RNA
molecules.
Also preferred is an isolated polypeptide comprising an amino acid sequence at least 90% identical to a sequence of at least about 10 contiguous amino acids in the amino acid sequence of SEQ ID NO:Y wherein Y is any integer as defined in Table 1.
Also preferred is a polypeptide, wherein said sequence of contiguous amino acids is included in the amino acid sequence of SEQ ID NO:Y in the range of positions beginning with the residue at about the position of the First Amino Acid of the Secreted Portion and ending with the residue at about the Last Amino Acid of the Open Reading Frame as set forth for SEQ ID NO:Y in Table 1-.
Also preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 30 contiguous amino acids in the amino acid sequence of SEQ ID NO:Y.
Further preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 100 contiguous amino acids in the amino acid sequence of SEQ ID NO:Y.
Further preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to the complete amino acid sequence of SEQ ID
NO:Y.
Further preferred is an isolated polypeptide comprising an amino acid sequence at least 90% identical to a sequence of at least about 10 contiguous amino acids in the complete amino acid sequence of a secreted protein encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit. with the ATCC Deposit Number shown for said cDNA clone in Table 1.

Also preferred is a polypeptide wherein said sequence of contiguous amino acids is included in the amino acid sequence of a secreted portion of the secreted protein encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table 1.
Also preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 30 contiguous amino acids in the amino acid sequence of the secreted portion of the protein encoded by a human cDNA
clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table 1.
Also preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 100 contiguous amino acids in the amino acid sequence of the secreted portion of the protein encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table l and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table 1.
Also preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to the amino acid sequence of the secreted portion of the protein encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit with the ATCC Deposit Number shown for said cDNA
clone in Table 1.
Further preferred is an isolated antibody which binds specifically to a polypeptide comprising an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of: an amino acid sequence of SEQ ID NO:Y wherein Y is any integer as defined in Table 1; and a complete amino acid sequence of a protein encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table 1.
Further preferred is a method for detecting in a biological sample a polypeptide comprising an amino acid sequence which is at least 90% identical to a sequence of at-least 10 contiguous amino acids in a sequence selected from the group consisting of: an amino acid sequence of SEQ ID NO:Y wherein Y is any integer as _.

defined in Table 1; and a complete amino acid sequence of a protein encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table .
1; which method comprises a step of comparing an amino acid sequence of at least one polypeptide molecule in said sample with a sequence selected from said group and determining whether the sequence of said polypeptide molecule in said sample is at least 90% identical to said sequence of at least 10 contiguous amino acids.
Also preferred is the above method wherein said step of comparing an amino acid sequence of at least one polypeptide molecule in said sample with a sequence selected from said group comprises determining the extent of specific binding of polypeptides in said sample to an antibody which binds specifically to a polypeptide comprising an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of:
an amino acid sequence of SEQ ID NO:Y wherein Y is any integer as defined in Table 1; and a complete amino acid sequence of a protein encoded by a human cDNA
clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table 1.
Also preferred is the above method wherein said step of comparing sequences is performed by comparing the amino acid sequence determined from a polypeptide molecule in said sample with said sequence selected from said group.
Also preferred is a method for identifying the species, tissue or cell type of a biological sample which method comprises a step of detecting polypeptide molecules in said sample, if any, comprising an amino acid sequence that is at least 90%
identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of an amino acid sequence of SEQ >D NO:Y wherein Y
is any integer as defined in Table 1; and a complete amino acid sequence of a secreted protein encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table 1.
Also preferred is the above method for identifying the species, tissue or cell .
type of a biological sample, which method comprises a step of detecting polypeptide molecules comprising an amino acid sequence in a panel of at least two amino acid sequences, wherein at least one sequence in said panel is at least 90%
identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the above group.
Also preferred is a method for diagnosing in a subject a pathological condition associated with abnormal structure or expression of a gene encoding a secreted protein identified in Table 1, which method comprises a step of detecting in a biological sample obtained from said subject polypeptide molecules comprising an amino acid sequence in a panel of at least two amino acid sequences, wherein at least one sequence in said panel is at least 90% identical to a sequence of at least contiguous amino acids in a sequence selected from the group consisting of: an amino acid sequence of SEQ ID NO:Y wherein Y is any integer as defined in Table 1;
and a complete amino acid sequence of a secreted protein encoded by a human cDNA
clone identified by a cDNA Clone Identifier in Table l and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table 1.
In any of these methods, the step of detecting said polypeptide molecules includes using an antibody.
Also preferred is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a nucleotide sequence encoding a polypeptide wherein said polypeptide comprises an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of: an amino acid sequence of SEQ m NO:Y
wherein Y is any integer as defined in Table 1; and a complete amino acid sequence of a secreted protein encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table l and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table 1.
Also preferred is an isolated nucleic acid molecule, wherein said nucleotide sequence encoding a polypeptide has been optimized for expression of said polypeptide in a prokaryotic host.
Also preferred is an isolated nucleic acid molecule, wherein said polypeptide comprises an amino acid sequence selected from the group consisting of an amino acid sequence of SEQ ID NO:Y wherein Y is any integer as defined in Table 1;
and a complete amino acid sequence of a secreted protein encoded by a human cDNA
clone identified by a cDNA Clone Identifier in Table 1 and contained in the deposit with the ATCC Deposit Number shown for said cDNA clone in Table 1.
Further preferred is a method of making a recombinant vector comprising inserting any of the above isolated nucleic acid molecule into a vector. Also preferred is the recombinant vector produced by this method. Also preferred is a method of making a recombinant host cell comprising introducing the vector into a host cell, as well as the recombinant host cell produced by this method.
Also preferred is a method of making an isolated polypeptide comprising culturing this recombinant host cell under conditions such that said polypeptide is expressed and recovering said polypeptide. Also preferred is this method of making an isolated polypeptide, wherein said recombinant host cell is a eukaryotic cell and said polypeptide is a secreted portion of a human secreted protein comprising an amino acid sequence selected from the group consisting of: an amino acid sequence of SEQ ID NO:Y beginning with the residue at the position of the First Amino Acid of the Secreted Portion of SEQ ID NO:Y wherein Y is an integer set forth in Table 1 and said position of the First Amino Acid of the Secreted Portion of SEQ ID NO:Y
is defined in Table 1; and an amino acid sequence of a secreted portion of a protein encoded by a human cDNA clone identified by a cDNA Clone Identifier in Table 1 and contained in,the deposit with the ATCC Deposit Number shown for said cDNA
clone in Table 1. The isolated polypeptide produced by this method is also preferred.
Also preferred is a method of treatment of an individual in need of an increased level of a secreted protein activity, which method comprises administering to such an individual a pharmaceutical composition comprising an amount of an isolated polypeptide, polynucleotide, or antibody of the claimed invention effective to increase the level of said protein activity in said individual.
The above-recited applications have uses in a wide variety of hosts. Such hosts include, but are not limited to, human, murine, rabbit, goat, guinea pig, camel, horse, mouse, rat, hamster, pig, micro-pig, chicken, goat, cow, sheep, dog, cat, non-human primate, and human. In specific embodiments, the host is a mouse, rabbit, goat, guinea pig, chicken, rat, hamster, pig, sheep, dog or cat. In preferred embodiments, the host is a mammal. In most preferred embodiments, the host is a human.

289.
In specific embodiments of the invention, for each "Contig m" listed in the fourth column of Table 5, preferably excluded are one or more polynucleotides comprising, or alternatively consisting of, a nucleotide sequence referenced in the fifth column of Table.5 and described by the general formula of a-b, whereas a and b are uniquely determined for the corresponding SEQ ID NO:X referred to in column 3 of Table 5. Further specific embodiments are directed to polynucleotide sequences excluding one, two, three, four, or more of the specific polynucleotide sequences referred to in the fifth column of Table 5. In'no way is this listing meant to encompass all of the sequences which may be excluded by the general formula, it is just a representative example. All references available through these accessions are hereby incorporated by reference in their entirety.

Gene cDNA CloneNT Contig Public Accession Numbers No. ID SEQ ID
ID
NO:
X

1 HTPFX16 33 1051320AA585439, N99013, AA585440, AL040768, AL044015, AL046147, AL042245, AL049007, AL043950, AL036500, AL043468, AL042700, AL044771, AL134123, AL046994, AL042712, AI535639, AL040414, AL040571, AL037341, AL046097, AI525556, AL045671, AI525316, AA585453, AL046150, AL044258, 228355, AL040856, AA585434, AI541510, AI546855, AL041577, AL043848, AL046914, AL044029, AL046850, AL037435, AL043570, AL079876, AI525328, AI541374, N99040, AL043201, AL040252, AL041459, AL045991, AL044201, AI556967, AI541514, C15189, AL043814, AI541523, AI526180, AL047593, AL044064, ' AL039316, 230131, AI546999, AL043923, AL043537, AL048647, AL043128, AI541534, AI525306, AL037335, AA585101, AL045994, AI535660, AI546945, AL03737.1, AI557731, AL043604, AI541365, AI557807, AI526140, AI541509, AI546828, AI541017, AI525431, AL037323, AL038532, AA585356, AI526194, AL043845, C16300, AI541317, AL044583, AIS47039, AL040329, AI526196, AI541535, AL041374, AL040082, AI557799, AI540967, AI54i508, AL040263, AI541307, AL039338, T11028, AI536138, AI557082, AI557262, AI525653, AI546899, AI526144, AI541205, D61254; 829445, AI535813, AI5S7787, AL040148, AL043627, AL041523,_.
AL044377, AL041730, 828735, AL038983, AL046392, AL044272, AL040090, AI546875, AI526184, AL040510, AL040625, AL045817, AL041142, AL041238, AL041133, AL041131, AL046330, AL040322, AL047183, AL041051, AL041292, AL047036, AL040119, AL047170, AL047057, AL041227, AL047219, AL040463, AL039915, AL043612, AL041197, AL040155, AL041346, AL040529, AL041096, AL047012, AL041358, AL041277, AL041163, AL041098, AL040621, AL043538, AL041324, AL040464, AL044162, AL041086, AL043496, AL041296, AL041233, AL043467, AL041159, AL045725, AL044186, AL041140, AL040193, AL044037, AL040168, AL040091, AL040128, AL040255, AL040342, AL040285, AL040332, AL040617, AL040553, AL045684, AL040745, AL043677, AL040370, AA585438, AL046442, AL040839, AL041752, AL040149, AL043775, D57491, AL044165, AI557084, AL043492, AL041602, AL045920, AL041278, AL038838, AL040253, AL044074, AL041635, AL045990, AL040458, AL044199, AL044187, AI525320, AJ239433, AL040294, AL041186, AL040052, AL042135, AL037727, T23985, AL040576, AI142134, AI546891, AL045753, AL044274, AI557796, AL079878, AL049018, AL040444, AL039744, AL045857, AL038822, AI525321, AL046327, AI541013, AL041168, AA585476, AL049069, AI557238, AL043444, AL041246, AL040472, AL040238, AL041955, AL041347, AI540920, C16305, AL038761, AL040075, T41289, T23957, AI541506, AL080031, AI557155, AI541345,.

AL045989, 829177, AL080075, AF221069, AF221068, AR017907, I13349, A91965, I66498, I66495, I66494, 166487, I66497, I66496, I66486, I66488, I66489, I66490, I66491, I66492, I66493, I66481, A83642, A83643,I66485,I66482,I66483,I66484, A83151, X81969, I05488, I61310, A60977, A91752, I08196, A32110, A58524, A58523, A68112, A68104, AR028564, AR067731, A06419, A21892, A23997, A84772, AR067732, A68114, A20702, A86792, A58522, AR062872, AR027319, AR037157, AR054109, A21895, A84776, A25909, A60985, A64973, A84773, A84775, A91751, AR062873, A47368, A43189, AR027318;

A84774, A43188, A60990, I19516, A05160, A08030, A20502, I19517, A89633, A89634, A60987, I63560, I63561, I63563, AR062871, I08776, AR009152, AR009151, E14304, AR002333, A81878, I25027, I26929, I44515, I26928, I26930, I26927, I44516, E 16678, I25041, A95096, A95106, A95105, A29109, A32111, A98767, A93963, A93964, I15353, A91750, A20700, AF082186, AR035975, AR035974, AR035977; AR035976, AR035978, I08051, A60961, A92133, I07249, AR068508, AR068510, AR068509, A63954, I91969, AR038762, I58322, I58323, AR003585, A76773, A22413, A38214, I56772, I95540, AR038855, A58521, A91754, AR031374, AR031375, AR020969, E12584, AR036903, A90655, AR008429, I01987, AJ244004, A85395, A85476, A49700, I44681, I18895, AJ244005, AJ244003, A92666, A92668, A9266'7, A92665, A98420, A98423, A98432, A98436, A98417, A98427, I63120, I19525, Y16359, D78345, X83865, A93016, A18053, M28262, AJ244007, I15717, I15718, E03627, I49890, I48927, A02712, A77094, A77095, I84553, A95051, I84554, A18050, A23334, A75888, 170384, A60111, A23633, AR007512, I08396, A60212, A60209, A60210, A60211, I00682, A11623, E00609, A11624, E13740, A11178, E01007, A10361, A62298, A35536, A35537, A02135, A04663, A02136, A04664, I08395, I06859, U94592, AR043601, A11245, I62368, X12660, A02710, E12615, AR035193, A07700, A13392, A13393, AR031488, I13521, I52048, A27396, AR027100, I44531, I28266, I21869, A70040, A82653, E16636, A24783, A24782, I03331, A95117, AF149828, I01995, AR031566, I60241, I60242, AR038066, A20699, E00696, E00697, E03813, AR027099, Y09813, AR051652, AR051651, A62300, and A25246.

2 HPJCP79 12 1097166AI131158, AI143233, AI084060, AI796995, AW024300, AI690136, AW068636, AA187976, AI990865, AA188104, AA702670, AI248604, AW241327, AI796740, AI638567, AA526922, AI147820, AA728795, AI955736, AI125679, AA036781, AI298768, AI653728, W58025, AI042104, AA903790, W57793, AI681520, N41467, AI148201, AI825715, AI982785, AL121341, AW068147, AA449957, AI917005, AA036838, AI125117, AW139372, AA536124, AI342363, 863264, H03741, AA451767, AI242453, AI928172, AI884809, AI536994, AI917991, AA833643, AW236630, AI769003, AI654929, AA364665, T88911, H06496, AI672523, AA343825, 845487, 866598, T79474, AA356796, AA135794, 807310, AA961256, AW 118743, W95716, AI339456, AI863802, AA135750, W95688, AA046714, 866752, 863216, AA046595, T84216, AW027460, AI203114, AI685956, AA112706, D20929, AI557697, AI525221, D42044, AL035413, AI637994, AI990544, AW071667, and AW235783.

2 HPJCP79 34 1035470W55896, W55856, AI033991, AI581098, AW068238, 876895, 886326, AW362820, 877060, AI028412, AI220844, AA862718, AA728770, AA040365, AA040364, T94138, D42044, AL035413, AI219616, and AW515266.

2 HPJCP79 35 1018167AI131158, AI143233, AI084060, AI796995, AW024300, AI690136, AW068636, AA 187976, AI990865, AA 188104, AA702670, AI248604, AW241327, AI796740, AI638567, AA526922, AI147820, AI955736, AA728795, AI125679, AI298768, AI653728, AA036781, W58025, AI042104, AA903790, W57793, AI681520, N41467, AI148201, AI825715, AI982785, AL121341, AW068147, AA449957, AI917005, AI125117, AA036838, AW 139372, AI342363, AA536124, AA451767, 863264, H03741, AI242453, AI928172, AI884809, AI536994, AI917991, AA833643, AW236630, AI769003, T08888, H06553, AI654929, AA364665, T88911, H06496, AA112706, AI672523, AA343825, T79565, 845487, AW068635, 866598, T79474, AA356796, AA135794, 807310, AA961256, AW118743, 824756, W95716, AA112705, AI339456, AI863802, AA135750, W95688, 807309, AA046714, 866752, 863216, AA046595, T84216, AW027460, AI203114, AA303951, AI685956, AI652260, D20929, AI557697, AI525221, D42044, AL035413, AI637994, AI990544, AW071667, and AW235783.

3 HWHS013 13 1115032AI628384, AI951331, AA625266, AI985375, AI247256, and AA41931 I.

_3 HWHS013 36 1045505T83059, AA419311, AI247256, AI628384, AI951331, and AI985375.

4 HSYAB05 14 1188478AA190860, AA355750, AA368604, AI674193, AL119522, AW384394, AW363220, AL119396, AW392670, AL134902, AL119443, AW372827, AL 119497, AL 119319, 299396, AL 119457, AL119399, AL119324, U46341, U46347, AL119484, AL119363, AL119341, AL119391, AL119355, U46346, U46350, U46351, U46349, AL119444, AL119483, AL119335, AL 119439, AL 119401, AL
119496, AL 134524, AL037205, U46345, AL042614, AL042896, AL134527, AI142132, AL134538, AL119418, AL043037, AL042980, AL042975, AI142137, AL042450, AL042965, AL042970, AL043019, AL134526, AL042542, AL042544, AI142139, AL042984, AL043029, AL042551, AL043003, AL119488, AL119464, AB026436, AR069079, AR054110, A81671, AR060234, AR066494, and AR043113.

4 HSYABOS 37 1030176AA355750, AL119522, AW384394, AW363220, AL119396, AW392670, AL134902, AL119443, AW372827, AL 119497, AL 119319, 299396, AL 119457, AL119399, AL119324, U46341, U46347, AL119484, AL119363, AL119341, AL119391, AL119355, U46346, U46350, U46351, U46349, AL119444, AL119483, AL119335, AL 119439, AL 119401, AL
119496, AL 134524, AL037205, U46345, AL042614, AL042896, AL134527, AI142132, AL134538, AL119418, AL043037, AL042980, AL042975, AI142137, AL042450, AL042965, AL042970, AL043019, AL134526, AL042542, AL042544, AI142139, AL042984, AL043029, AL042551, AL043003, AL119488, AL119464, AB026436, AR069079, AR054110, A81671, AR060234, AR066494, and AR043113.

HKIYI48 15 1212672AA421803, AA767372, AA746191, W90802, AA629842, AA723382, AA768200, AI379904, AI248375, W90746, 820049, AI651629, 860506, AI338678, AI470342, AI221377, AA325442, T09446, AW170290, and 245731.

5 HKIYI48 38 1042106820049, W90802, AA421803, and 245731.

5 HKIYI48 39 1039455AA629842, AA767372, AA746191, AI248375, AA723382, AA768200, AI379904, AA325442, W90746, AI470342, AI338678, and AA421803.

5 HKIYI48 40 1039454AA421803, AA767372, W90802, AA746191, AA629842, AA723382, AA768200, AI379904, W90746, AI248375, 820049, 860506, AI338678, AI470342, AA325442, 245731, and AI651629.

6 HARM138 16 1164812AW410706, AI748798, AA725637, AA402147, AI419313, AL049126, AI147283, _ . AI990726, AW410705, AI478369, AA020743, AI148856, AA284946, AA393789, AW236603, AA393884, AI889197, 856450, T27037, AI921312, AA394113, AI189667, AI948651, AA284766, AW172788, AA180305, AA402989, AI948636, AI866673, AW071781, H19479, AA284767, T27036, AW237208, AI500384, AI678354, 239780, AI826066, AI470278, H20936, AI270491, H84411, AA292332, H19561, AA285099, H20935, AI336345, 245818, 243720, AI541240, AW004032, AA020742, 815049, AI801457, F04685, AI290375, AW 135154, .

AI480181, AI611126, AI982513, AA179952, F21516, AA634385, 841485, and AI699619.

6 HARM138 41 1135023AA020742, 299396, AW392670, AL119497, AL119496, AW372827, U46341, AW384394, AW363220, AL119319, AL119457, AL119443, AL119324, AL119484, AL119363, AL119341, AL119391, AL119355, U46346, U46350, AI826066, U46347, U46351, U46349, AL134524, AL119483, AL119396, AL119335, AL119444, AL134920, AL134533, AL119439, AL042450, AL119522, AL134526, AL119399, AL042975, AL037205, U46345, AL042551, AL042614, AL134525, AL134538, AL042965, AI142132, AL134531, AL119418, AL042544, AL042970, AL043029, AL042542, AL043019, AL042984, AL043003, AL119464, ALI 19488, I25027, AR054109, I44515, I26928, I26930, I26927, I25041, I44516, AR035224, I85513, AR009152, AR009151, AR027099, I05393, A10617, AR028792, A94046, A94054, A01324, A01323, AR034783, I63120, AR067733, AR064322, AR064323, AR064320, AR064321, A32110, A94048, A94061, AR038307, AR038321, A49045, A83642, A83643, A70359, AR019094, I05430, A92666, A92668, I49890, A92667, A92665, AR028791, AR028793, AR019098, AR020199, AR020200, AR001287, AR020198, AR020197, AR029418, AR067734, AR067731, AR067732, AR029417, 189986, AR018924, AR018923, A48774, A48775, AR000006, AR015960, AR000007, AR015961, AR051652, A92081, A92080, A92077, A92078, A92079, A91752, A91751, I09121, AR068508, AR068510, AR068509, I91969, AR003585, I58322, I58323, A91754, A63067, A51047, A63064, A63072, A85308, AR031375, AR068507, AR068506, A60213, AR062871, AR051957, A44171, A29109, A32111, AR068550, A23373, AR068551, A49700, A60207, A60208, I58669, A58521, Y14971, AB026436, AR055065, A93444, A46342, A46343, A30600, AR055066, AR054110, A81671, AR032878, AR060234, AR066494, and AR069079.

6 HARMJ38 42 1038066AA020742.

6 HARMJ38 43 1031878AW410706, AI748798, AA402147, AA725637, AI419313, AL049126, AI147283, AI478369, AA020743, AI148856, AA284946, AA393884, AA393789, AW236603, AW410705, AI889197, T27037, AI921312, AA394113, AI189667, AA284766, AA180305, AW 172788, AA402989, AI990726, AI948636, AI866673, AW071781, H19479, AA284767, 856450, T27036, AW237208, AI500384, AI678354, 239780, AI470278, H20936, AI270491, H84411, AA292332, AI826066, AA285099, H20935, AI336345, H19561, 243720, AI541240, AW004032, 815049, AI801457, AI948651, F04685, AW135154, AI290375, AI480181, AI611126, AI982513, AA179952, F21516,-AA634385, 841485, 245818, AI699619, AI791349, Y17793, AF064854, and AI673327.

7 HTTIJ31 17 1165265AL046746, AL041706, AA488903, AW102955, AL120058, AI110760, AI471543, AI445582, AI631119, AI961232, AW024544, F28204, N23097, AA838140, AI888852, AA525409, AA568314, AI125167, AI925321, AL120343, AW410402, AA640305, AW410844, AA736713, AA679538,

8 PCT/USO1/01434 AW167374, AW265468, AA427470, AA449661, AA604333, AI890283, AA084439, AA573024, AL121039, AI702049, AL040921, AA515677, AI547110, AI538404, AI754257, AI572680, AF034176, AW021674, AI564209, AI792586, AI792498, AI908381, AI312267, AI445768, AI345166, AW148821, AI252611, AI431434, AW411147, AW021608, AA064983, AA533660, AA584360, AW 162288, AA502104, AW069110, AA610644, AC009516, AP000553, AC007227, AL022322, AL133448, 297630, AC006449, AC005529, AL034423, AC005399, AL031662, AC005484, AL031311, AL022165, 299716, AL008719, AP000692, AC003043, AC006285, L44140, AC004812, AC016025, AC006088, AC004263, AC005015, AC006006, AC005971, AP000501, AC005531, 283844, AC005102, AP000503, AL0224'76, AC005288, 284469, AC005695, AC005231, AC003688, AC004890, AC005527, AC004895, 298048, AC004703, AL034420, AC005500, Y14768, AL022326, AC004491, AL008582, AC000025, M63796, AL096791, 299128, AL121653, AC004560, U62317, AF207550, AC006312, AC004821, AC004033, AC006538, 298051, AC004851, AF196779, 295113, AC007676, AL022316, AL109628, AJ246003, AC005755, AC007055, AL049869, AC002314, AL009179, AC004253, AC005049, AC005081, AC004019, AC004922, AF111168, AL050318, AL133163, AP000505, AP000555, AL021154, AL022320, AC004150, AC005808, AL096701, U52112, AC004531, AC002418, AP000359, 297054, AC005331, AC002312, AC004000, AC006077, AC004999, AC007537, AC005913, AC006064, AC002126, AF053356, AC005280, AL035072, AC005031, AF001550, AL035405, AC004167, AC005412, AL023879, AC004099, U91326, AC003006, AC005785, U80017, AC007421, AL133243, AF109907, AC00211-5, AP000557, AF045555, AC007731, AC002492, AL035079, AL109627, AL021391, 298941, AL033521, AC004386, AC004972, AL022163, AC005821, AL035455, AC005071, AC006084, AL049759, AC004883, AC004876, AL031846, AC002~96, AC005519, AL109952, AC004858, AL050321, AC006509, AC006057, AC004659, AL049761, AC005225, U62292, AC005914, AC002404, AC002039, AC005209, AC007546, AC005619, AC005180, AL008710, AL031767, AC002553, AC005200, AC005358, U91327, AC005300, AL031281, AL135744, AC004887, AC005228, AC000353, AL133245, AC002477, AC007384, AF003626, AL035086, U85195, AP000691, AC003689, AL035422, AB023048, 293017, AL031255, AC006261, AC005562, AC007666, AF205588, AC004584, AC005736, AL022336, AC007308, AL022313, 285986, AC005523, ' AL035659, AC002563, AL021453, AC007435, AL022315, AL008729, AL035249, AE0006S8, AC006001, AL022323, AC004638, AC004477, AL117258, AL117330, AP000356, AC007993, AC005089, U78027, AC003109, D87675, AL049776, 297056, AC007919, AC005839, 281364, AL031848, AL031670, AF134726, AC005694, AC002310, AD000092, 282244, AL035587, AC004832, AC005250, AC010205, AL031427, AL049636, AC004878, AB023049, AL008716, AF111167, AL121915, AL049694, AP000116, AC004973, 293241, AL031657, AC004134, AC007277, AC005086, AC005911, AL121655, AC002395, AL080243, and U91321.

7 HTTIJ31 44 977169 AC009516 and AP000553.

8 HHTLB76 18 1138860AA829036, AI061313, AA449997, AI733856, AW328331, AI609972, AA535216, AI612142, AI755214, AI610941, AL079734, AI279417, AI7S4S67, AI5832S2, AA704393, AA410788, AI754105, AI912401, AI249365, T74524, AI962030, AL041375, AA584862, AA483606, AI926033, AI380617, AW237905, AA225406, AA570740, AW272294, AW327624, AA838091, AA169245, AA420546, T57767, AI884383, H07953, AI491765, AI635028, AA502991, AI446623, AI114557, AA349193, AA568204, AW265359, AI792521, AL037714, AI859438, AA469327, AA829065, AI251576, AA862312, AW301771, AA228778, AA828592, AA492584, AA297666, AIS81193, AI628859, AI249688, AI457596, AI457389, AA534064, AA812684, AA552989, AJ229041, AL050317, AC008179, AC005874, AF134471, 297181, AJ003147, U82695, AC004526, AC005529, AL022162, AP000245, AL022098, AF031078; AF030876, AC006263, AL031295, AC007228, AC003037, AL049839, AC004S34, AC007401, AC006427, 285987, AP000014, AC002369, AC002123, AP000036, AC002378, AC004970, AL133238, AF031076, AC006080, AC005538, AL022336; AC001231, AP000206, AP000128, AC005527, AL023807, AP000098, AL031289, AC005520, AL078638, AC.003035, .

AC005060, AC005533, AC003684, AL022337, AJ229042, AL031121, AL008729, AL031681, AL009031, AC002326, AP000501, AC002302, AF001550, AC004554, AC005284, 293930, AC005192, AC004887, AF011889, M63544, AC004890, 284466, AL080243, AC007073, AC007376, AL034402, M63543, AP000330, AL031597, AL096703, U91321, 298941, AC005914, AL049869, AC005531, AC007919, AC005799, AF111169, AC006928, AC003657, AL031123, 297353, AP000032, D86995, AC002347, AL078477, 298751, AL022163, 285994, AP000260, AC004966, AC002070, AP000696, AL009181, AC004778, AC005871, AC007450, AL031737, AL034429, AC007298, AC005291, AC006961, AC007263, AL031664, AC004125, AP000099, AC005859, AL034384, AC005722, AC004967, AC006101, AC003663, AP000212, AP000134, AP000030, AC004551, AP000466, AC004685, AP000689, AC005694, AC016831, L44140, AC005703, AL133244, AC008372, AC005036, AC007731, AC007057, AC004020, AC006211, AP000013, AC004865, AC007129, AC006992, AC004674, AL121653, AP000295, AC007917, AC004787, AF003529, U95739, 298304, AC002549, AC007051, 283826, AL022326, AJ251973, AL031733, AC004816, AC006257, AL049795, AC008041, AC005822, AC005500, AC004895, AL021939, AC005295, 282195, AC004216, AC005699, AC005081, AC004453, AC006252, AC002067, AF196969, AC002351, AC005377, AC007011, AC007207, AC007276, AL 122003, AP000103, AP000109, AP000041, AC005702, AL033525, AL049712, 295331, 285996, AC005274, AL133445, AP000247, AL035420, AC007993, AP000121, AP000053, AP000168, AL022320, AC005697, AL008719, AC005255, AC002288, AP000111, AP000043, AC007151, AL133163, AC002365, 284572, AL008730, AC002492, AL031577, AL031297, AL132642, 283836, AP000500, AF196970, AP000263, AL031848, AC004682, AC002045, AP000688, AF 121782, AC008115, AF015262, AC012384, AC005245, AB003151, AF003626, AL078593, AC006203, AC007684, AJ229043, AC002456, AC004687;

AP000056, AC006011, AC004143, 298050, AC003003, AL033397, AC006130, AC005823, AL034549, 282198, 298044, AC005399, AF091512, AC006505, and AC006160.

8 HHTLB76 45 1136207AL134524, AI525316, AA829036, AI733856, AI061313, AA535216, AI609972, AI612142, AI279417, AW328331, AI610941, AL079734, AI583252, AI912401, AA483606, AI380617, AI249365, AW237905, AA570740, AI962030, T74524, AA169245, AA704393, AI755214, AA838091, AA225406, AI251576, AI792521, AI754567, AI884383, AA568204, AW272294, AI754105, AA584862, AL037714, AI635028, AA502991, H07953, AI583466, AW265359, AA862312, AW301771, AA829065, AA828047, AA828592, AL041375, AI306232, AI890971, AW274191, AA631517, AA013168, AI114557; AI281431, AI961983, AI417469, AI053696, AI628859, AA558404, AI280266, AA629894, AA629902, H53284, AW021116, AW439703, AA019973, AI310464; AI793172, AI793209, AJ229041, AL050317, AR038762, AF134471, AC008179, U82695, AP000245, AF031078, AF030876, AJ003147, AL022098, AC005874, 297181, AP000036, M63544, AF031076, AL031295, AC002369, AC006263, M63543, AL049839, AC004534, AC002123, AC005913, AC001231, 285987, AC006427, AC007228, AC004526, AL132642, AL133238, AP000206, AP000128, AC005538, AL009031, AC002326, AC006080, AC003684, AC003037, AL031289, AF011889, AL022337, AL078638, AC007263, AC005192, AL031737, AC004890, AC005060, AL031597, AC005036, 283826, AC003035, AC007073, AC007401, AL008729, AC005533, AF111169, AC005529, AP000330, AC007130, AJ229042, AL023807, AL049869, AC005699, 285994, AC004966, AC006211, AC002302, AC004685, AC004554, 293020, 284466, AL031733, AL022336, AC004967, AP000260, AL049712, AC007376, AC005859, AL009181, AL034384, D86995, AC006101, AL096703, AC005284, AL080243, U95739, AC007917, AJ229043, AC006515, AF003529, 295331, AC007057, 285996, AC005799, AC004887, AC005694, AC007919, AP000099, AL049795, AP000030, AC006961, AC005527, AF015262, AC007298, AP000240, AC004778, AC002378, AL034429, AC007051, 298941, AC002347, AL022162, AC004787, AC005871, AC008115, AP000466, AP000696, AL035461, AP000212, AP000134, AF003626, AC005377, AL022163, AF001550, AF196970, AC007878, AP000032, AC005971, AP000689, AC007276, AC007684, AC005386, AL078593, AC008372, U91321, AC006928, AC007151, AL031681, AP000694, AC007129, 295113, AC003657, AB003151, AC004970, AC004551, AC005531, AC005291, AL031123, AP000295, AC005295, AP000103, AC004020, AP000109, AP000041, AL008719, AC002351, AC005722, AF196969, 268287, AC006257, AC016831, AC002549, AP000697, AC005245, AC009044, AL133244, AP000098, AC005255, 297353, 298044, AC005822, AC005914, AL031121, AC006974, AC006160, AL049636, AL022326, AF099810, AL034402, AC002045, AC002067, AP000501, AC004816, AF091512, U07000, AC005703;

AC007011, AC008041, AP000155, AC003003, AL122003, AC007731, AC006011, AC007157, AL121757, AC004674, AC006075, AC005399, AL022316, 282195, AP000263, AC006992, AL021977, AL031120, AC007450, 298304, AC002070, AC005071, AL035089, L44140, AC006953, AP000247, AP000688, AL121653, AC004865, AC004963, AC005500, AL078584, AC003098, AP000201, AP000013, AC007262, AP000143, AC004682, AL049553, AP000097, AL133445, AC004453, AC005786, AP000493, 298200, AC003663, AC004143, AC005520, AC005081, AL078477, AC006238, AP000111, AP000043, AL033525, and AP000555.

9 HTEGF16 19 1160908AA203365, AI813708, AW300554, AW026370, AI654923, AA252931, AW299408, N70033, AI859872, AA977420, AA629258, N46021, 877192, AI659888, AA252888, H11937, AA149537, AI024609, N69894, AI768599, AI420563, AI472810, AI810949, AA489009, N98886, AI698854, AA364127, 238327, AA488535, AI187721, AA488402, AI803900, AA309914, and AW612414.

9 HTEGF16 47 1049385877192, H11937, N46021, N69894, N70033, N 98886, AA203365, AA252888, AA252931, AA977420, AA488402, AA488535, AA629258, AI024609, 238327, AI420563, AI654923, AI698854, AI768599, AI803900, AI810949, AI813708, AW026370, AW299408, AW300554, and AW612414.

9 HTEGF16 48 952104D51799, D80439, AA305409, D80212, D59859, C14331, D59619, D80210, D80240, D80166, D80157, D81030, C14014, C14389, D51060, D80219, D80164, D51423, D80133, D80253, D51759, AA514186, D80268, D80195, D59610, D80022, D80366, D58283, D80188, D80247, D80391, D80248, D45260, C06015, D59787, D59502, D59889, D59467, D59275, D80038, D80043, D80227, D80045, D81026, D57483, D80196, D80269, D80024, D59653, D51022, D50979, D80522, D50995, AA305578, C05695, D59927, C15076, C14429, D80241, D80193, D80302, C03092, AA514188, D80251, D80378, D51103, D81111, AW 177440, AW 178893, D80064, AW360811, D59373, D80949, AA809122, D58246, H67854, T11417, C14344, F13647, C75259, D51221, AW377671, AW375405, AI525920, AI525917, H67866, AI525923, T03269, T03116, AA514184, AW360817, AW360834, D80258, C14227, D59503, D59317, D80014, D59474, D59695, AW366296, C14973, D51079, AW178906, AI535686, AW360844, D80228, C14407, AW 179328, T48593, D59551, D58101, AW375406, AW378534, AW179332, AW377672, AW 179023, AW 178905, AW177731, C14957, AW378528, AWi78762, AW179019, D60010, AW378533, AW378532, C14046, AW177501, AW177511, AI525235, 221582, D60214, AW 179020, AI525215, AI525227, AW377676, AI557774, AW352171, AW360841, AW352170, AW178907, AW 177733, AW 178908, AW 179024, AW352120, AI525912, AI525925, D51250, D59627, AW 178914, AW 178774, AW 176467, AI525242, AW178986, AA285331, AW 177505, AI557751, AI525222, AW 178775, D51053, AW367950, AW 178909, AW 177456, AW 179004, AW 179329, AW
178980, D45273, C 14298, AW 178754, 230160, AW 179018, D51213, AW352158, C16955, D80168, AW378542, AW352117, C05763, 233452, T02974, 217814, AW178781, AW378539, AW 178911, AW378543, AW378525, AW378540, AW352163, AW360855, AW 177734, AI525237, H67858, AW 179009, AW 179012, C04682, AW 177728, AW 177722, AI525238, T02868, AI910186, AW369651, C139S8, AI525928, N66429, C14077, D80314, D31458, AI525216, D50981, F13796, AW 177508, AI525228, AW 177497, T03048, AI525239, AR060385, A62298, AR018138, AR008278, A82595, A84916, A62300, AB028859, AJ132110, AF058696, AB002449, I50126, I50132, I50128, I50133, I14842, AR054175, AR008277, AR008281, AR016514, A63261, X67155, Y17187, AR060138, A45456, Y17188, A94995, D26022, A26615, AR052274, A43192, . Y12724, A43190, AR038669, A25909, AR066488, Y09669, AR066487, A70867, A67220, D89785, A78862, D34614, A30438, AR062872, AR008443, I79511, AR016691, AR016690, U46128, A64136, A68321, D88547, 182448, D50010, X68127, X82626, X72378, AR008408, AF123263, AR032065, AR060133, and AR025207.

HISET33 20 1102795AL134524, AI142134, AL045327, AL134110, AL045328, AL038983, AL038878, AL047163, AL119324, AL042898, AL037727, U46344, AL039432, AW372827, AL135012, AL037295, AL038838, AL048677, AL037343, AL039643, AI318479, AL038651, AIS47295, AL119457, D29033, AL037436, AL037335, AL037323, AL049018, AL119399, AL037443, AL038532, AW392670, AL038822, AL042420, AL047611, AL042655, AI431323, AL045494, AL045891, AL042523, AL038761, AL037435, AL042741, AL040472, 299396, AL043941, AL043321, AL044125, AL043923, AL043814, AL047012, AL041238, AL044186, AL040617, AL043845, AL041347, AL042931, AL040576, AL040193, AL045753, AL041955, AL040463, AL047170, AL044037, AL041635, AL040294, AL044064, AL041459, AL041577, AL038024, AL043089, AL044162, AL047219, AL040625, AL048657, AL045684, AL041752, AL046850, AL040768, AL045671, AL046994, AL046914, AL048714, AL039360, AL038745, AL119443, AL042519, AL043496, AL040052, AL042802, AL042508, AL043538, AL040444, AL040621, AL040464, AL042488, AL046356, AL040510, AL043467, AW363350, AL043677, AL040839, AL043492, AL041602, AL044074, AL041730, AL041523, AL043627, AL041374, AL043848, AL043570, AL047183, AI431307, AL042135, AI623302, AI432644, AI431316, AI431238, AL042515, AL046442, AL041324, AL041133, AL042533, AL039316, AI432666, AL041098, AL040322, AW384394, AL042853, AL046392, AL042832, AI431235, AI431315, AL040119, AW363220, AL044272, AL044258, AL043091, AL037341, AL042468, AL134920, AL041096, AL119396, AL042729, AL042096, AI431246, AL043166, AL042842, AL042787, AL041168, AL119497, AL038040, AW081103, AI432653, AI431230, AI431321, U46350, AL041163, AL041.1~59, AL119483, AL045817, AI431257, AL045920, AL040148, AL119418, AL079852, AL047057, AL040207, AI432654, AL045326, AI432650, AI432677, AL040458, AL044187, U46349, AL043278, U46351, AL043295, AL038041, AL119335, AL041296, AW084068, U46341, AI431328, AL041358, AL041292, AL119319, AL040571, AL045990, AL047675, AL119341, U46346, AI431353, AL119522, AI431312, AI432656, AI431231, AI432655, AR066494, AR064707, AR023813, AJ238010, A93923, A93916, D 17247, A93931, AL 133053, AR060234, AL122101, AL133074, AF019249, AL133049, Y17793, A85203, A81671, AL133082, AL133076, AL133068, AR069079, AR054110, and AB026436.

HISET33 49 977186AW392670, 299396, AL119396, AL042965, AW372827, AW384394, AL119497, AL119401, AL119443, AL119483, AW363220, AL119418, AL134524, U46350, AL134533, U46346, AL119335, AL119522, AL134902, AL134920, AL119341, U46349, U46341, U46351, AL119496, AL042896, AI142132, AL119399, AL134531, AL134538, AI142139, LJ46345, A81671, AR060234, AR066494, AR069079, AR054.110, and AB026436.

1 I HTFMX90 21 1055914AL134524, AL042420, AA515728, AA829036, AI279417, AW237905, AI887235, AI358712, AI380617, AI912401, AW419389, AI926728, AA706495, AI244127, AA225406, AI755214, AI754105, AA572813, AI754567, AW026305, AI421950, AI904840, AI419337, AA704393, AI090377, AI224583, AA573067, AA640430, AI859438, AA502991, AI612142, AL118925, AA640410, AL031428, AP000245, AP000128, AP000206, AL109627, AC005274, AF139813, AC005740, AC004228, AL034549, 282206, AL050331, AC005670, AL109801, AL079303, AP000503, AF134726, AC005921, AP000696, AC005520, AC003669, AC005071, AC006480, AC007371, AC005228, AC006530, AC0072,16, AC002477, AC006449, AL031589, AC007052, AC007363, AP000694, AC002301, AC002425, AP000113, AP000045, AL021154, AC004139, AC003982, AL049780; AL021155, AC005668, AC005412, AC003101, AP000211, AP000133, AC006064, AC004827, AC005399, AL121652, AL031311, AB014078, 273417, AC004020, AC008179, AP000697, AC005899, AL049697, AL133244, AL031664, AL022069, U80017, AL096791, AC004821, AC004859, AL022334, AL137100, AC004797, AC002549, AC007421, AC006388, AC002365, L44140, AC006501, AL023803, AC002429, AC003663, AC004805, AP000514, AL021394, AC007707, 284469, AC004181, AC002565, AC002073, AC004765, AB020868, AL034420, AJ003147, U95742, AC006312, AC005031, AC004774, AC004841, AC007731, AC005011, AF042484, AP000497, AC005500, AL133396, AC005039;

AL121655, AL035587, AC000353, U91321, AL049831, 282195, AC006441, AL049839, AC005920, AL033521, Y14768, AC002045, AL022399, AC004905, AC006211, AL049759, AL031602, AL133448, AC006061, AC004079, 293241, AL031228, AC006487, AC007773, 282194, AL035423, AC005701, AL024507, AP000505, AC007842, AP000240, AC005696, AL078477, 282172, AC004891, AC006581, AC005295, AL034554, AF165926, AC002115, AF064861, AL034548, AC005799, AC004975, AC005536, AC005585, 284486, AL031737, AL031670, AL035410, AC007298, AF111168, AL049871, AP000098, AC002302, AC007917, AB023048, AC005913, AC006597, AC002401, AL078583, AC002558, AL133353, AP000563, AC007041, AC008125, AL031666, AL079304, AL133245, AP000114, AP000046, AL009181, 282215, AC005231, 285986, AB000876, AC007686, AC005049, AL035683, M89651, AL035089, AL031283, AC005901, AP000557, AL133243, AC005409, AC002314, AC005778, AC004383, AC006115, AC007243, AL096775, AC008372, 293017, AC004972, U91326, AF124523, AL109798, AC006111, AC005939, AP000022, AL035413, AC005089, AC010205, AF064857, AC005411, AC005592, AC005225, AP000501, AC002316, AF196779, AC007151, AC005940, AL035088, AC006023, 297056, AC002080, AC002449, U95743, AC007386, AL031774, AL121748, AC004491, AC007226, AL021393, AL109623, AC005058, AC004125, 293244, AL022316, AC007868, AC005086, AC000063, and AL035407.

11 HTFMX90 51 1052131AA515728, AA829036, AI279417, AW237905, AI887235, AI358712, AI380617, AI912401, AI244127, AI926728, AI755214, AA225406, AW419389, AI754105, AA572813, AI754567, AA706495, AW026305, AI421950, AI904840, AI419337, AA704393, AI090377, AI224583, AA640430, AI859438, AA573067, AA502991, AA640410, AL118925, AI311647, AI361090, AC007216, AC005274, U91321, AC007363, AP000128, AP000206, AC000353, AC006449, AP000245, Y10196, AC007052, AF111169, AL031428, AC006480, AC005740, AP000503, AC004228, AF139813, AL109801, 282206, AC003669, AP000696, AC003101, AL079303, AC005670, AL031589, AL109627, AC005412, AC006530, AC005921, AL049697, AP000694, AC006501, AC005520, AF134726, AL049780, AC004139, AC002301, AC003982, AC006061, AC006064, AC003663, AC004765, AL035410, AC002477, AP000211, AP000133, AC004827, AL031311, AC002425, AC005668, AC004020, AC005039, AP000113, AP000045, AL133244, AC005228, U80017, AL034549, AL022334, AP000697, AC007277, AC004821, AC002365, AF042484, AL031664, AC004805, AC006388, U95742, AC006211, AL021155, AL022069, AL034420, AC005399, AC004905, AL021154, AC005778, AC004797, AB020868, AJ003147, 293241, AC002395, AL121652, AL096791, AC008179, AB014078, AC005071, AP000514, 284469, AP000497, AC007421, AC007707, AL049831, AL121655, AL035407, 293244, AL078477, AC002073, AC002045, AF111168, AC006312, AL031737, AC002565, AC005031, AC004774, AB023048, AL133396, AC004181, AL050331, AL049759, AC004859, AL031602, AC005913, AC006581, AC004841, AC002558, 273417, AL031228, AL035587, AC005011, AC005701, AC006115, 282195, AC005920, AC005899, AC007842, AL023803, AC007773, Y14768, L44140, AC005696, AP000240, AL022399, AC002316, 282194, 282172, AL033521, AF165926, AL133448, AC007055, AL024507, AC005536, AC005585, AC004975, AF196779, AC002429, AL035423, AC006441, AF064861, AL049871~, 298946, AC005295, AP000505, AC007371, AC004125, AC007731, AC002115, AL133353, AC008372, AL031666, AC005500, AP000022, AL009181, AC005005, AC007298, 285986, AL133243, AC004891, AC006088, AL031283, AC005821, 282215, AC005049, AC005409, AC004079, AL137100, AP000098, AR036572, U91328, AC002302, AL021394, AC005088, AC008125, AC005690, AC004491, AL031670, AF124523, AL033392, AC010205, AC005411, AL109798, AC006487, AP000365, AP000114, AP000046, AC007243, AC002401, AE000658, AL049757, AC005231, AC004383, U91326, AC004972, AL079304, AC006111, AP000501, AB000876, AC002314, AC007227, AL121748, AC007041, AC005086, AF064857, AL049839, 293017, AL035683, AL022238, AC006511, AC005901, AC004938, AC007677, AL078583, AC003041, AC008101, 298742, AL139054, AC007917, AC003035, AC007386, AC002544, AC007066, AL0313.68, AC007686, AL022328, U85195, AC007221, AC004929, and AC002080.

12 HE8FD93 22 1091113AC007350.

12 HE8FD93 53 1046606AA142989, AA649871, AA150979, AA730301, AI423201, and AC007350.

12 HE8FD93 54 948832 AC007350.

13 HSUMA53 23 1183531AW375619, AW375590, AI954621, AW362044, H98087, AW205215, AW205991, AI361588, AW193731, AA588837, AA534307, AI631442, AW291638, AW408128, AI097266, AA731997, H89823, 898300, W01922, AI656446, 898059, AI432644, AI623302, AI431255, AI431337, AI431351, AI432654, AW128900, AI432658, AI432674, AI432661, AI431346, AI791349, AI432675, AI431353, AI431347, AI432653, AI431230, AI431328, AI431354, AI432655, AI431310, AI431312, AI431241, AI431345, AW081103, AI432677, AI431254, AI432651, AI432647, AI432665, AI431357, AI432673, AI432649, AI431243, AI492519, AI431330, AI432672, AI431248, AI432662, AI432676, AW 128846, AI431340, AW 128897, AI432664, AI432650, AI431307, AI431316, AI492520, AI432643, AI492509, AI431751, AW128884, AI432657, AI431352, AI431247, AI431356, AI492510, AI431308, AI432645, AW 129223, AB006624, Y17793, AF064854, AF019249, and AF048686.

13 HSUMA53 55 951341 AA380314, AA374807, and AB006624.

13 HSUMA53 56 898943 AW375619, AW375590, AI954621, AW362044, H98087, AW205215, AW205991, AI361588, AW193731, AA588837, AA534307, AI631442, AW291638, AW408128, AI097266, AA731997, H89823, 898300, W01922, AI656446, 898059, . AI432644, AI623302, AI431255, AI431337, AI431351, AI432654, AW128900, AI432658, AI432674, AI432661, AI431346, AI791349, .

AI432675, AI431353, AI431347, AI432653, AI431230, AI431328, AI431354, AI432655, AI431310, AI431312, AI431241, AI431345, AW081103, AI432677, AI431254, AI432651, AI432647, AI432665, AI431357, AI432673, AI432649, AI431243, AI492519, AI431330, AI432672, AI431248, AI432662, AI432676, AW 128846, AI431340, AW 128897, AI432664, AI432650, AI431307, AI431316, AI492520, AI432643, AI492509, AI431751, AW128884, AI432657, AI431352, AI431247, AI431356, AI492510, AI431308, AI432645, AW 129223, AB006624, Y17793, AF064854, AF019249, and AF048686.

14 HEOQX60 24 1035205AA614014, AW084519, AA903338, AI244442, AI342240, AI962752, AI717991, AW139714, AA354504, AI660761, AW207518, AW402742, D20168, AA380395, T29020, AW138271, AI219797, AI041118, F19235, AI798637, F16699, AI201892, AL047399, AI808637, AI564067, AW393736, AW393737, AA748165, AA918804, AL041067, AA282129, AL041068, AA282128, AA811883, AI921541, AW393733; 299396, AL119443, AW392670, AW372827, AL119497, AL119418, AL119457, AW384394, AL119496, AL119483, U46341, AW363220, U46349, AL119324, U46346, U46351, AL119391, AL036418, AL038837, AL134524, AA631969, AL119335, AL119399, AL119484, AL037051, AL036725, U46350, AL119319, AL042544, AL134902, AL119363, AL119341, AL119444, AL119396, AL119439, U46347, AL134518, AL119355,.AI142132, AL119522, AL037205, AL042614, AL038509, AL036858, AL036924, AL039074, AL119401, AL134528, U46345, AL042965, AL042975, AL134525, AL134538, AL119464, AL042970,.AL042450, AL042984, AI142137, AL042551, AL037082, AL043019, AL043029, AL042542, AL037639, AL037094, AL036196, AL037526, AL043003, AL119488, AL036767, AL038851, AL037085, AL037077, AL036268, AL036190, AL038520, AL037615, AL036998, D30758, AR060234, AR066494, A81671, AR023813, AR064707, AR054110, AR069079, and AB026436.

14 HEOQX60 57 918920 AI253192, AI913253, AI597595, AA713495, AA972065, AA135967, AI928486, AA621901, AI261821, N87925, AA767100, AA427998,.

AW375405, C14331, AA514186, D80188, D58283, D59275, AW366296, D57483, D80253, D80166, C14014, D51060, D80024, AA305409, C14389, D59859, D80043, D80366, D51799, D80248, D51423, D59619, D80210, D80240, D50979, D81030, C14429, D80212, D80022, D80219, D80195, D59467, D80391, D80164, D59610, D59787, D80227, AA305578, D59502, D80133, D80522, D81026, D80269, D5.9889, D80196, D51022, D80268, D50995, D59927, D80251, C15076, D80038, D80439, D80193, D80045, D80241, AA514188, AW360811, D80378, AW177440, D80247, D80302, AW 178893, D59373, AW377671, AA809122, C75259, AI557751, AI905856, T03269, T48593, C05695, AW 178906, AW 179328, D51103, AW360844, D51759, AW360817, D80157, AW375406, AW378534, AW179332, AW377672, AW 179023, AW 178905, AW378532, AW177501, AW177511, AW352171, AW377676, AW178983, AW352170, AW 177731, AW 178907, AW378528, AW 178762, AW 179019, AW
179024, AW352117, AW176467, D51250, D80134, AW360841, AW177505, D80132, C14407, D58253, AW 179020, AW 178775, AW 178909, AW 177456, AW 179329, AI535686, AW 178980, AW 178914, AW
177733, AW 178908, AW 178754, AW
179018, AW352158, D52291, AW367967, AW369651, D59653, C06015, AW360834, AW 178774, AW352120, AW179004, D45260, AW179012, AW352163, AW378525, D51079, F13647, D80949, AW378543, D59627, C14344, AW 177728, D80258, H67854, AW 179009, AW 178911, AW 177722, AW367950, AI910186, AW378540, D80168, AW352174, C03092, D80064, H67866, AW178781, C14298, C14227, D59503, T11417, AI525923, C14957, D59317, D58101, D81111, AW177734, AI525917, D58246, AW178986, D59474, 221582, D51221, AA514184, D80228, T03116, AW 177508, AW 177723, D80014, AI525920, C14973, AI525912, AW378533, AW177497, D60010, D51213, D45273, AI525235, D59551, AI525227, D51097, C14046, D60214, AI557774, N66429, T03048, AI525215, C16955, AI525242, AA285331, AW378542, AI535959, AI525925, AI525222, AW378539, AI525237, C05763, 233452, T02974, AW360855, I25810, X99477, A62298, A84916, A62300, A82595, AR018138, Y17188, AJ132110, AB028859, AR008278, AF058696, AR016808, A30438, Y17187, A94995, AR060385, X82626, AB002449, X67155, D26022, Y12724, A25909, AR016514, A67220, D89785, A78862, D34614, AR008443, I50126, I50132, I50128, I50133, D88547, AR066488, AR060138, A45456, A26615, AR052274, A43190, AR038669, Y09669, X68127, A43192, AR025207, AR066487, AR008277, AR008281, D50010, I14842, AR054175, U46128, AR016691, AR016690, AR066490, A63261, U79457, AR008408, I18367, AR062872, A70867, 282022, D13509, AR060133, AB012117, A64136, A68321, I79511, A44171, A85396, D88507, AR066482, AF123263, A85477; I19525, A86792, AR032065, X93549, and AR008382.

15 HFXDI56 25 1218288T85435, T85337, AA904610, AI220608, AL117340, AP000080, and X55499.

15 HFXDI56 58 1218744T85435, T85337, AA904610, AI220608, AL117340, and AP000080.

15 HFXDI56 59 1035197AL117340.

15 HFXDI56 60 609313 T85435, T85337, AA904610, AI220608, AL117340, AP000080, and X55499.

16 HCLTHQ40 26 1179279AI826538, AL038664, AI051237, AI267318, AI688542, AI052104, AW 163010, AI376453, AW192514, AI818589, AW029328, AI678648, AI566340, AI972077, AI811155, AI936746, AI089502, AI372947, AI004230, AI354532, AL119666, AW074007, AI084362, AI027083, AI691080, AA862706, AA621070, AI744332, AI149953, AW242075, AI149949, AI150745, AI199180, AI625208, AI003733, W20002, AI627187, AW 130451, AW270647, AI014764, AI091649, AA041468, W55944, AW149580, AI445868, AW151070, AA148318, AI092273, AI005484, AI372493, AA040575, AA595861, AI689545, AI524423, AI521587, AA342697, AA908191, AI689268, AI270577, AI952557, AI372494, AI619883, AI538583, AW263138, AI026832, AA040673, W25901, AI368864, AW316596, AI539834, AA349447, AA721376, AW292131, 819495, AA662403, AW085967, T75472, AA808860, AA199620, AI125767, AW090571, H20652, N78681, AA176087, N32970, AI074758, AA740389, AA814692, AA300365, AA329440, AI547225, AW275741, AA894651, AA302328, AI680268, T48533, AA719848, F13229, AA383093, AA386145, AA970611, AI536066, D31244, 244196, H20558, AI350433, AW243606, AI784415, AI676163, AA063203, D82747, W26208, AA471277, AA903068, AA664940, AA853050, AA897635, AW021288, D31438, AI535982, AI419708, AI680414, AA090164, AA362084, AA386197, 862151, AI250661, 862259, W28043, AI521566, AI611841, F10830, AA355685, AA343846, 843842, AA334321, T69962, AA093703, T24661, AA845417, D56184, AA332748, 240172, 839290, T27330, AI695489, D80027, 812857, 838429, AI524545, AA095572, W 15187, T28780, F24108, AA176086, AW37.5368, AA323934, AA337850, D81428, AA373966, AA344329, AW293901, AA039822, AW375337, T69912, AI802643, F35697, AA848160, AI523217, AI241878, AA148317, H60591, AI709179, T25879, AA970902, N63253, T79010, AW 169467, T16724, AA093662, N84238, AW241154, AW270806, AI198549, AW380896, AI630928, AL038665, AI284509, AI557186, AL041150, AI433157, AI554821, AW 151136, AI539771, AI537677, AI494201, AI500659, AI815232, AI801325, AI500523, AI436429, AI582932, AI923989, AI284517, AI371228, AI500706, AI445237, AI491776, AW 151138, AI889189, AI521560, AI500662, AI889168, AI866573, AI633493, AI434256, AI805769, AI888661, AI284513, AI888118, AI889147, AI440252, AI610557, AI366900, AI538850, AI872423, AW 172723, AI440263, AI866469, AI434242, AI859991, AI866786, AI860003, AI242736, AI887499, AL080007, AW 151132, AL045500, AI866465, D31885, AE000658, U85195, AF223953, AF172088, AF133669, I89947, AL049938, AL122101, A58524, A58523, AF113019, AL133072, AL122049, AR011880, AL050138, U35846, AF111851, AF017437, AL049423, Y09972, AL133113, A77033, A77035, AL110280, AL050277, AL137459, AF124728, E06743, AR038854, AL137476, AL133640, L31396, L31397, AL137539, AL110221, AL122110, Y16645, AL133080, I33392, AL133077, AB029066, AL137550, E07361, AF061943, AC002471, AC005374, AF097996, A65340, AL050024, Y11587, AL133081, AF185614, AL137463, A21625, U91329, AF076464, AF113677, U67958, U58996, U42766, X72889, U30290, AF081195, X00474, AF028823, AL137526, AL117583, AL117585, AL122123, I09360, A18777, AL049300, AL117460, AF158248, AL049283, X84990, AL133075, U49434, AL133565, S78214, I00734, AF078844, AF113694, AL133084, AF118094, AL133070, AL110197, AL133098, AL049466, E00617, E00717, E00778, AL133015, A03736, AJ012755, AL110269, S61953, AL137298, E03671, I42402, I46765, AL117457, AL096744, AL050108, AF102578, AF104032, 566283, AL049447, I89944, AB031064, L19437, AL117649, Y10823, 249216, X61399, AR034821, AF100931, AF113691, AL133558, A45787, AL133608, U78525, AL110222, X63574, AF008439, AW273036, AW473757, AW474360, AW510479, AW514205, AW516993, AW518951, AW630807, AW662059, and AW 664721. .

18 HKZCK47 28 1045503H95452, N30257, N40659, N41980, W32409, W32855, AA001776, AA747202, AA876069, D81852, D81929, D81992, AI373734, AI299574, AI221043, AI910470, and AW 196676.

21 HRDEP41 31 1129342AI819066, AI096889, N57566, AI652475, AI762701, AI571414, AW072021, AW136982, AA770246, AI360284, AI185794, AA653289, AI632543, N39261, AI093864, AI361124, W78162, AW072658, AA434490, AI092771, W79205, AL044476, AI126467, AI377502, AI371895, AA086103, AI768304, N50074, AI347630, W57608, AI004516, AI684524, AA702096, AI992245, AA434395, AI360808, AI359558, AA324953, AA781993, AI933385, AA026053, AI243341, AI654422, H09521, AA086102, AA789226, AA988193, H09520, 245285, W28501, 240993, C04672, AI623520, AA969690, AI674810, AW138990, AI214707, AI368537, N46707, 249982, U30172, and 249983.

22 HKGBJ74 32 1189265AW245735, AW245623, AW245881, N57252, AW409766, AW247020, AW250562, AI885802, AW250222, AI829136, H98166, AW072012, AW409650, AA236737, AA130880, AA081326, AA176211, AA081347, AA618083, AA176210, AA486434, AI092870, AI589540, AW273066, AI745511, AI953980, AW409675, AA524975, AW 173680, AW 149138, AW248461, AA056429, T66277, AI686881, AW246286, AA633178, AI361334, AA313766, AA081346, AI972465, AA878148, AA828127, AA135151, AA490323, AA541297, AI208260, AA448202, N31322, AI283002, AW 102925, AA132698, AA024484, AA814452, AI278141, AI338188, AI802624, AA922736, N35048, AI002040, AA502359, AW079983, AA634343, AA564581, AW251043, AI143056, N21370, AI087009, AL045365, AI038842, AA081325, AA235062, AA724588, AA234910, AW272966, AA219233, AW245379, AA219293, AA380625, AA135288, AA147048, AW002314, AA627088, 244936, T66190, AA604261, AA490427, AA132809, AW263614, AA845407, AI300044, AA393562, AA939182, AA309709, AA936143, AA298671, AA298672, T90842, 802758, AA579652, AA025328, AA297935, AL135190, AI798307, AA054656, AA296918, F 12298, AI024248, AI480173, N45060, AW250136, AI952399, AA297613, 802757, AA298705, AI469323, AA130935, AW247374, AI766398, AA876843, AA297025, AA346949, AW380104, AA551740, AA285283, AA309007, AI337489, AA090542, T19272, AI799323, T73385, F09925, AA090691, AA598486, AI627371, AA102203, AA147239, AW246489, AI358227, AW380065, AW409756, AA759368, AI954095, AI434969, AI241741, AW 198112, AW 176261, AI612913, AI554821, AI680221, AI868204, AI613038, AI580436, AI345688, AI289791, AI961589, AI698391, AI440238, AI582932, AI583578, AL120254, AI669639, AI565031, AI537187, AL039086, AI973272, AW129929, AI433157, AI702073, AI633125, AI432570; AW 172982, AI539260, AI690389; AL042745, AI345415, AW084151, AI610362, AA937574, AI687568, AI43361 I, AI590043, AI309306, AI584130, AI251221, AW022682, AW163834, AI638644, AI927233, AI559863, AI624938, AI699823, AL042515, AL037582, AI799183, AL037602, AI582966, AI583065, AI801793, AW 167146, AW 167926, _ AL040241, AI249389, AI161279, N75779, AL036954, AI475339, AI889189, AA464646, AL046466, AI623835, AI471909, AI628325, AI590134, 869938, AI524654, AI318280, AI886415, AA954134; AA595932, AL118781, AW 149069, AI933992, AI491710, AI114703, AI621341, AI254042, AW268080, AW075669, AI916419, AW089387, AI811840, AA693331, AI565172, AI345612, AW022636, AI685517, AL134830, AW020397, AI690748, AW 150557, AI635016, AI679959, AL043981, AI954080, AW025279, AI524179, AI446511, AI280637, AF090385, AF161489, AF046025, AF110956, AB024303, AL137530, U77594, I89947, A65340, AF113699, AL133619, I33392, I09499, M92439, X99226, AL049324, AL117416, AL050138, AL137292, AR034821, X83508, I48978, AL137558, S68736, AB007812, AL136884, S36676, AL133665, I32738, AF200464, AL049938, A65341, AF079763,.AL137533, U73682, AL117435, A03736, AL133080, AF176651, AF102578, 297214, E04233, AF038847, 237987, AF177401, AF044323, AL096720, AL137550, U35846, E01614, E13364, AL080110, A08913, AL137480, AL080124, I26207, AF026124, A08912, AF139986, A08910, AF106862, AR020905, A08909, AL117587, AR038854, E02914, A08908, AF159615, AF090900, S77771, A12297, AL133049, S76508, S78453, AL049283, AL137459, AL110221, AF113676, AL137641, AF090934, AF183393, AF090903, AL110222, D83032, AF017437, X80340, S53987, X63574, I00734, S61953, A18777, AL122110, AF118070, AF107847, AF069506, U58996, AJ242859, X52128, AF017790, AF 106697, AL 133016, E00617, E00717, E00778, A21101, U72621, AF061573, AL137476, A15345, I89931, AF028823, AL049339, A83556, A77033, A77035, AL117460, AF117657, AL080140, AJ003118, AL133568, AL117394, AL122093, AL050092, AL096751, AF032666, I49625, AL137271, U86379, AF000301, AL137529, AF090901, X65873, AL 122098, AF 1 I
1112, A08907, AF100781, AL133558, A65336, D44497, I52013, AF047716, Y09972, AL137548, AL050366, A08911, I29004, I89934, X98066, AL133067, 282022, AL122100, AL137711, AL080126, AL110280, X63410,.583456;

AL133104, AF126247, I48979, AF141289, 582852, AB025103, U42031, AL080127, AL117578, M27260, AL110225, AL110224, X93495, AJ005690, X53587, Y10655, AL133637, AF118094, Y11254, AL080159, AR054987, U68233, I92592, X92070, M96857, AL023657, AF055917, AF185576, AF137367, AF008439, I68732, AF013214, AF017152, AF207750, AF113690, L13297, A07588, X66871, AF067728, AL133640, AF111849, AF125948, AF199027, AL122121, AF030513, AL137665, AL080163, AF057300, AF057299, AJ012755, S75997, AL137526, X99257, AF111851, U95114, 213966, E01812, AF047443, U35146, AL137538, AL133557, AL050149, U57352, D16301, AF081197, AF081195, X72889, S78214, AF113691.;

AF026816, AL137463, AL117648, X82434, _ ' AF113677, AL050277, A86558, AF090943, AL122050, AL137523, AL049382, AL049314, AF115392, X63162, AF153205, AJ238278, AL117585, AL080234, U96683, S69510, AL133075, U72620, A08916, AR011880, AF115410, and AW467171.

22 HKGBJ74 67 1025650AW245735, AW245623, AW245881, N57252, AW247020, AW250562, AI885802, AW250222, AI829136, H98166, AW072012, AW409650, AA236737, AW409766, AA130880, AA081326, AA176211, AA081347, AA618083, AA 176210, AA486434, AI092870, AI589540, AW273066, AI745511, AI953980, AW409675, AA5249'7S, AW 173680, AW 149138, AW248461, T66277, AI686881, AW246286, AA633178, AAOS6429, AI361334, AA313766, AA081346, AI972465, AA878148, AA828127, AA135151;

AA490323, AA541297, AI208260, AA448202, N31322, AI283002, AW102925, AA132698, AA024484, AA8144S2, AI278141, AI338188, AI802624, AA922736, N35048, AI002040, AA502359, AW079983, AA634343, AA564581, AW251043, AI143056, N21370, AI038842, AA081325, AI087009, AA235062, AA724588, AA234910, AW272966, AA219233, AW245379, AA219293, AA380625, AA135288, AA147048, AW002314, AA627088, 244936, T66190, AL04S365, AA490427, AA132809, AW263614, AA845407, AI300044, AA393562, AA939182, AA309709, AA936143, AA298671, AA298672, T90842, 802758, AA604261, AA579652, AA025328, AA297935, AL135190, AI798307, AA054656, AA296918, F12298, N45060, AI024248, AA297613, 802757, AA298705, AI9S2399, AI469323, AA130935, AI480173, AW247374, AI766398, AA876843, AA297025, AA346949, AW250136, AW380104, AA551740, AA285283, AA309007, AI337489, AA090542, T19272, AI799323, T73385, F09925, AA090691, AA598486, AI627371, AA102203, AW246489, AA147239, AI358227, AW380065, AA759368, AW409756, AI339364, AF090385, AF161489, AF046025, AF110956, AB024303, and AW467171.

Having generally described the invention, the same will be more readily understood by reference to the following examples, which are provided by way of illustration and are not intended as limiting.
Examples Example 1: Isolation of a Selected cDl~lA Clone From the Deposited Sample Each cDNA clone in a cited ATCC deposit is contained, in a plasmid vector.
Table 1 identifies the vectors used to construct the cDNA library from which-each clone was isolated. In many cases, the vector used to construct the library is a phage vector from which a plasmid has been excised. The table immediately below correlates the related plasmid for each phage vector used in constructing the cDNA
library. For example, where a particular clone is identified in Table 1 as being isolated in the vector "Lambda Zap," the corresponding deposited clone is in "pBluescript."
Vector Used to Construct Library Corresponding Deposited Plasmid Lambda Zap pBluescript (pBS) Uni-Zap XR pBluescript (pBS) Zap Express pBK

lafinid BA plafmid BA

pSportl pSportl pCMVSport 2.0 pCMVSport 2.0 pCMVSport 3.0 pCMVSport 3.0 pCR~2.1 pCR2.1 Vectors Lambda Zap (U.S. Patent Nos. 5,128,256 and 5,286,636), Uni-Zap XR (U.S. Patent Nos. 5,128, 256 and 5,286,636), Zap Express (U.S. Patent Nos.
5,128,256 and 5,286,636), pBluescript (pBS) (Short, J. M. et al., Nucleic Acids Res.
16:7583-7600 (1988); Alting-Mees, M. A. and Short, J. M., Nucleic Acids Res.
17:9494 (1989)) and pBK (Aping-Mees, M. A. et al., Strategies 5:58-61 (1992)) are commercially available from Stratagene Cloning Systems, Inc., 11011 N. Torrey Pines Road, La Jolla, CA, 92037. pBS contains an ampicillin resistance gene and -, pBK contains a neomycin resistance gene. Both can be transformed into E. coli strain XL-1 Blue, also available from Stratagene. pBS comes in 4 dorms SK+, SK-, KS+
and KS. The S and K refers to the orientation of the polylinker to the T7 and primer sequences which flank the polylinker region ("S" is for SacI and "K" is for KpnI which are the first sites on each respective end of the linker). "+" or "-" refer to the orientation of the fl origin of replication ("ori"), such that in one orientation, single stranded rescue initiated from the fl on generates sense strand DNA and in the other, antisense.
Vectors pSportl, pCMVSport 2.0 and pCMVSport 3.0, were obtained from Life Technologies, Inc., P. O. Box 6009, Gaithersburg, MD 20897. All Sport vectors contain an ampicillin resistance gene and may be transformed into E. coli strain DH10B, also available from Life Technologies. (See, for instance, Gruber, C.
E., et al., Focus 15:59 (1993).) Vector lafmid BA (Bento Soares, Columbia University, NY) contains an ampicillin resistance gene and can be transformed into E. coli strain XL-1 Blue. Vector pCR~2.1, which is available from Invitrogen, 1600 Faraday Avenue, Carlsbad, CA 92008, contains an ampicillin resistance gene and may be transformed into E. coli strain DH10B, available from Life Technologies. (See, for instance, Clark, J. M., Nuc. Acids Res. 16:9677-9686 (1988) and Mead, D. et al., Bio/Technology 9: (1991).) Preferably, a polynucleotide of the present invention does not comprise the phage vector sequences identified for the particular clone in Table 1, as well as the corresponding plasmid vector sequences designated above.
The deposited material in the sample assigned the ATCC Deposit Number cited in Table 1 for any given cDNA clone also may contain one or more additional plasmids, each comprising a cDNA clone different from that given clone. Thus, deposits sharing the same ATCC Deposit Number contain at least a plasmid for each cDNA clone identified in Table 1. Typically, each ATCC deposit sample cited in Table 1 comprises a mixture of approximately equal amounts (by weight) of about 50 plasmid DNAs, each containing a different cDNA clone; but such a deposit sample may include plasmids for more or less than 50 cDNA clones, up to about 500 cDNA
clones.
Two approaches can be used to isolate a particular clone from the deposited sample of plasmid DNAs cited for that clone in Table 1. First, a plasmid is directly -.

isolated by screening the clones using a polynucleotide probe corresponding to SEQ
ID NO:X.
Particularly, a specific polynucleotide with 30-40 nucleotides is synthesized using an Applied Biosystems DNA synthesizer according to the sequence reported.
The oligonucleotide is labeled, for instance, with 32P-y-ATP using T4 polynucleotide kinase and purified according to routine methods. (E.g., Maniatis et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring, NY
(1982).) The plasmid mixture is transformed into a suitable host, as indicated above (such as XL-1 Blue (Stratagene)) using techniques known to those of skill in the art, such as those provided by the vector supplier or in related publications or patents cited above.
The transformants are plated on 1.5% agar plates (containing the appropriate selection agent, e.g., ampicillin) to a density of about 150 transformants (colonies) per plate.
These plates are screened using Nylon membranes according to routine methods for bacterial colony screening (e.g., Sambrook et al., Molecular Cloning: A
Laboratory Manual, 2nd Edit., (1989), Cold Spring Harbor Laboratory Press, pages 1.93 to 1.104), or other techniques known to those of skill in the art.
Alternatively, two primers of 17-20 nucleotides derived from both ends of the SEQ ID NO:X (i.e., within the region of SEQ ID NO:X bounded~by the 5' NT and the 3' NT of the clone defined in Table 1) are synthesized and used to amplify the desired cDNA using the deposited cDNA plasmid as a template. The polymerase chain reaction is carried out under routine conditions, for instance, in 25 u1 of reaction mixture with 0.5 ug of the above cDNA template. A convenient reaction mixture is 1.5-5 mM MgClz, 0.01 % (w/v) gelatin, 20 uM each of dATP, dCTP, dGTP, dTTP, 25 pmol of each primer and 0.25 Unit of Taq polymerase. Thirty five cycles of PCR
(denaturation at 94 degree C for 1 min; annealing at 55 degree C for 1 min;
elongation at 72 degree C for 1 min) are performed with a Perkirt-Elmer Cetus automated thermal cycler. The amplified product is analyzed by agarose gel electrophoresis and the DNA band with expected molecular weight is excised and purified. The PCR
product is verified to be the selected sequence by subcloning and sequencing the DNA product.
Several methods are available for the identification of the 5' or 3' non-coding portions of a gene which may not be present in the deposited clone. These methods include but are not limited to, filter probing, clone enrichment using specific probes, and protocols similar or identical to 5' and 3' "RACE" protocols which are well known in the art. For instance, a method similar to 5' RACE is available for generating the missing 5' end of a desired full-length transcript. (Fromont-Racine et al., Nucleic Acids Res. 21(7):1683-1684 (1993).) Briefly, a specific RNA oligonucleotide is ligated to the 5' ends of a population of RNA presumably containing full-length gene RNA transcripts. A
primer set containing a primer specific to the ligated RNA oligonucleotide and a primer specific to a known sequence of the gene of interest is used to PCR
amplify the 5' portion of the desired full-length gene. This amplified product may then be sequenced and used to generate the full length gene.
This above method starts with total RNA isolated from the desired source, although poly-A+ RNA can be used. The RNA preparation, can then be treated with phosphatase if necessary to eliminate 5' phosphate groups on degraded or damaged RNA which may interfere with the later RNA lipase step. The phosphatase should then be inactivated and the RNA treated with tobacco acid pyrophosphatase in order to remove the cap structure present at the 5' ends of messenger RNAs. This reaction leaves a 5' phosphate group at the S' end of the cap cleaved RNA which can then be ligated to an RNA oligonucleotide using T4 RNA lipase.
This modified RNA preparation is used as a template for first strand cDNA
synthesis using a gene specific oligonucleotide. The first strand synthesis reaction is used as a template for PCR amplification of the desired 5' end using a primer specific to the ligated RNA oligonucleotide and a primer specific to the known sequence of the gene of interest. The resultant product is then sequenced and analyzed to confirm that the 5' end sequence belongs to the desired gene.
Example 2: Isolation of Genomic Clones Corresponding to a Polynucleotide A human genomic P1 library (Genomic Systems, Inc.) is screened by PCR
using primers selected for the cDNA sequence corresponding to SEQ 1D NO:X., according to the method described in Example 1. (See also, Sambrook.) Example 3: Tissue Distribution of Polypeptide Tissue distribution of mRNA expression of polynucleotides of the present invention is determined using protocols for Northern blot analysis, described by, among others, Sambrook et al. For example, a cDNA probe produced by the method described in Example 1 is labeled with P32 using the rediprimeTM DNA labeling system (Amersham Life. Science), according to manufacturer's instructions.
After labeling, the probe is purified using CHROMA SPIN-100TM column (Clontech Laboratories, Inc.), according to manufacturer's protocol number PT1200-1. The purified labeled probe is then used to examine various human tissues for mRNA
expression.
Multiple Tissue Northern (MTN) blots containing various human tissues (H) or human immune system tissues (IM) (Clontech) are examined with the labeled probe using ExpressHybTM hybridization solution (Clontech) according to manufacturer's protocol number PT1190-1. Following hybridization and washing, the blots are mounted and exposed to film at -70 degree C overnight, and the films developed according to standard procedures.
Example 4: Chromosomal Mapping of the Polynucleotides An oligonucleotide primer set is designed according to the sequence at the 5' end of SEQ ID NO:X. This primer preferably spans about 100 nucleotides. This primer set is then used in a polymerase chain reaction under the following set of conditions : 30 seconds,95 degree C; 1 minute, 56 degree C; 1 minute, 70 degree C.
This cycle is repeated 32 times followed by one 5 minute cycle at 70 degree C.
Human, mouse, and hamster DNA is used as template in addition to a somatic cell hybrid panel containing individual chromosomes or chromosome fragments (Bios, Inc). The reactions is analyzed on either 8% polyacrylamide gels or 3.5 %
agarose gels. Chromosome mapping is determined by the presence of an approximately 100 by PCR fragment in the particular somatic cell hybrid.
Example 5: Bacterial Expression of a Polypeptide A polynucleotide encoding a polypeptide of the present invention is amplified using PCR oligonucleotide primers corresponding to the 5' and 3' ends of the DNA

sequence, as outlined in Example 1, to synthesize insertion fragments. The primers used to amplify the cDNA insert should preferably contain restriction sites, such as BamHI and XbaI, at the 5' end of the primers in order to clone the amplified product into the expression vector. For example, BamHI and XbaI correspond to the restriction enzyme sites on the bacterial expression vector pQE-9. (Qiagen, Inc., Chatsworth, CA). This plasmid vector encodes antibiotic resistance (Amps, a bacterial origin of replication (ori), an IPTG-regulatable promoter/operator (P/0), a ribosome binding site (RBS), a 6-histidine tag (6-His), and restriction enzyme cloning sites.
The pQE-9 vector is digested with BamHI and XbaI and the amplified fragment is ligated into the pQE-9 vector maintaining the reading frame initiated at the bacterial RBS. The ligation mixture is then used to transform the E. coli strain M15/rep4 (Qiagen, Inc.) which contains multiple copies of the plasmid pREP4, which expresses the lacI repressor and also confers kanamycin resistance (Kan~.
Transformants are identified by their ability to grow on LB plates and ampicillin/kanamycin resistant colonies are selected. Plasmid DNA is isolated and confirmed by restriction analysis.
Clones containing the desired constructs are grown overnight (0/N) in liquid culture in LB media supplemented with both Amp (100 ug/ml) and Kan (25 ug/ml).
The O/N culture is used to inoculate a large culture at a ratio of 1:100 to 1:250. The cells are grown to an optical density 600 (O.D.boo) of between 0.4 and 0.6.
IPTG
(Isopropyl-B-D-thiogalacto pyranoside) is then added to a final concentration of 1 mM. IPTG induces by inactivating the lacI repressor, clearing the P/O leading to increased gene expression.
Cells are grown for an extra 3 to 4 hours. Cells are then harvested by centrifugation (20 mins at 6000Xg). The cell pellet is solubilized in the chaotropic agent 6 Molar Guanidine HCl by stirring for 3-4 hours at 4 degree C. The cell debris is removed by centrifugation, and the supernatant containing the polypeptide is loaded onto a nickel-nitrilo-tri-acetic acid ("Ni-NTA") affinity resin column (available from QIAGEN, Inc., supra). Proteins with a 6 x His tag bind to the Ni-NTA resin with high affinity and can be purified in a simple one-step procedure (for details see: The QIAexpressionist (1995) QIAGEN, Inca, supra).

Briefly, the supernatant is loaded onto the column in 6 M guanidine-HC1, pH
8, the column is first washed with 10 volumes of 6 M~ guanidine-HCI, pH 8, then washed with 10 volumes of 6 M guanidine-HCl pH 6, and finally the polypeptide is . eluted with 6 M guanidine-HCI, pH 5.
The purified protein is then renatured by dialyzing it against phosphate-buffered saline (PBS) or 50 mM Na-acetate, pH 6 buffer plus 200 mM NaCI.
Alternatively, the protein can be successfully refolded while immobilized on the Ni-NTA column. The recommended conditions are as follows: renature using a linear 6M-1M urea gradient in 500 mM NaCI, 20% glycerol, 20 mM Tris/HCl pH 7.4, containing protease inhibitors. The renaturation should be performed over a period of 1.5 hours or more. After renaturation the proteins are eluted by the addition of 250 mM immidazole. Immidazole is removed by a final dialyzing step against PBS or mM sodium acetate pH 6 buffer plus 200 mM NaCI. The purified protein is stored at 4 degree C or frozen at -80 degree C.
In addition to the above expression vector, the present invention further includes an expression vector comprising phage operator and promoter elements operatively linked to a polynucleotide of the present invention, called pHE4a.. (ATCC
Accession Number 209645, deposited on February 25, 1998.) This vector contains:
1 ) a neomycinphosphotransferase gene as a selection marker, 2) an E. coli origin of replication, 3) a T5 phage promoter sequence, 4) two lac operator sequences, 5) a Shine-Delgarno sequence, and 6) the lactose operon repressor gene (lacIq). The origin of replication (oriC) is derived from pUCl9 (LTI, Gaithersburg, MD).
The promoter sequence and operator sequences are made synthetically. , DNA can be inserted into the pHEa by restricting the vector with NdeI and XbaI, BamHI, XhoI, or Asp718, running the restricted product on a gel, and isolating the larger fragment (the stuffer fragment should be about 310 base pairs). The DNA
insert is generated according to the PCR protocol described in Example l, using PCR
primers having restriction sites for NdeI (5' primer) and XbaI, BamHI, XhoI, or Asp718 (3' primer). The PCR insert is gel purified and restricted with compatible enzymes. The insert and vector are ligated according to standard protocols.
The engineered vector could easily be substituted in the above protocol to express protein in a bacterial system.

Example 6: Purification of a Polypeptide from an Inclusion Body The following alternative method can be used to purify a polypeptide expressed in E coli when it is present in the form of inclusion bodies. Unless otherwise specified, all of the following steps are conducted at 4-10 degree C.
Upon completion of the production phase of the E. coli fermentation, the cell culture is cooled to 4-10 degree C and the cells harvested by continuous centrifugation at 15,000 rpm (Heraeus Sepatech). On the basis of the expected yield of protein per unit weight of cell paste and the amount of purified protein required, an appropriate amount of cell paste, by weight, is suspended in a buffer solution containing 100 mM Tris, 50 mM EDTA, pH 7.4. The cells are dispersed to a homogeneous suspension using a high shear mixer.
The cells are then lysed by passing the solution through a microfluidizer (Microfuidics, Corp. or APV Gaulin, Inc.) twice at 4000-6000 psi. The homogenate 15- is then mixed with NaCI solution to a final concentration of 0.5 M NaCI, followed by centrifugation at 7000 xg for 15 min. The resultant pellet is washed again using O.SM
NaCI, 100 mM Tris, 50 mM EDTA, pH 7.4.
The resulting washed inclusion bodies are solubilized with 1.5 M guanidine hydrochloride (GuHCI) for 2-4 hours. After 7000 xg centrifugation for 1 S
min., the pellet is discarded and the polypeptide containing supernatant is incubated at 4 degree C overnight to allow further GuHCI extraction. .
Following high speed centrifugation (30,000 xg) to remove insoluble particles, the GuHCI solubilized protein is refolded by quickly mixing the GuHCI extract with 20 volumes of buffer containing 50 mM sodium, pH 4.5, 150 mM NaCI; 2 mM EDTA
by vigorous stirnng. The refolded diluted protein solution is kept at 4 degree C
without mixing for 12 hours prior to further purification steps.
To clarify the refolded polypeptide solution, a previously prepared tangential filtration unit equipped with 0.16 um membrane filter with appropriate surface area (e.g., Filtron), equilibrated with 40 mM sodium acetate, pH 6.0 is employed.
The filtered sample is loaded onto a canon exchange resin (e.g., Poros HS-50, Perseptive Biosystems). The column is washed with 40 mM sodium acetate, pH 6.0 and eluted with 250 mM, 500 mM, 1000 mM, and 1500 mM NaCI in the same buffer, in a stepwise manner. The absorbance at 280 nm of the effluent is continuously monitored. Fractions are collected and further analyzed by SDS-PAGE.
Fractions containing the polypeptide are then pooled and mixed with 4 volumes of water. The diluted sample is then loaded onto a previously prepared set of tandem columns of strong anion (Poros HQ-50, Perseptive Biosystems) and weak anion (Poros CM-20, Perseptive Biosystems) exchange resins. The columns are equilibrated with 40 mM sodium acetate, pH 6Ø Both columns are washed with mM sodium acetate, pH 6.0, 200 mM NaCI. The CM-20 column is then eluted using a 10 column volume linear gradient, ranging from 0.2 M NaCI, 50 mM sodium acetate, pH 6.0 to 1.0 M NaCI, 50 mM sodium acetate, pH 6.5. Fractions are , collected under constant AZ$o monitoring of the effluent. Fractions containing the polypeptide (determined, for instance, by 16% SDS-PAGE) are then pooled.
The resultant polypeptide should exhibit greater than 95% purity after the above refolding and purification steps. No major contaminant bands should be observed from Commassie blue stained 16% SDS-PAGE gel when 5 ug of purified protein is loaded. The purified protein can also be tested for endotoxin/LPS
contamination, and typically the LPS content is less than 0.1 ng/ml according to LAL
assays. .
Example 7: Cloning and Expression of a Polvnentide in a Baculovirus Expression System In this example, the plasmid shuttle vector pA2 is used to insert a polynucleotide into a baculovirus to express a polypeptide. This expression vector contains the strong polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus (AcMNPV) followed by convenient restriction sites such as BamHI, Xba I and Asp718.~ The polyadenylation site of the simian virus 40 ("SV40") is used for efficient polyadenylation. For easy selection of recombinant virus, the plasmid contains the beta-galactosidase gene from E. coli under control of a weak Drosophila promoter in the same orientation, followed by the polyadenylation signal of the polyhedrin gene. The inserted genes are flanked on both sides by viral sequences for cell-mediated homologous recombination with wild-type: viral DNA
to generate a viable virus that express the cloned polynucleotide.

Many other baculovirus vectors can be used in place of the vector above, such as pAc373, pVL941, and pAcIMI, as one skilled in the art would readily appreciate, as long as the construct provides appropriately located signals for transcription, translation, secretion and the like, including a signal peptide and an in-frame AUG as required. Such vectors are described, for instance, in Luckow et al., Virology 170:31 39 (1989).
Specifically, the cDNA sequence contained in the deposited clone, including the AUG initiation codon and the naturally associated leader sequence identified in Table 1, is amplified using the PCR protocol described in Example 1. If the naturally ' occurnng signal sequence is used to produce the secreted protein, the pA2 vector does not need a second signal peptide. Alternatively, the vector can be modified (pA2 GP) to include a baculovirus leader sequence, using the standard methods described in Summers et al., "A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures," Texas Agricultural Experimental Station Bulletin No. 1555 (1987).
The amplified fragment is isolated from a 1 % agarose gel using a commercially available kit ("Geneclean," BIO 101 Inc., La Jolla, Ca.). The fragment then is digested with appropriate restriction.enzymes and again purified on a 1%
agarose gel.
The plasmid is digested with the corresponding restriction enzymes and optionally, can be dephosphorylated using calf intestinal phosphatase, using routine procedures known in the art. The DNA is then isolated from a 1 % agarose gel using a commercially available kit ("Geneclean" BIO 101 Inc., La Jolla, Ca.).
The fragment and the dephosphorylated plasmid are ligated together with T4 DNA ligase. E. coli HB101 or other suitable E. coli hosts such as XL-1 Blue (Stratagene Cloning Systems, La Jolla, CA) cells are transformed with the ligation mixture and spread on culture plates. Bacteria containing the plasmid are identified by digesting DNA from individual colonies and analyzing the digestion product by gel electrophoresis. The sequence of the cloned fragment is confirmed by DNA
sequencing.
Five ug of a plasmid containing the polynucleotide is co-transfected with 1.0 ug of a commercially available linearized baculovirus DNA ("BaculoGoldTM

baculovirus DNA", Pharmingen, San Diego, CA), using the lipofection method described by Felgner et al., Proc. Natl. Acad. Sci. USA 84:7413-7417 (1987).
One ug of BaculoGoldTM virus DNA and 5 ug of the plasmid are mixed in a sterile well of a microtiter plate containing 50 u1 of serum-free Grace's medium (Life Technologies Inc., Gaithersburg, MD). Afterwards, 10 u1 Lipofectin plus 90 u1 Grace's medium are added, mixed and incubated for 15 minutes at room temperature. Then the transfection mixture is added drop-wise to S~ insect cells (ATCC CRL 1711) seeded in a 35 mm tissue culture plate with 1 ml Grace's medium without serum. The plate is then incubated for 5 hours at 27 degrees C. The transfection solution is then removed from the plate and 1 ml of Grace's insect medium supplemented with 10% fetal calf serum is added. Cultivation is then continued at 27 degrees C for four days.
After four days the supernatant is collected and a plaque assay is performed, as described by Summers and Smith, supra. An agarose gel with "Blue Gal" (Life Technologies Inc., Gaithersburg) is used to allow easy identification and isolation of gal-expressing clones, which produce blue-stained plaques. (A detailed description of a "plaque assay" of this type can also be found in the user's guide for insect cell culture and baculovirology distributed by Life Technologies Inc., Gaithersburg, page 9-10.) After appropriate incubation, blue stained plaques are picked with the tip of a micropipettor (e.g., Eppendorf). The agar containing the recombinant viruses is then resuspended in a microcentrifuge tube containing 200 u1 of Grace's medium and the suspension containing the recombinant baculovirus is used to infect S~ cells seeded in 35 mm dishes. Four days later the supernatants of these culture dishes are harvested and then they are stored at 4 degree C.
To verify the expression of the polypeptide, Sf~ cells are grown in Grace's medium supplemented with 10% heat-inactivated FBS. The cells are infected with the recombinant baculovirus containing the polynucleotide at a multiplicity of infection ("MOI") of about 2. If radiolabeled proteins are desired, 6 hours later the medium is removed and is replaced with SF900 II medium minus methionine and cysteine (available from Life Technologies Inc., Rockville, MD). A$er 42 hours, 5 uCi of 35S-methionine and 5 uCi 35S-cysteine (available from Amersham) are added.
The cells are further incubated for 16 hours and then are harvested by centrifugation.

The proteins in the supernatant as well as the intracellular proteins are analyzed by SDS-PAGE followed by autoradiogi-aphy (if radiolabeled).
Microsequencing of the amino acid sequence of the amino terminus of purified protein may be used to determine the amino terminal sequence of the produced protein.
Example 8: Expression of a Polyueutide in Mammalian Cells The polypeptide of the present invention can be expressed in a mammalian cell. A typical mammalian expression vector contains a promoter element, which .
mediates the initiation of transcription of mRNA, a protein coding sequence, and signals required for the termination of transcription and polyadenylation of the transcript. Additional elements include enhancers, Kozak sequences and intervening sequences flanked by donor and acceptor sites for RNA splicing. Highly efficient transcription is achieved with the early and late promoters from SV40, the long terminal repeats (LTRs) from Retroviruses, e.g., RSV, HTLVI, HIVI and the early promoter of the cytomegalovirus (CMV). However, cellular elements can also be used (e.g., the human actin promoter).
Suitable expression vectors for use, in practicing the present invention include, for example, vectors such as pSVL and pMSG (Pharmacia, Uppsala, Sweden), pRSVcat (ATCC 37152), pSV2dhfr,(ATCC 37146), pBCI2MI (ATCC 67109), pCMVSport 2.0, and pCMVSport 3Ø Mammalian host cells that could be used include, human Hela, 293, H9 and Jurkat cells, mouse NIH3T3 and C127 cells, Cos 1, Cos 7 and CV1, quail QC1-3 cells, mouse L cells and Chinese hamster ovary (CHO) cells.
Alternatively, the polypeptide can be expressed in stable cell lines containing the polynucleotide integrated into a chromosome. The co-transfection with a selectable marker such as dhfr, gpt, neomycin, hygromycin allows the identification and isolation of the transfected cells.
The transfected gene can also be amplified to express large amounts-of the encoded protein. The DHFR (dihydrofolate reductase) marker is useful in developing cell lines that carry several hundred or even several thousand copies of the gene of interest. (See, e.g., Alt, F. W., et al., J. Biol. Chem. 253:1357-1370 (1978);
Hamlin, J. _, L. and Ma, C., Biochem. et Biophys. Acta, 1097:107-143 (1990); Page, M. J. and Sydenham, M. A., Biotechnology 9:64-68 (1991).) Another useful selection marker /is the enzyme glutamine synthase (GS) (Murphy et al., Biochem J. 227:277-279 (1991); Bebbington et al., Bio/Technology 10:169=175 (1992). Using these markers, the mammalian cells are grown in selective medium and the cells with the highest resistance are selected. These cell lines contain the amplified genes) integrated into a chromosome. Chinese hamster ovary (CHO) and NSO cells are often used for the production of proteins.
Derivatives of the plasmid pSV2-dhfr (ATCC Accession No. 37146), the expression vectors pC4 (ATCC Accession No. 209646) and pC6 (ATCC Accession No.209647) contain the strong promoter (LTR) of the Rous Sarcoma Virus (Cullen et al., Molecular and Cellular Biology; 438-447 (March, 1985)) plus a fragment of the CMV-enhancer (Boshart et al., Cell 41:521-530 (1985).) Multiple cloning sites, e.g., with the restriction enzyme cleavage sites BamHI, XbaI and Asp718, facilitate the cloning of the gene of interest. The vectors also contain the 3' intron, the polyadenylation and termination signal of the rat preproinsulin gene, and the mouse DHFR gene under control of the SV40 early promoter.
. Specifically, the plasmid pC6, for example, is digested with appropriate restriction enzymes and then dephosphorylated using calf intestinal phosphates by procedures known in the art. The vector is then isolated from a 1 % agarose gel.
A polynucleotide of the present invention is amplified according to the protocol outlined in Example 1. If the naturally occurring signal sequence is used to produce the secreted protein, the vector does not need a second signal peptide.
Alternatively, if the naturally occurring signal sequence is not used, the vector can be modified to include a heterologous signal sequence. (See, e.g., WO 96/34891.) The amplified fragment is isolated from a 1 % agarose gel using a commercially available kit ("Geneclean," BIO 101 Inc., La Jolla, Ca.). The fragment then is digested with appropriate restriction enzymes and again purified on a agarose gel.
The amplified fragment is then digested with the same restriction enzyme and purified on a 1 % agarose gel. The isolated fragment and the dephosphorylated vector are then ligated with T4 DNA ligase. E. coli HB101 or XL-1 Blue cells are then _.

transformed and bacteria are identified that contain the fragment inserted into plasmid pC6 using, for instance, restriction enzyme analysis.
Chinese hamster ovary cells lacking an active DHFR gene is used for transfection. Five pg of the expression plasmid pC6 a pC4 is cotransfected with 0.5 ug of the plasmid pSVneo using lipofectin (Felgner et al., supra). The plasmid pSV2-neo contains a dominant selectable marker, the neo gene from Tn5 encoding an enzyme that confers resistance to a group of antibiotics including 6418. The cells are seeded in alpha minus MEM supplemented with 1 mg/ml 6418. After 2 days, the cells are trypsinized and seeded in hybridoma cloning plates (Greiner, Germany) in alpha minus MEM supplemented with 10, 25, or 50 nglml of metothrexate plus 1 mg/ml 6418. After about 10-14 days single clones are trypsinized and then seeded in 6-well petri dishes or 10 ml flasks using different concentrations of methotrexate (50 nM, 100 nM, 200 nM, 400 nM, 800 nM). Clones growing at the highest concentrations of methotrexate are then transferred to new 6-well plates containing even higher concentrations of methotrexate (1 uM, 2 uM, 5 uM, 10 mM, 20 mM).
The same procedure is repeated until clones are obtained which grow at a concentration of 100 - 200 uM. Expression of the desired gene product is analyzed, for instance, by SDS-PAGE and Western blot or by reversed phase HPLC analysis.
Example 9: Protein Fusions The polypeptides of the present invention are preferably fused to other proteins. These fusion proteins can be used for a variety of applications. For example, fusion of the present polypeptides to His-tag, HA-tag, protein A, IgG
domains, and maltose binding protein facilitates purification. (See Example 5;
see also EP A 394,827; Traunecker, et al., Nature 331:84-86 (1988).) Similarly, fusion to IgG-1, IgG-3, and albumin increases the halflife time in vivo. Nuclear localization signals fused to the polypeptides of the present invention can target the protein to a specific subcellular localization, while covalent heterodimer or homodimers can increase or decrease the activity of a fusion protein. Fusion proteins can also create chimeric molecules having more than one function. Finally, fusion proteins can increase solubility and/or stability of the fused protein compared to the non-fused protein. All of the types of fusion proteins described above can be made by modifying the following protocol, which outlines the fusion of a polypeptide to an IgG molecule, or the protocol described in Example 5 Briefly, the human Fc portion of the IgG molecule can be PCR amplified, using primers that span the 5' and 3' ends of the sequence described below.
These primers also should have convenient restriction enzyme sites that will facilitate cloning into an expression vector, preferably a mammalian expression vector.
For example, if pC4 (Accession No. 209646) is used, the human Fc portion can be ligated into the BamHI cloning site. Note that the 3' BamHI site should be destroyed. Next, the vector containing the human Fc portion is re-restricted with BamHI, linearizing the vector, and a polynucleotide of the present invention, isolated by the PCR protocol described in Example 1, is ligated into this BamHI site.
Note that the polynucleotide is cloned without a stop codon, otherwise a fusion protein will not be produced.
If the naturally occurnng signal sequence is used to produce the secreted protein, pC4 does not need a second signal peptide. Alternatively, if the naturally occurring signal sequence is not used, the vector can be modified to include a heterologous signal sequence. (See, e.g., WO 96/34891 Human IgG Fc region:
GGGATCCGGAGCCCAAATCTTCTGACAAAACTCACACATGCCCACC
GTGCCCAGCACCTGAATTCGAGGGTGCACCGTCAGTCTTCCTCTTCCCCCC
AAA.ACCCAAGGACACCCTCATGATCTCCCGGACTCCTGAGGTCACATGCG
TGGTGGTGGACGTAAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTAC
GTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGC
AGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAG
GACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCT
CCCAACCCCCATCGAGAA.AACCATCTCCAA.AGCCAAAGGGCAGCCCCGA
GAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAA
CCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCAAGCGACATCG
CCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCAC
GCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCAC
CGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGA

TGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCT
CCGGGTAAATGAGTGCGACGGCCGCGACTCTAGAGGAT (SEQ )17 NO:1) Example 10: Production of an Antibody from a Polypeptide The antibodies of the present invention can be prepared by a variety of methods. (See, Current Protocols, Chapter 2.) As one example of such methods, cells expressing a polypeptide of the present invention is administered to an animal to induce the production of sera containing polyclonal antibodies. In a preferred method, a preparation of the secreted protein is prepared and purified to render it substantially free of natural contaminants. Such a preparation is then introduced into an animal in order to produce polyclonal antisera of greater specific activity.
In the most preferred method, the antibodies of the present invention are monoclonal antibodies (or protein binding fragments thereof). Such monoclonal antibodies can be prepared using hybridoma technology. (Kohler et al., Nature 256:495 (1975); Kohler et al., Eur. J. Immunol. 6:511 (1976); Kohler et al., Eur. J. Immunol. 6:292 (1976); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas, Elsevier, N.Y., pp. 563-681 (1981).) In general, such procedures involve immunizing an animal (preferably a mouse) with polypeptide or, more preferably, with a secreted polypeptide-expressing cell. Such cells may be cultured in any suitable tissue culture medium; however, it is preferable to culture cells in Earle's modified Eagle's medium supplemented with 10% fetal bovine serum (inactivated at about 56 degrees C), and supplemented with about 10 g/1 of nonessential amino acids, about 1,000 U/ml of penicillin, and about 100 ug/ml of streptomycin.
The splenocytes of such mice are extracted and fused with a suitable myeloma cell line. Any suitable myeloma cell line may be employed in accordance with the present invention; however, it is preferable to employ the parent myeloma cell line (SP20), available from the ATCC. After fusion, the resulting hybridoma cells are selectively maintained in HAT medium, and then cloned by limiting dilution as described by Wands et al. (Gastroenterology 80:225-232 (1981).) The hybridoma .
cells obtained through such a selection are then assayed to identify clones which secrete antibodies capable of binding the polypeptide.

Alternatively, additional antibodies capable of binding to the polypeptide can be produced in a two-step procedure using anti-idiotypic antibodies. Such a method makes use of the fact that antibodies are themselves antigens, and therefore, it is possible to obtain an antibody which binds to a second antibody. In accordance with this method, protein specific antibodies are used to immunize an animal, preferably a mouse. The splenocytes of such an animal are then used to produce hybridoma cells, and the hybridoma cells are screened to identify clones which produce an antibody whose ability to bind to the protein-specific antibody can be blocked by the polypeptide. Such antibodies comprise anti-idiotypic antibodies to the protein=
specific antibody and can be used to immunize an animal to induce formation of further protein-specific antibodies.
It will be appreciated that Fab and F(ab')2 and other fragments of the antibodies of the present invention may be used according to the methods disclosed herein. Such fragments are typically produced by proteolytic cleavage, using enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab')2 fragments). Alternatively, secreted protein-binding fragments can be produced through the application of recombinant DNA technology or through synthetic chemistry.
For in vivo use of antibodies in humans, it may be preferable to use "humanized" chimeric monoclonal antibodies. Such antibodies can be produced using genetic constructs derived from hybridoma cells producing the monoclonal antibodies described above. Methods for producing chimeric antibodies are known in the art. (See, for review, Morrison, Science 229:1202 (1985); Oi et al., BioTechniques 4:214 (1986); Cabilly et al., U.S. Patent No. 4,816,567;
Taniguchi et al., EP 171496; Morrison et al., EP 173494; Neuberger et al., WO 8601533;
Robinson et al., WO 8702671; Boulianne et al., Nature 312:643 (1984); Neuberger et al., Nature 314:268 (1985).) Example 11: Production Of Secreted Protein For High-Throughput Screening As_ says The following protocol produces a supernatant containing a polypeptide to be tested. This supernatant can then be used in the Screening Assays described herein. _.

First, dilute Poly-D-Lysine (644 587 Boehringer-Mannheim) stock solution ( lmg/ml in PBS) 1:20 in PBS (w/o calcium or magnesium 17-516F Biowhittaker) for .
a working solution of SOug/ml. Add 200 u1 of this solution to each well (24 well plates) and incubate at RT for 20 minutes. Be sure to distribute the solution over each . well (note: a 12-channel pipetter may be used with tips on every other channel).
Aspirate off the Poly-D-Lysine solution and rinse with lml PBS (Phosphate Buffered Saline). The PBS should remain in the well until just prior to plating the cells and plates may be poly-lysine coated in advance for up to two weeks.
Plate 293T cells (do not carry cells past P+20) at 2 x 105 cells/well in :5m1 DMEM(Dulbecco's Modified Eagle Medium)(with 4.5 G/L glucose and L-glutamine (12-604F Biowhittaker))/10% heat inactivated FBS(14-503F Biowhittaker)/lx Penstrep(17-602E Biowhittaker). Let the cells grow overnight.
The next day, mix together in a sterile solution basin: 300 u1 Lipofectamine (18324-012' Gibco/BRL) and Sml Optimem I (31985070 Gibco/BRL)/96-well plate.
With a small volume mufti-channel pipetter, aliquot approximately tug of an expression vector containing a polynucleotide insert, produced by the methods described in Examples 8 or 9, into an appropriately labeled 96-well round bottom plate. With a mufti-channel pipetter, add SOuI of the Lipofectamine/Optimem I
mixture to each well. Pipette up and down gently to mix. Incubate at RT 15-45 minutes. After about 20 minutes, use a mufti-channel pipetter to add 1 SOuI
Optimem I to each well. As a control, one plate of vector DNA lacking an insert should be transfected with each set of transfections.
Preferably, the transfection should be performed by tag-teaming the following tasks. By tag-teaming, hands on time is cut in half, and the cells do not spend too much time on PBS. First, person A aspirates off the media.from four 24-well-plates of cells, and then person B rinses each well with .5-lml PBS. Person A then aspirates off PBS rinse, and person B, using alt-channel pip.etter with tips on every other channel, adds the 200u1 of DNA/Lipofectamine/Optimem I complex to the odd wells first, then to the even wells, to each row on the 24-well plates. Incubate at 37 degrees C for 6 hours.
While cells are incubating, prepare appropriate media, either 1 %BSA in DMEM with lx penstrep, or CHO-5 media (116.6 mg/L of CaCl2 (anhyd); 0.00130 mg/L CuS04-5H20; 0.050 mg/L of Fe(N03)3-9Hz0; 0.417 mg/L of FeS04-7H20;
311.80 mg/L of Kcl; 28.64 mg/L of MgClz; 48.84 mg/L of MgS04; 6995.50 mg/L of NaCI; 2400.0 mg/L of NaHC03; 62.50 mg/L of NaHZPOa-HzO; 71.02 mg/L of NazHP04; .4320 mg/L of ZnS04-7H20; .002 mg/L of Arachidonic Acid ; 1.022 mg/L
of Cholesterol; .070 mg/L of DL-alpha-Tocopherol-Acetate; 0.0520 mg/L of Linoleic Acid; 0.010 mg/L of Linolenic Acid; 0.010 mg/L of Myristic Acid; 0.010 mg/L of Oleic Acid; 0.010 mg/L of Palmitric Acid; 0.010 mg/L of Palinitic Acid; 100 mg/L of Pluronic F-68; 0.010 mg/L of Stearic Acid; 2.20 mg/L of Tween 80; 4551 mg/L of D-Glucose; 130.85 mg/ml of L- Alanine; 147.50 mg/ml of L-Arginine-HCL; 7.50 mglml of L-Asparagine-HzO; 6.65 mg/ml of L-Aspartic Acid; 29.56 mg/ml of L-Cystine-2HCL-HzO; 31.29 mg/ml of L-Cystine-2HCL; 7.35 mg/ml of L-Glutamic Acid; 365.0 mg/ml of L-Glutamine; 18.75 mg/ml of Glycine; 52.48 mg/ml.of L-Histidine-HCL-HzO; 106.97 mg/ml of L-Isoleucine; 111.45 mg/ml of L-Leucine; 163.75 mg/ml of L-Lysine HCL; 32.34 mg/ml of L-Methionine; 68.48 mg/ml of L-Phenylalainine; 40.0 mg/ml of L-Proline; 26.25 mg/ml of L-Serine; 101.05 mg/ml of L-Threonine;
19.22 mg/ml of L-Tryptophan; 91.79 mg/ml of L-Tryrosine-2Na-2Hz0; 99.65 mg/ml of L-Valine; 0.0035 mg/L of Biotin; 3.24 mg/L of D-Ca Pantothenate; 11.78 mg/L of Choline Chloride; 4.65 mg/L of Folic Acid; 15.60 mg/L of i-Inositol; 3.02 mg/L
of Niacinamide; 3.00 mg/L of Pyridoxal HCL; 0.031 mg/L of Pyridoxine HCL; 0.319 mg/L of Riboflavin; 3.17 mg/L of Thiamine HCL; 0.365 mg/L of Thymidine; and 0.680 mg/L of Vitamin Biz; 25 mM of HEPES Buffer; 2.39 mg/L of Na Hypoxanthine; 0.105 mg/L of Lipoic Acid; 0.081 mg/L of Sodium Putrescine-2HCL;
55.0 mg/L of Sodium Pyruvate; 0.0067 mg/L of Sodium Selenite; 20uM of Ethanolamine; 0.122 mg/L-of Ferric Citrate; 41.70 mglL of Methyl-B-Cyclodextrin complexed with Linoleic Acid; 33.33 mg/L of Methyl-B-Cyclodextrin complexed with Oleic Acid; and 10 mg/L of Methyl-B-Cyclodextrin complexed with Retinal) with 2mm glutamine and 1 x penstrep. (BSA (81-068-3 Bayer) 100gm dissolved in DMEM for a 10% BSA stock solution). Filter the media and collect 50 u1 for endotoxin assay in 15m1 polystyrene conical.
The transfection reaction is terminated, preferably by tag-teaming, at the end of the incubation period. Person A aspirates off the transfection media, while person B adds 1.5m1 appropriate media to each well. Incubate at 37 degrees C for 45 or 72 hours depending on the media used: 1 %BSA for 45 hours or CHO-5 for 72 hours.
On day four, using a 300u1 multichannel pipetter, aliquot 600u1 in one lml deep well plate and the remaining supernatant into a 2m1 deep well. The supernatants from each well can then be used in the assays described in Examples 13-20.
It is specifically understood that when activity is obtained in any of the assays described below using a supernatant, the activity originates from either the polypeptide directly (e.g., as a secreted protein) or by the polypeptide inducing expression of other proteins, which are then secreted into the supernatant.
Thus, the invention further provides a method of identifying the protein in the supernatant characterized by an activity in a particular assay.
Example 12: Construction of GAS Reporter Construct One signal transduction pathway involved in the differentiation and proliferation of cells is called the Jaks-STATs pathway. Activated proteins in the Jaks-STATs pathway bind to gamma activation site "GAS" elements or interferon-sensitive responsive element ("ISRE"), located in the promoter of many genes.
The binding of a protein to these elements alter the expression of the associated gene.
GAS and ISRE elements are recognized by a class of transcription factors called Signal Transducers and Activators of Transcription, or "STATs." There are six members of the STATs family. Statl and Stat3 are present in many cell types, as is Stat2 (as response to IFN-alpha is widespread). Stat4 is more restricted and is not in many cell types though it has. been found in T helper class I, cells after treatment with IL-12. StatS was originally called mammary growth factor, but has been found at higher concentrations in other cells including myeloid cells. It can be activated in tissue culture cells by many cytokines.
The STATs are activated to translocate from the cytoplasm to the nucleus upon tyrosine phosphorylation by a set of kinases known as the Janus Kinase ("Jaks") family. Jaks represent a distinct family of soluble tyrosine kinases and include Tyk2, Jakl, Jak2, and Jak3. These kinases display significant sequence similarity and are generally catalytically inactive in resting cells.

The Jaks are activated by a wide range of receptors summarized in the Table below. (Adapted from review by Schidler and Darnell, Ann. Rev. Biochem. 64:621-51 (1995).) A cytokine receptor family, capable of activating Jaks, is divided into two groups: (a) Class 1 includes receptors for IL-2, IL-3, IL-4, IL-6, IL-7, IL-9, IL-11, IL-12, IL-15, Epo, PRL, GH, G-CSF, GM-CSF, LIF, CNTF, and thrombopoietin; and (b) ' Class 2 includes IFN-a, IFN-g, and IL-10. The Class 1 receptors share a conserved cysteine motif (a set of four conserved cysteines and one tryptophan) and a WSXWS
motif (a membrane proximal region encoding Trp-Ser-Xxx-Trp-Ser (SEQ ID N0:2)).
Thus, on binding of a ligand to a receptor, Jaks are activated, which in turn activate STATs, which then translocate and bind to GAS elements. This entire process is encompassed in the Jaks-STATs signal transduction pathway.
Therefore, activation of the Jaks-STATs pathway, reflected by the binding of the GAS or the ISRE element, can be used to indicate proteins involved in the proliferation and differentiation of cells. For example, growth factors and cytokines are known to activate the Jaks-STATs pathway. (See Table below.) Thus, by using GAS elements linked to reporter molecules, activators of the Jaks-STATs pathway can be identified.

JAKs STATS GAS(elements) or ISRE

LiQand t1~1C2Jakl JaIQ Jak3 IFN family IFN-a/B + + - - 1,2,3 ISRE

IFN-g + + - 1 GAS (IRF 1 >Lys6>IFP) Il-10 + ? ? - 1,3 gp130 family -IL-6 (Pleiotrophic)+ + + ? 1,3 GAS (IRF1>Lys6>IFP).

Il-11(Pleiotrophic)? + ? ? 1,3 OnM(Pleiotrophic) ? + + ? 1,3 LIF(Pleiotrophic)? + + ? 1,3 CNTF(Pleiotrophic) -/+ + + ? 1,3 G-CSF(Pleiotrophic)? + ? ? 1,3 IL-12(Pleiotrophic)+ _ + + 1,3 Q-C family IL-2 (lymphocytes) - + - + 1,3,5GAS

IL-4 (lymph/myeloid)- + - + 6 GAS (IRF1 = IFP Ly6)(IgH)-IL-7 (lymphocytes) - + - + 5 GAS

IL-9 (lymphocytes) - + - + 5 GAS

IL-13 (lymphocyte) - + ? ? 6 GAS

IL-15 ? + ? + 5 GAS

gp 140 family IL-3 (myeloid) - - + - 5 GAS (IRF1>IFPLy6) IL-5 (myeloid) - - + - S GAS

GM-CSF (myeloid) - - + - 5 GAS

Growth hormone family GH ? - + - 5 PRL ? +/- + - 1,3,5 EPO ? - + - 5 GAS(B-CAS>IRF1=IFPLy6) Receptor Tyrosine Kinases EGF ? + + - 1,3 GAS (IRF1) PDGF ? + + . 1,3 -CSF-1 ? + + - 1,3 GAS (not IRF1) To construct a synthetic GAS containing promoter element, which is used in the Biological Assays described in Examples 13-14, a PCR based strategy is employed to generate a GAS-SV40 promoter sequence. The 5' primer contains four tandem copies of the GAS binding site found in the IRF1 promoter and previously demonstrated to bind STATs upon induction with a range of cytokines (Rothman et al., Immunity l :457-468 (1994).), although other GAS or ISRE elements can be used instead. The 5' primer also contains l8bp of sequence complementary to the early promoter sequence and is flanked with an XhoI site. The sequence of the 5' primer is:
5':GCGCCTCGAGATTTCCCCGAAATCTAGATTTCCCCGAAATGATTT
CCCCGAAATGATTTCCCCGAAATATCTGCCATCTCAATTAG:3' (SEQ ID
N0:3) The downstream primer is complementary to the SV40 promoter and is flanked with a Hind III site: 5':GCGGCAAGCTTTTTGCAAAGCCTAGGC:3' (SEQ ID N0:4) PCR amplification is performed using the SV40 promoter template present in the B-gal:promoter plasmid obtained from Clontech. The resulting PCR fragment is digested with XhoI/Hind III and subcloned into BLSK2-. (Stratagene.) Sequencing with forward and reverse primers confirms that the insert contains the following sequence:
5':CTCGAGATTTCCCCGAAATCTAGATTTCCCCGAAATGATTTCCCC
GAAATGATTTCCCCGAAATATCTGCCATCTCAATTAGTCAGCAACCATAGT
CCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCA
TTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGC
CGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAG
GCCTAGGCTTTTGCAAAAAGCTT:3' (SEQ ID N0:5) With this GAS promoter element linked to the SV40 promoter, a GAS:SEAP2 reporter construct is next engineered. Here, the reporter molecule is a secreted alkaline phosphatase, or "SEAP." Clearly, however, any reporter molecule can be instead of SEAP, in this or in any of the other Examples. Well known reporter molecules that can be used instead of SEAP include chloramphenicol acetyltransferase (CAT), luciferase, alkaline phosphatase, B-galactosidase, green fluorescent protein (GFP), or any protein detectable by an antibody.
The above sequence confirmed synthetic GAS-SV40 promoter element is subcloned into the pSEAP-Promoter vector obtained from Clontech using HindIII
and XhoI, effectively replacing the SV40 promoter with the amplified GAS:SV40 promoter element, to create the GAS-SEAP vector. However, this vector does not contain a neomycin resistance gene, and therefore, is not preferred for mammalian expression systems.
Thus, in order to generate mammalian stable cell lines expressing the GAS-SEAP reporter, the GAS-SEAP cassette is removed from the GAS-SEAP vector using SaII and NotI, and inserted into a backbone vector containing the neomycin resistance gene, such as pGFP-1 (Clontech), using these restriction sites in the multiple cloning site, to create the GAS-SEAP/Neo vector. Once this vector is transfected into .
mammalian cells, this vector can then be used as a reporter molecule for GAS
binding as described in Examples 13-14.
Other constructs can be made using the above description and replacing GAS
with a different promoter sequence. For example, construction of reporter molecules containing NFK-B and EGR promoter sequences are described in Examples 15 and 16. However, many other promoters can be substituted using the protocols described in these Examples. For instance, SRE, IL-2, NEAT, or Osteocalcin promoters can be substituted, alone or in combination (e.g., GAS/NF-KB/EGR, GAS/NF-KB, Il-2/NFAT, or NF-KB/GAS). Similarly, other cell lines can be used to test reporter construct activity, such as HELA (epithelial), HUVEC (endothelial), Reh (B-cell), Saos-2 (osteoblast), HUVAC (aortic), or Cardiomyocyte.
Example 13: High-Throu~hnut Screening Assav for T-cell Activi The following protocol is used to assess T-cell activity by identifying factors, and determining whether supernate containing a polypeptide of the invention proliferates and/or differentiates T-cells. T-cell activity is assessed using the GAS/SEAP/Neo construct produced in Example 12. Thus, factors that increase SEAP
activity indicate the ability to activate the Jaks-STATS signal transduction pathway.
The T-cell used in this assay is Jurkat T-cells (ATCC Accession No. TIB-152), although Molt-3 cells (ATCC Accession No. CRL-1552) and Molt-4 cells (ATCC
Accession No. CRL-1582) cells can also be used.
Jurkat T-cells are lymphoblastic CD4+ Thl helper cells. Iri order to generate stable cell lines, approximately 2 million Jurkat cells are transfected with the GAS-SEAP/neo vector using DMRIE-C (Life Technologies)(transfection procedure described below). The transfected cells are seeded to a density of approximately 20,000 cells per well and transfectants resistant to 1 mg/ml genticin selected.
Resistant colonies are expanded and then tested for their response to increasing concentrations of interferon gamma. The dose response of a selected clone is demonstrated.
Specifically, the following protocol will yield sufficient cells for 75 wells containing 200 u1 of cells. Thus, it is either scaled up, or performed in multiple to generate sufficient cells for multiple 96 well plates. Jurkat cells are maintained in RPMI + 10% serum with.1 %Pen-Strep. Combine 2.5 mls of OPTI-MEM (Life Technologies) with 10 ug of plasmid DNA in a T25 flask. Add 2.5 ml OPTI-MEM
containing 50 u1 of DMRIE-C and incubate at room temperature for 15-45 mins.
During the incubation period, count cell concentration, spin down the required number of cells (10' per transfection), and resuspend in OPTI-MEM to a final concentration of 10' cells/ml. Then add lml of 1 x 10' cells in OPTI-MEM to flask and incubate at 37 degrees C for 6 hrs. After the incubation, add 10 ml of RPMI
+ 15% serum.
The Jurkat:GAS-SEAP stable reporter lines are maintained in RPMI + 10%
serum, 1 mg/ml Genticin, and 1 % Pen-Strep. These cells are treated with supernatants containing polypeptides of the invention and/or induced polypeptides of the invention as produced by the protocol described in Example 11.
On the day of treatment with the supernatant, the cells should be washed and resuspended in fresh RPMI + 10% serum fo a density of 500,000 cells per ml.
The exact number of cells required will depend on the number of supernatants being screened. For one 96 well plate, approximately 10 million cells (for 10 plates, 100 million cells) are required.

Transfer the cells to a triangular reservoir boat, in order to dispense the cells into a 96 well dish, using a 12 channel pipette. Using a 12 channel pipette;
transfer 200 u1 of cells into each well (therefore adding 100, 000 cells per well).
After all the plates have been seeded, 50 u1 of the supernatants are transferred S directly from the 96 well plate containing the supernatants into each well using a 12 channel pipette. In addition, a dose of exogenous interferon gamma (0.1, 1.0,

10 ng) is added to wells H9, H10, and H11 to serve as additional positive controls for the assay.
The 96 well dishes containing Jurkat cells treated with supernatants are placed in an incubator for 48 hrs (note: this time is variable between 48-72 hrs). 35 u1 samples from each well are then transferred to an opaque 96 well plate using a channel pipette. The opaque plates should be covered (using sellophene covers) and stored at -20 degrees C until SEAP assays are performed according to Example 17.
The plates containing the remaining treated cells are placed at 4 degrees C
and serve as a source of material for repeating the assay on a specific well if desired.
As a positive control, 100 Unit/ml interferon gamma can be used which is known to activate Jurkat T cells. Over 30 fold induction is typically observed in the positive control wells.
The above protocol may be used in the generation of both transient, as well as, stable transfected cells, which would be apparent to those of skill in the art.
Example 14: High-Throughput ScreeninyAssay Identifying Myeloid Activity The following protocol is used to assess myeloid activity by determining whether polypeptides of the invention proliferates and/or differentiates myeloid cells.
Myeloid cell activity is assessed using the GAS/SEAP/Neo construct produced in Example 12. Thus, factors that increase SEAP activity indicate the ability to activate the Jaks-STATS signal transduction pathway. The myeloid cell used in this assay is U937, a pre-monocyte cell line, although TF-1, HL60, or KG1 can be used.
To transiently transfect U937 cells with the GAS/SEAP/Neo construct produced in Example 12, a DEAE-Dextran method (Kharbanda et. al., 1994, Cell Growth & Differentiation, 5:259-265) is used. First, harvest 2x 10e7 U937 cells and-wash with PBS. The U937 cells are usually grown in RPMI 1640 medium containing -10% heat-inactivated fetal bovine serum (FBS) supplemented with 100 units/ml penicillin and 100 mg/ml streptomycin.
Next, suspend the cells in 1 ml of 20 mM Tris-HCl (pH 7.4) buffer containing 0.5 mg/ml DEAF-Dextran, 8 ug GAS-SEAP2 plasmid DNA, 140 mM NaCI, 5 mM
KCI, 375 uM Na2HP04.7H20, 1 mM MgCl2, and 675 uM CaCl2. Incubate at 37 degrees C for 45 min.
Wash the cells with RPMI 1640 medium containing 10% FBS and then resuspend in 10 ml complete medium and incubate at 37 degrees C for 36 hr.
The GAS-SEAP/LT937 stable cells are obtained by growing the cells in 400 ug/ml 6418. The 6418-free medium is used for routine growth but every one to two months, the cells should be re-grown in 400 ug/ml 6418 for couple of passages.
These cells are tested byharvesting 1x10$ cells (this is enough for ten 96-well plates assay) and wash with PBS. Suspend the cells in 200 ml above described growth medium, with a final density of 5x105 cells/ml. Plate 200 u1 cells per well in the 96-well plate (or 1x105 cells/well):
Add 50 u1 of the supernatant prepared by the protocol described in Example

11. Incubate at 37 degrees C for 48 to 72 hr., As a positive control, 100 UnitJml interferon gamma can be used which is known to activate U937 cells. Over 30 fold induction is typically observed in the positive control wells. SEAP assay the supernatant according to the protocol described in Example 17.
Example 15: High-Throughput Screening Assay Identifying Neuronal Activity.
When cells undergo differentiation and proliferation, a group of genes are activated through many, different signal transduction pathways. One of these genes, EGR1 (early growth response gene 1), is induced in various tissues and cell types upon activation. The promoter of EGR1 is responsible for such induction. Using the EGR1 promoter linked to reporter molecules, activation of cells can be assessed.
Particularly, the following protocol is used to assess neuronal activity in cell lines. PC 12 cells (rat phenochromocytoma cells) are known to proliferate and/or differentiate by activation with a number of mitogens, such as TPA
(tetradecanoyl phorbol acetate), NGF (nerve growth factor), and EGF (epidermal growth factor).
The EGR1 gene expression is activated during this treatment. Thus, by stably transfecting PC 12 cells with a construct containing an EGR promoter linked to SEAP
reporter, activation of PC12 cells can be assessed.
The EGR/SEAP reporter construct can be assembled by the following protocol. The EGR-1 promoter sequence (-633 to +1)(Sakamoto K et al., Oncogene S 6:867-871 (1991)) can be PCR amplified from human genomic DNA using the following primers:
5' GCGCTCGAGGGATGACAGCGATAGAACCCCGG -3' (SEQ >D N0:6) 5' GCGAAGCTTCGCGACTCCCCGGATCCGCCTC-3' (SEQ )D N0:7) Using the GAS:SEAP/Neo vector produced in Example 12, EGRl amplified product can then be inserted into this vector. Linearize the GAS:SEAP/Neo vector using restriction enzymes XhoI/HindIII, removing the GAS/SV40 stuffer.
Restrict the EGR1 amplified product with these same enzymes. Ligate the vector and the EGR1 promoter.
To prepare 96 well-plates for cell culture, two mls of a coating solution (1:30 dilution of collagen type I (Upstate Biotech Inc. Cat#08-115) in 30% ethanol (filter sterilized)) is added per one 10 cm plate or 50 ml per,well of the 96-well plate, and allowed to air dry for 2 hr.
PC12 cells are routinely grown in RPMI-1640 medium (Bio Whittaker) containing 10% horse serum (JRH BIOSCIENCES, Cat. # 12449-78P), 5% heat-inactivated fetal bovine serum (FBS) supplemented with 100 units/ml penicillin and 100 ug/ml streptomycin on a precoated 10 cm tissue culture dish. One to four split is done every three to four days. Cells are removed from the plates by scraping and resuspended with pipetting up and down for more than 15 times.
Transfect the EGR/SEAP/Neo construct into PC 12 using the Lipofectamine protocol described in Example 11. EGR-SEAP/PC12 stable cells are obtained by growing the cells in 300 ug/ml 6418. The 6418-free medium is used for routine growth but every one~to two months, the cells should be re-grown in 300 ug/ml for couple of passages.
To assay for neuronal activity, a 10 cm plate with cells around 70 to 80%
confluent is screened by removing the old medium. Wash the cells once with PBS
(Phosphate buffered saline). Then starve the cells in low serum medium (RPMI-containing 1 % horse serum and 0.5% FBS with antibiotics) overnight. _ The next morning, remove the medium and wash the cells with PBS. Scrape off the cells from the plate, suspend the cells well in 2 ml low serum medium.
Count the cell number and add more low serum medium to reach final cell density as Sx 1 OS
cells/ml.
Add 200 u1 of the cell suspension to each well of 96-well plate (equivalent to 1x105 cells/well). Add SO u1 supernatant produced by Example 11, 37oC for 48 to 72 hr. As a positive control, a growth factor known to activate PC12 cells through EGR
can be used, such as 50 ng/ul of Neuronal Growth Factor (NGF). Over fifty-fold induction of SEAP is typically seen in the positive control wells. SEAP assay the supernatant according to Example 17.
Example 16: High-Throughput Screening Assay for T-cell Activity NF-KB (Nuclear Factor KB) is a transcription factor activated by a wide variety of agents including the inflammatory cytokines IL-1 and TNF, CD30 and CD40, lymphotoxin-alpha and lymphotoxin-beta, by exposure to LPS or thrombin, and by expression of certain viral gene products. As a transcription factor, NF-KB
regulates the expression of genes involved in immune cell activation, control of apoptosis (NF- KB appears to shield cells from apoptosis), B and T-cell development, anti-viral and antimicrobial responses, and multiple stress responses.
In non-stimulated conditions, NF- KB is retained in the cytoplasm with I-KB
(Inhibitor KB). However, upon stimulation, I- KB is phosphorylated and degraded, causing NF- KB to shuttle to the nucleus, thereby activating transcription of target genes. Target genes activated by NF- KB include IL-2, IL-6, GM-CSF, ICAM=1 and class 1 MHC.
Due to its central role and ability to respond to a range of stimuli, reporter constructs utilizing the NF-KB promoter element are used to screen the supernatants produced in Example 11. Activators or inhibitors of NF-KB would be useful in treating diseases. For example, inhibitors of NF-KB could be used to treat those diseases related to the acute or chronic activation of NF-KB, such as rheumatoid 30. arthritis.
To construct a vector containing the NF-KB promoter element, a PCR based strategy is employed. The upstream primer contains four tandem copies of the NF-KB binding site (GGGGACTTTCCC) (SEQ ID N0:8), 18 by of sequence complementary to the 5' end of the SV40 early promoter sequence, and is flanked with an XhoI site:
5':GCGGCCTCGAGGGGACTTTCCCGGGGACTTTCCGGGGACTTTCC
GGGACTTTCCATCCTGCCATCTCAATTAG:3' (SEQ ID N0:9) The downstream primer is complementary to the 3' end of.the SV40 promoter and is flanked with a Hind III site:
S':GCGGCAAGCTTTTTGCAAAGCCTAGGC:3' (SEQ ID N0:4) PCR amplification is performed using the SV40 promoter template present in the pB-gal:promoter plasmid obtained from Clontech. The resulting PCR fragment is digested with XhoI and Hind III and subcloned into BLSK2-. (Stratagene) Sequencing with the T7 and T3 primers confirms the insert contains the following sequence:.
~ 5':CTCGAGGGGACTTTCCCGGGGACTTTCCGGGGACTTTCCGGGAC
TTTCCATCTGCCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTC
CGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATG
GCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTG
AGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGC
AAAA.AGCTT:3' (SEQ ID NO:10) Next, replace the SV40 minimal promoter element present in the pSEAP2-promoter plasmid (Clontech) with this NF-KB/SV40 fragment using XhoI and HindIII. However, this vector does not contain a neomycin resistance gene, and therefore, is not preferred for mammalian expression systems.
In order to generate stable mammalian cell lines, the NF-KB/SV40/SEAP
cassette is removed from the above NF-KB/SEAP vector using restriction enzymes SaII and NotI, and inserted into a vector containing neomycin resistance.
Particularly, the NF-KB/SV40/SEAP cassette was inserted into pGFP-1 (Clontech), replacing the GFP gene, after restricting pGFP-1 with SaII and NotI.
Once NF-KB/SV40/SEAP/Neo vector is created, stable Jurkat T-cells are created and maintained according to the protocol described in Example 13.
Similarly, _ the method for assaying supernatants with these stable Jurkat T-cells is also described in Example ,13. As a positive control, exogenous TNF alpha (0.1, l, 10 ng) is added to wells H9, H10, and H11, with a 5-10 fold activation typically observed.
Example 17: Assay for SEAP Activity As a reporter molecule for the assays described in Examples 13-16, SEAP
activity is assayed using the Tropix Phospho-light Kit (Cat. BP-400) according to the following general procedure. The Tropix Phospho-light Kit supplies the Dilution, Assay, and Reaction Buffers used below.
Prime a dispenser with the 2.5x Dilution Buffer and dispense 15 u1 of 2.5x dilution buffer into Optiplates containing 35 u1 of a supernatant. Seal the plates with a plastic sealer and incubate at 65 degree C for 30 min. Separate the Optiplates to avoid uneven heating.
Cool the samples to room temperature for 15 minutes. Empty the dispenser and prime with the Assay Buffer. Add 50 ml Assay Buffer and incubate at room temperature 5 min. Empty the dispenser and prime with the Reaction Buffer (see the table below). Add 50 u1 Reaction Buffer and incubate at room temperature for minutes. Since the intensity of the chemiluminescent signal is time dependent, and it takes about 10 minutes to read 5 plates on luminometer, one should treat 5 plates at each time and start the second set 10 minutes later.
Read the relative light unit in the luminometer. Set H12 as blank, and print the results. An increase in chemiluminescence indicates reporter activity.
Reaction Buffer Formulation:
# of Rxn buffer diluent CSPD
fates (ml) (ml) 11 65 3.25

12 . 70 3.5

13 . ' 75 3.75 ~

14 80 4

15 - 85 4.25

16 90 4.5

17 95 4.75

18 100 S

19 105 5.25

20 110 5.5 . 21 115 5.75 23 125 6.25 24 130 6.5 25 135 6.75 27 145 7.25 28 150 7.5 29 155 7.75 31 165 8.25 32 170 8.5 33 175 8.75 35 . 185 9.25 36 190 9.5 37 195 9.75 39 205 ~ 10.25 40 210 10.5 41 215 10.75 43 225 11.25 44 230 11.5 45 235 11.75 47 245 12.25 48 250 12.5 49 255 12.75 Example 18: High-Throughput Screening Assay Identifyin~ Changes in Small -Molecule Concentration and Membrane Permeability Binding of a ligand to a receptor is known to alter intracellular levels of small molecules, such as calcium, potassium, sodium, and pH, as well as alter membrane potential. These alterations can be measured in an assay to identify supernatants which bind to receptors of a particular cell. Although the following protocol describes an assay for calcium, this protocol can easily be modified to detect changes in potassium, sodium, pH, membrane potential, or any other small molecule which is detectable by a fluorescent probe.

The following assay uses Fluorometric Imaging Plate Reader ("FLIPR") to measure changes in fluorescent molecules (Molecular Probes) that bind small molecules. Clearly, any fluorescent molecule detecting a small molecule can be used instead of the calcium fluorescent molecule, fluo-4 (Molecular Probes, Inc.;
catalog no. F-14202), used here.
For adherent cells, seed the cells at 10,000 -20,000 cells/well in a Co-star black 96-well plate with clear bottom. The plate is incubated in a COZ
incubator for 20 hours. The adherent cells are washed two times in Biotek washer with 200 u1 of HBSS (Hank's Balanced Salt Solution) leaving 100 u1 of buffer after the final wash.
A stock solution of 1 mg/ml fluo-4 is made in 10% pluronic acid DMSO. To load the cells with fluo-4 , 50 u1 of 12 ug/ml fluo-4 is added to each well.
The plate is incubated at 37 degrees C in a COZ incubator for 60 min. The plate is washed four times in the Biotek washer with HBSS leaving 100 u1 of buffer.
For non-adherent cells, the cells are spun down from culture media. Cells are re-suspended to 2-Sx 106 cells/ml with HBSS ~in a 50-ml conical tube. 4 u1 of f mg/ml fluo-4 solution in 10% pluronic acid DMSO is added to each ml of cell suspension.
The tube is then placed in a 37 degrees C water bath for 30-60 min. The cells are washed twice with HBSS, resuspended to 1 x 1 O6 cells/ml, and dispensed into a microplate, 100 ul/well. The plate is centrifuged at 1000 rpm for 5 min. The plate is then washed once in Denley CellWash with 200 u1, followed by an aspiration step to 100 u1 final volume.
For a non-cell based assay, each yell contains a fluorescent molecule, such as fluo-4 . .The supernatant is added to the well, and a change in fluorescence is detected.
To measure the fluorescence of intracellular calcium; the FLIPR is set for the following parameters: (1) System gain is 300-800 mW; (2) Exposure time is 0.4 second; (3) Camera F/stop is F/2; (4) Excitation is 488 nm; (5) Emission is 530 nm;
and (6) Sample addition is 50 u1. Increased emission at 530 nm indicates an extracellular signaling event which has resulted in an increase in the intracellular Ca++ concentration.

Example 19: High-Throughput Screening Assay Identifying Tyrosine Kinase Activi The Protein Tyrosine Kinases (PTK) represent a diverse group of transmembrane and cytoplasmic kinases. Within the Receptor Protein Tyrosine Kinase RPTK) group are receptors for a range of mitogenic and metabolic growth factors including the PDGF, FGF, EGF, NGF, HGF and Insulin receptor subfamilies.
In addition there are a large family of RPTKs for which the corresponding ligand is unknown. Ligands for RPTKs include mainly secreted small proteins, but also membrane-bound and extracellular matrix proteins.
Activation of RPTK by ligands involves ligand-mediated receptor dimerization, resulting in transphosphorylation of the receptor subunits and activation of the cytoplasmic tyrosine kinases. The cytoplasmic tyrosine kinases include receptor associated tyrosine kinases of the src-family (e.g., src, yes, lck, lyn, fyn) and non-receptor linked and cytosolic protein tyrosine kinases, such as the Jak family, 1 S members of which mediate signal transduction triggered by the cytokine superfamily of receptors (e.g., the Interleukins, Interferons, GM-CSF, and Leptin).
Because of the wide range of known factors capable of stimulating tyrosine kinase activity, the identification of novel human secreted proteins capable of activating tyrosine kinase signal transduction pathways are of interest.
Therefore, the following protocol is designed to identify those novel human secreted proteins capable of activating the tyrosine kinase signal transduction pathways.
Seed target cells (e.g., primary keratinocytes) at a density of approximately.
25,000 cells per well in a 96 well Loprodyne Silent Screen Plates purchased from Nalge Nunc (Naperville, IL). The plates are sterilized with two 30 minute rinses with 100% ethanol, rinsed with water and dried overnight. Some plates are coated for 2 hr with 100 ml of cell culture grade type I collagen (50 mg/ml), gelatin (2%) or polylysine (50 mg/ml), all of which can be purchased from Sigma Chemicals (St.
Louis, MO) or 10% Matrigel purchased from Becton Dickinson (Bedford,MA), or calf serum, rinsed with PBS and stored at 4 degree C. Cell growth on these plates is assayed by seeding 5,000 cells/well in growth medium and indirect quantitation of cell number through use of alamarBlue as described by the manufacturer Alamar Biosciences, Inc. (Sacramento, CA) after 48 hr. 'Falcon plate covers #3071 from Becton Dickinson (Bedford,MA) are used to cover the Loprodyne Silent Screen Plates. Falcon Microtest III cell culture plates can also be used in some proliferation experiments.
To prepare extracts, A431 cells are seeded onto the nylon membranes of Loprodyne plates (20,000/200m1/well) and cultured overnight in complete medium.
Cells are quiesced by incubation in serum-free basal medium for 24 hr. After 5-minutes treatment with EGF (60ng/ml) or 50 u1 of the supernatant produced in Example 11, the medium was removed and 100 ml of extraction buffer ((20 mM
HEPES pH 7.5, 0.15 M NaCI, 1% Triton X-100, 0.1% SDS, 2 mM Na3V04, 2 mM
Na4P2O7 and a cocktail of protease inhibitors (# 1836170) obtained from Boeheringer Mannheim (Indianapolis, IN) is added to each well and the plate is shaken on a rotating shaker for 5 minutes at 4 degrees C. The plate is then placed in a vacuum transfer manifold and the extract filtered through the 0.45 mm membrane bottoms of each well using house vacuum. Extracts are collected in a 96-well catch/assay plate in the bottom of the vacuum manifold and immediately placed on ice. To obtain extracts clarified by centrifugation, the content of each well, after detergent solubilization for 5 minutes, is removed and centrifuged for 15 minutes at 4 degrees C at 16,000 x g.
Test the filtered extracts for levels of tyrosine kinase activity. Although many methods of detecting tyrosine kinase activity are known, one method is described here.
Generally, the tyrosine kinase activity of a supernatant is evaluated by determining its ability to phosphorylate a tyrosine residue on a specific substrate (a biotinylated peptide). Biotinylated peptides that can be used for this purpose include PSK1 (corresponding to amino acids 6-20 of the cell division kinase cdc2-p34) and PSK2 (corresponding to amino acids 1-17 of gastrin). Both peptides are substrates for a range of tyrosine kinases and are available from Boehringer Mannheim The tyrosine kinase reaction is set up by adding the following components in order. First, add 10u1 of 5uM Biotinylated Peptide, then 10u1 ATP/Mg2+ (SmM
ATP/SOmM MgCl2), then 10u1 of Sx Assay Buffer (40mM imidazole hydrochloride, pH7.3, 40 mM beta-glycerophosphate, 1mM EGTA, 100mM MgCl2, 5 mM MnCl2~

0.5 mg/ml BSA), then Sul of Sodium Vanadate(1mM), and then Sul of water. Mix the components gently and preincubate the reaction mix at 30 degrees C for 2 min.
Initial the reaction by adding 10u1 of the control enzyme or the filtered supernatant.
The tyrosine kinase assay reaction is then terminated by adding 10 u1 of 120mm EDTA and place the reactions on ice.
Tyrosine kinase activity is determined by transferring SO u1 aliquot of reaction mixture to a microtiter plate (MTP) module and incubating at 37 degrees C for min. This allows the streptavadin coated 96 well plate to associate with the biotinylated peptide. Wash the MTP module with 300u1/well of PBS four times.
Next add 75 u1 of anti-phospotyrosine antibody conjugated to horse radish peroxidase(anti-P-Tyr-POD(O.Su/ml)) to each well and incubate at 37 degrees C
for one hour. Wash the well as above.
Next add 100u1 of peroxidase substrate solution (Boehringer Mannheim) and incubate at room temperature for at least 5 mins (up to 30 min). Measure the absorbance of the sample at 405 nm by using ELISA reader. The level of bound peroxidase activity is quantitated using an ELISA reader and reflects the level of tyrosine kinase activity.
Example 20: High-Throughput Screening Assay Identifyin~ Phosphorylation Activi As a potential alternative and/or compliment to the assay of protein tyrosine kinase activity described in Example 19, an assay which detects activation (phosphorylation) of major intracellular signal transduction intermediates can also be .
used. For example, as described below one particular assay can detect tyrosine phosphorylation of the Erk-1 and Erk-2 kinases. However, phosphorylation of other molecules, such as Raf; JNK, p38 MAP, Map kinase kinase (MEK), MEK kinase, Src, Muscle specific kinase (MuSK), IRAK, Tec, and Janus, as well as any other phosphoserine, phosphotyrosine, or phosphothreonine molecule, can be detected by substituting these molecules for Erk-1 or Erk-2 in the following assay.
Specifically, assay plates are made by coating the wells of a 96-well ELISA
plate with O.lml of protein G (lug/ml) for 2 hr at room temp, (RT). The plates are then rinsed with PBS and blocked with 3% BSA/PBS for 1 hr at RT. The protein G

plates are then treated with 2 commercial monoclonal antibodies ( 1 OOng/well) against Erk-land Erk-2 (1 hr at RT) (Santa Cruz Biotechnology). (To detect other molecules, this step can easily be modified by substituting a monoclonal antibody detecting any of the above described molecules.) After 3-5 rinses with PBS, the plates are stored at 4 degrees C until use.
A431 cells are seeded at 20,000/well in a 96-well Loprodyne filterplate and cultured overnight in growth medium. The cells are then starved for 48 hr in basal medium (DMEM) and then treated with EGF (6ng/well) or 50 u1 of the supernatants obtained in Example 11 for 5-20 minutes. The cells are then solubilized and extracts filtered directly into the assay plate.
After incubation with the extract for 1 hr at RT; the wells are again rinsed.
As a positive control, a commercial preparation of MAP kinase ( l Ong/well) is used in place of A431 extract. Plates are then treated with a commercial polyclonal (rabbit) antibody (lug/ml) which specifically recognizes the phosphorylated epitope of the Erk-1 and Erk-2 kinases (1 hr at RT). This antibody is biotinylated by standard procedures. The bound polyclonal antibody is then quantitated by successive incubations with Europium-streptavidin and Europium fluorescence enhancing reagent in the Wallac DELFIA instrument (time-resolved fluorescence). An increased fluorescent signal over background indicates a phosphorylation.
Example 21: Method of Determining Alterations in a Gene Corresponding to a Polynucleotide , RNA isolated from entire families or individual patients presenting with a phenotype of interest (such as a disease) is be isolated. cDNA is then generated from these RNA samples using protocols known in the art. (See, Sambrook.) The cDNA
.
is then used as a template for PCR, employing primers surrounding regions of interest in SEQ >D NO:X. Suggested PCR conditions consist of 35 cycles at 95 degrees C
for seconds; 60-120 seconds at 52-58 degrees C; and 60-120 seconds at 70 degrees C, using buffer solutions described in Sidransky et al., Science 252:706 ( 1991 ).
30 ~ PCR products are then sequenced using primers labeled at their 5' end with T4 polynucleotide kinase, employing SequiTherm Polymerise. (Epicentre Technologies). The intron-exon borders of selected exons is also determined and _ genomic PCR products analyzed to confirm the results. PCR products harboring' suspected mutations is then cloned and sequenced to validate the results of the direct sequencing.
PCR products is cloned into T-tailed vectors as described in Holton et al., Nucleic Acids Research, 19:1156 (1991) and sequenced with T7 polymerase (United States Biochemical). Affected individuals are identified by mutations not present in unaffected individuals.
Genomic rearrangements are also observed as a method of determining alterations in a gene corresponding to a polynucleotide. Genomic clones isolated according to Example 2 are nick-translated with digoxigenindeoxy-uridine 5'-triphosphate (Boehringer Manheim), and FISH performed as described in Johnson et al., Methods Cell Biol. 35:73-99 (1991). Hybridization with the labeled probe is carried out using a vast excess of human cot-1 DNA for specific hybridization to the corresponding genoriiic locus.
Chromosomes are counterstained with 4,6-diamino-2-phenylidole and propidium iodide, producing a combination of C- and R-bands. Aligned images for precise mapping are obtained using a triple-band filter set (Chroma Technology, Brattleboro, VT) in combination with a cooled charge-coupled device camera (Photometrics, Tucson, AZ) and variable excitation wavelength filters.
(Johnson et al., Genet. Anal. Tech. Appl., 8:75 (1991).) Image collection, analysis and chromosomal fractional length measurements are performed using the ISee Graphical Program System. (Inovision Corporation, Durham, NC.) Chromosome alterations of the genomic region hybridized by the probe are identified as insertions, deletions, and translocations. These alterations are used as a diagnostic marker for an associated disease.
Example 22: Method of Detecting Abnormal Levels of a Polvnentide in a Biological Sample A polypeptide of the present invention can be detected in a biological sample, and if an increased or decreased level of the polypeptide is detected, this polypeptide is a marker for a particular phenotype. Methods of detection are numerous, and thus, it is understood that one skilled in the art can modify the following assay to fit their particular needs.
For example, antibody-sandwich ELISAs are used to detect polypeptides in a sample, preferably a biological sample. Wells of a microtiter plate are coated with specific antibodies, at a final concentration of 0.2 to l0~ug/ml. The antibodies are either monoclonal or polyclonal and are produced by the method described in Example 10. The wells are blocked so that non-specific binding of the polypeptide to the well is reduced.
The coated wells are then incubated for > 2 hours at RT with a sample containing the polypeptide. Preferably, serial dilutions of the sample should be used to validate results. The plates are then washed three times with deionized or distilled water to remove unbounded polypeptide.
Next, 50 u1 of specific antibody-alkaline phosphatase conjugate, at a concentration of 25-400 ng, is added and incubated for 2 hours at room temperature.
The plates are again washed three times with deionized or distilled water to remove unbounded conjugate.
Add 75 u1 of 4-methylumbelliferyl phosphate (MUP) or p-nitrophenyl phosphate (NPP) substrate solution to each well and incubate 1 hour at room temperature. Measure the reaction by a microtiter plate reader. Prepare a standard curve, using serial dilutions of a control sample, and plot polypeptide concentration on the X-axis (log scale) and fluorescence or absorbance of the Y-axis (linear scale).
Interpolate the concentration of the polypeptide in the sample using the standard curve.
Example 23: Formulation The invention also provides methods of treatment and/or prevention of diseases or disorders (such as, for example, any one or more of the diseases or disorders disclosed herein) by administration to a subject of an effective amount of a Therapeutic. By therapeutic is meant polynucleotides or polypeptides of the invention (including fragments and variants), agonists or antagonists thereof, and/or antibodies thereto, in combination with a pharmaceutically acceptable Garner type (e.g., a sterile Garner). _ The Therapeutic will be formulated and dosed in a fashion consistent with good medical practice, taking into account the clinical condition of the individual patient (especially the side effects of treatment with the Therapeutic alone), the site of delivery, the method of administration, the scheduling of administration, and other factors known to practitioners. The "effective amount" for purposes herein is thus determined by such considerations.
As a general proposition, the total pharmaceutically effective amount of the Therapeutic administered parenterally per dose will be in the range of about lug/kg/day to 10 mg/kg/day of patient body weight, although, as noted above, this will be subject to therapeutic discretion. More preferably, this dose is at least 0.01 mg/kg/day, and most preferably for humans between about 0.01 and 1 mg/kg/day for the hormone. If given continuously, the Therapeutic is typically administered at a dose rate of about 1 ug/kg/hour to about 50 uglkg/hour, either by 1-4 injections per day or by continuous subcutaneous infusions, for example, using a mini-pump.
An intravenous bag solution may also be employed. The length of treatment needed to observe changes and the interval following treatment for responses to occur appears to vary depending on the desired effect.
Therapeutics can be are administered orally, rectally; parenterally, intracistemally, intravaginally, intraperitoneally, topically (as by powders, ointments, gels, drops or transdermal patch), bucally, or as an oral or nasal spray.
"Pharmaceutically acceptable carrier" refers to a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any. The term .
"parenteral" as used herein refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection and infusion.
Therapeutics of the invention are also suitably administered by sustained-release systems. Suitable examples of sustained-release Therapeutics are administered orally, rectally, parenterally, intracistemally, intravaginally, intraperitoneally, topically (as by powders, ointments, gels, drops or transdermal patch), bucally, or as an oral or nasal spray. "Pharmaceutically acceptable carrier"
refers to a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. The term "parenteral" as used herein refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection and infusion.
Therapeutics of the invention are also suitably administered by sustained-release systems. Suitable examples of sustained-release Therapeutics include suitable polymeric materials (such as, for example, semi-permeable polymer matrices in the form of shaped articles, e.g., films, or mirocapsules), suitable hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, and sparingly soluble derivatives (such as, for example, a sparingly soluble salt).
Sustained-release matrices include polylactides (U.S. Pat. No. 3,773,919, EP
58,481), copolymers of L-glutamic acid and gamma-ethyl-L-glutamate (Sidman et al., Biopolymers 22:547-556 (1983)), poly (2- hydroxyethyl methacrylate) (Larger et al., J. Biomed. Mater. Res. 15:167-277 (1981), and Larger, Chem. Tech. 12:98-105 (1982)), ethylene vinyl acetate (Larger et al., Id.) or poly-D- (-)-3-hydroxybutyric acid (EP 133,988).
Sustained-release Therapeutics also include liposomally entrapped Therapeutics of the invention (see generally, Larger, Science 249:1527-1533 (1990);
Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 317 -327 and 353-365 (1989)).
Liposomes containing the Therapeutic are prepared by methods known per se: DE
3,218,121; Epstein et al., Proc. Natl. Acad. Sci. (LISA) 82:3688-3692 (1985);
Hwang et al., Proc. Natl. Acad. Sci.(USA) 77:4030-4034 (1980); EP 52,322; EP 36,676;
EP
88,046; EP 143,949; EP 142,641; Japanese Pat. Appl. 83-118008; U.S. Pat. Nos.
4,485,045 and 4,544,545; and EP 102,324. Ordinarily, the liposomes are of the small (about 200-800 Angstroms) unilamellar type in which the lipid content is greater than about 30 mol. percent cholesterol, the selected proportion being adjusted for the optimal Therapeutic.
In yet an additional embodiment, the Therapeutics of the invention are delivered by way of a pump (see Larger, supra; Sefton, CRC Crit. Ref. Biomed.
Erg.
14:201 (1987); Buchwald et al., Surgery 88:507 (.1980); Saudek et al., N.
Engl. J.
Med. 321:574 (1989)).
Other controlled release systems are discussed in the review by Larger (Science 249:1527-1533 (1990)). _ For parenteral administration, in one embodiment, the Therapeutic is formulated generally by mixing. it at the desired degree of purity, in a unit dosage injectable form (solution,'suspension, or emulsion), with a.pharmaceutically acceptable Garner, i.e., one that is non-toxic to recipients at the dosages and S concentrations employed and is compatible with other ingredients of the formulation.
For example, the formulation preferably does not include oxidizing agents and other compounds that are known to be deleterious to the Therapeutic.
Generally, the formulations are prepared by contacting the Therapeutic uniformly and intimately with liquid carriers or finely divided solid carriers or both.
Then, if necessary, the product is shaped into the desired formulation.
Preferably the carrier is a parenteral carrier, more preferably a solution that is isotonic with the blood of the recipient. Examples of such Garner vehicles include water, saline, Ringer's solution, and dextrose solution. Non-aqueous vehicles such as fixed oils and ethyl oleate are also useful herein, as well as liposomes.
The carrier suitably contains minor amounts of additives such as substances that enhance isotonicity and chemical stability. Such materials are non-toxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, succinate, acetic acid, and other organic acids or their salts;
antioxidants such as ascorbic acid; low molecular weight (less than about ten residues) polypeptides, e.g., polyarginine or tripeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids, such as glycine, glutamic acid, aspartic acid, or arginine; monosaccharides, disaccharides, and other carbohydrates including cellulose or its derivatives, glucose, manose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; counterions such as sodium;, and/or nonionic surfactants such as polysorbates, poloxamers, or PEG.
The Therapeutic is typically formulated in such vehicles at a concentration of about 0.1 mg/ml to 100 mg/ml, preferably 1-10 mg/ml, at a pH of about 3 to 8.
It will be understood that the use of certain of the foregoing excipients, carriers, or stabilizers will result in the formation of polypeptide salts.
Any pharmaceutical used for therapeutic administration can be sterile.
Sterility is readily accomplished by filtration through sterile filtration membranes.

(e.g., 0.2 micron membranes). Therapeutics generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.
Therapeutics ordinarily will be stored in unit or mufti-dose containers, for example, sealed ampoules or vials, as an aqueous solution or as a lyophilized formulation for reconstitution. As an example of a lyophilized formulation, 10-ml vials are filled with S ml of sterile-filtered 1 % (w/v) aqueous Therapeutic solution, and the resulting mixture is lyophilized. The infusion solution is prepared by reconstituting the lyophilized Therapeutic using bacteriostatic Water-for-Injection.
The invention also provides a pharmaceutical pack or kit comprising one or more containers-filled with one or more of the ingredients of the Therapeutics of the invention. Associated with such containers) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration. In addition, the Therapeutics may be employed in conjunction with other therapeutic compounds.
The Therapeutics of the inventiommay be administered alone or in combination with adjuvants. Adjuvants that may be administered with the Therapeutics of the invention include, but are not limited to, alum, alum plus deoxycholate (ImmunoAg), MTP-PE (Biocine Corp.), QS21 (Generitech, Inc.), BCG
(e.g., THERACYS~), MPL and nonviable prepartions of Corynebacterium parvum.
In a specific embodiment, Therapeutics of the invention are administered in combination with alum. In another specific embodiment, Therapeutics of the invention are administered in combination with QS-21. Further adjuvants-that may be administered with the Therapeutics of the invention include, but are not limited to, Monophosphoryl lipid immunomodulator, AdjuVax 100a, QS-21, QS-18, CRL1005, Aluminum salts, MF-59, and Virosomal adjuvant technology. Vaccines that may be administered with the Therapeutics of the invention include, but are not limited to, vaccines directed toward protection against MMR (measles, mumps, rubella), polio, varicella, tetanus/diptheria, hepatitis A, hepatitis B, haemophilus influenzae B, whooping cough, pneumonia, influenza, Lyme's Disease, rotavirus, cholera, yellow fever, Japanese encephalitis, poliomyelitis, rabies, typhoid fever, and pertussis.

Combinations may be administered either concomitantly, e.g., as an admixture, separately but simultaneously or concurrently; or sequentially. This includes presentations in which the combined agents are administered together as a therapeutic mixture, and also procedures in which the combined agents are administered separately but simultaneously, e.g., as through separate intravenous lines into the same individual. Administration "in combination" further includes the separate administration of one of the compounds or agents given first, followed by the second.
The Therapeutics of the invention may be administered alone or in combination with other therapeutic agents. Therapeutic agents that may be administered in combination with the Therapeutics of the invention, include but not limited to, chemotherapeutic agents, antibiotics, steroidal and non-steroidal anti=
inflammatories, conventional immunotherapeutic agents, and/or therapeutic treatments described below. Combinations may be administered either concomitantly, e.g., as an admixture, separately but simultaneously or concurrently;
of sequentially. This includes presentations in which the combined agents are administered together as a therapeutic mixture, and'also procedures in which the combined agents are administered separately but simultaneously, e.g., as through separate intravenous lines into the same individual. Administration "in combination"
further includes the separate administration of one of the compounds or agents given first, followed by the second.
In certain embodiments, Therapeutics of the invention are administered in combination with antiretroviral agents, nucleoside/nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), and/or protease inhibitors (PIs). NRTIs that may be administered in combination with the Therapeutics of the invention, include, but are not limited to, RETROVIRT""
(zidovudine/AZT), VIDEXT"" (didanosine/ddI), HIVIDT"~ (zalcitabine/ddC), ZERITT""
(stavudine/d4T), EPNIRT"" (lamivudine/3TC), and COMBNIRT""
(zidovudine/lamivudine). NNRTIs that may be administered in combination with the Therapeutics of the invention, include, but are not limited to, VIRAMUNET"~
(nevirapine), RESCRIPTORT"" (delavirdine), and SUSTNAT"" (efavirenz). Protease inhibitors that may be administered in combination with the Therapeutics of the invention, include, but are not limited to, CRIXIVANT"" (indinavir), NORVIRT""
(ritonavir), INVIRASET"" (saquinavir), and VIRACEPTT"" (nelfinavir). In a specific embodiment, antiretroviral agents, nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, and/or protease inhibitors may be used in any combination with Therapeutics of the invention to treat AIDS and/or to prevent or treat HIV infection. , Additional NRTIs include LODENOSINET"" (F-ddA; an acid-stable adenosine NRTI; Triangle/Abbott; COVIRACILT"" (emtricitabine/FTC; structurally related to lamivudine (3TC) but with 3- to 10-fold greater activity in vitro;
Triangle/Abbott);
dOTC (BCH-10652, also structurally related to lamivudine but retains activity against a substantial proportion of lamivudine-resistant isolates; Biochem Pharma);
Adefovir (refused approval for anti-HIV therapy by FDA; Gilead Sciences);
PREVEON~ (Adefovir Dipivoxil, the active prodrug of adefovir; its active form is PMEA-pp); TENOFOVIRT"" (bis-POC PMPA, a PMPA prodrug; Gilead);
DAPD/DXG (active metabolite of DAPD; Triangle/Abbott); D-D4FC (related to 3TC, with activity against AZT/3TC-resistant virus); GW420867X (Glaxo Wellcome); ZIAGENT"' (abacavir/159U89; Glaxo Wellcome Inc.); CS-87 (3'azido-2',3'-dideoxyuridine; WO 99/66936); and S-acyl-2-thioethyl (SATE)-bearing prodrug forms of (3-L-FD4C and ~3-L-FddC (WO 98/17281).
Additional NNRTIs include COACTINONT"" (Emivirine/MKC-442, potent NIVRTI of the HEPT class; Triangle/Abbott); CAPRAVIRINET"" (AG-1549/S-1153, a next generation NNRTI with activity against viruses,containing the K103N
mutation; Agouron); PNU-142721 (has 20- to 50-fold greater activity than its predecessor delavirdine and is active against K103N mutants; Pharmacia &
Upjohn);
DPC-961 and DPC-963 (second-generation derivatives of efavirenz, designed to be active against viruses with the K103N mutation; DuPont); GW-420867X' (has 25-fold greater activity than HBY097 and is active against K103N mutants; Glaxo Wellcome); CALANOLIDE A (naturally occurring agent from the latex tree; active against viruses containing either or both the Y181C and K103N mutations); and Propolis (WO 99/49830).

Additional protease inhibitors include LOPINAVIRT"" (ABT378/r; Abbott Laboratories); BMS-232632 (an azapeptide; Bristol-Myres Squibb); TIPRANAVIRT""
(PNU-140690, a non-peptic dihydropyrone; Pharmacia & Upjohn); PD-178390 (a nonpeptidic dihydropyrone; Parke-Davis); BMS 232632 (an azapeptide; Bristol-Myers Squibb); L-756,423 (an indinavir analog; Merck); DMP-450 (a cyclic urea compound; Avid & DuPont); AG-1776 (a peptidomimetic with in vitro activity against protease inhibitor-resistant viruses; Agouron); VX-175/GW-433908 (phosphate prodrug of amprenavir; Vertex & Glaxo Welcome); CGP61755 (Ciba);
and AGENERASET"" (amprenavir; Glaxo Wellcome Inc.).
Additional antiretroviral agents include fusion inhibitors/gp41 binders.
Fusion inhibitors/gp41 binders include T-20 (a peptide from residues 643-678 of the HIV gp41 transmembrane protein ectodomain which binds to gp41 in its resting state and prevents transformation to the fusogenic state; Trimeris) and T-1249 (a second-generation fusion inhibitor; Trimeris).
Additional antiretroviral agents include fusion inhibitors/chemokine receptor antagonists. Fusion inhibitors/chemokine receptor antagonists include CXCR4 antagonists such as AMD 3100 (a bicyclam), SDF-1 and its analogs, and ALX40-4C
(a cationic peptide), T22 (an 18 amino acid peptide; Trimeris) and the T22 analogs T134 and T140; CCRS antagonists such as RANTES (9-68), AOP-RANTES, NNY-RANTES, and TAK-779; and CCRS/CXCR4 antagonists such as NSC 651016 (a distamycin analog). Also included are CCR2B, CCR3; and CCR6 antagonists.
Chemokine recpetor agonists such as RANTES, SDF-1, MIP-la, MIP-1(3, etc., may also inhibit fusion.
Additional antiretroviral agents include integrase inhibitors. Integrase , inhibitors include dicaffeoylquinic (DFQA) acids; L-chicoric acid (a dicaffeoyltartaric (DCTA) acid); quinalizarin (QLC) and related anthraquinones;
ZINTEVIRT"" (AR 177, an oligonucleotide that probably acts at cell surface rather than being a true integrase inhibitor; Arondex); and naphthols such as those disclosed in WO 98/50347.
Additional antiretroviral agents include hydroxyurea-like compunds such as BCX-34 (a purine nucleoside phosphorylase inhibitor; Biocryst); ribonucleotide reductase inhibitors such as DIDOXT"" (Molecules for Health); inosine monophosphate dehydrogenase (IMPDH) inhibitors sucha as VX-497 (Vertex); and mycopholic acids such as CellCept (mycophenolate mofetil; Roche).
Additional antiretroviral agents include inhibitors of viral integrase, inhibitors of viral genome nuclear translocation such as arylene bis(methylketone) compounds;
inhibitors of HN entry such as AOP-RANTES, NNY-RANTES, RANTES-IgG
fusion protein, soluble complexes of RANTES and glycosaminoglycans (GAG), and AMD-3100; nucleocapsid zinc finger inhibitors such as dithiane compounds;
targets of HIV Tat and Rev; and pharmacoenhancers such as ABT-378.
Other antiretroviral therapies and adjunct therapies include cytokines and lymphokines such as MIP-la, MIP-1(3; SDF-la, IL-2,°PROLEUKINT""
(aldesleukin/L2-7001; Chiron), IL-4, IL-10, IL-12, and IL-13; interferons such as IFN-a2a; antagonists of TNFs, NFxB, GM-CSF, M-CSF, and IL-10; agents that modulate immune activation such as cyclosporin and prednisone; vaccines such as RemuneT"" (HIV Immunogen), APL 400-003 (Apollon), recombinant gp120 and fragments, bivalent (B/E) recombinant envelope glycoprotein; rgp120CM235, MN
rgp120, SF-2 rgp120, gp120/soluble CD4 complex, Delta JR-FL protein, branched synthetic peptide derived from discontinuous gp120 C3/C4 domain, fusion-competent immunogens, and Gag, Pol, Nef, and Tat vaccines; gene-based therapies such as genetic suppressor elements (GSEs;, WO 98/54366), and intrakines (genetically modified CC chemokines targetted to the ER to block surface expression of newly synthesized CCRS (Yang et al., PNAS 94:11567-72 (1997); Chen et al., Nat. Med. 3:1110-16 (1997)); antibodies such as the anti-CXCR4 antibody 1265, the anti-CCRS antibodies 2D7, 5C7, PAB, PA9, PA10, PA11, PA12, and PA14, the anti-CD4 antibodies Q4120 and RPA-T4, the anti-CCR3 antibody 7B 11, the anti-gp 120 antibodies 17b, 48d, 447-52D, 257-D, 268-D and 50.1, anti-Tat antibodies, anti-TNF-a antibodies, and monoclonal antibody 33A; aryl hydrocarbon (AH) receptor agonists and antagonists such as TCDD, 3,3',4,4',5-pentachlorobiphenyl, 3,3',4,4'-tetrachlorobiphenyl, and a-naphthoflavone (WO 98/30213); and antioxidants such as y-L-glutamyl-L-cysteine ethyl ester (y-GCE; WO 99/56764).

In a further embodiment, the Therapeutics of the invention are administered in combination with an antiviral agent. Antiviral agents that may be administered with the Therapeutics of the invention include, but are not limited to, acyclovir, ribavirin, amantadine, and remantidine.
In other embodiments, Therapeutics of the invention may be administered in combination with anti-opportunistic infection agents. Anti-opportunistic agents that may be administered in combination with the Therapeutics of the invention, include, but are not limited to, TRIMETHOPRIM-SULFAMETHOXAZOLET"", DAPSONET"", PENTAMIDINET"', ATOVAQUONET"", ISONIAZIDT"', RIFAMPINT"", PYRAZINAMIDET"~, ETHAMBUTOLT"", RIFABUTINT"", .
CLARITHROMYCINT"", AZITHROMYCINT"", GANCICLOVIRT"", FOSCARNETT"~, CIDOFOVIRT"", FLUCONAZOLET"", ITRACONAZOLET"", KETOCONAZOLET"", ACYCLOVIRT"", FAMCICOLVIRT"", PYRIMETHAMINET"~, LEUCOVORINT"", NEUPOGENT"" (filgrastim/G-CSF), and LEUKINET""
(sargramostim/GM-CSF). In a specific embodiment, Therapeutics of the invention are used in any combination with TRIMETHOPRIM-SULFAMETHOXAZOLET"", DAPSbNET"", PENTAMIDINET"", and/or ATOVAQUONET"" to prophylactically treat or prevent an opportunistic Pneumocystis carinii pneumonia infection. In another specific embodiment, Therapeutics of the invention are used in any combination with ISONIAZIDT"", R1FAMPINT"", PYRAZINAMIDET"", and/or ETHAMBUTOLT"" to prophylactically treat or prevent an opportunistic Mycobacterium avium complex infection. In another specific embodiment, Therapeutics of the invention are used in any combination with RIFABUTINT"", CLARITHROMYCINT"", and/or AZITHROMYC1NT"" to prophylactically treat or prevent an opportunistic Mycobacterium tuberculosis infection. In another specific embodiment, Therapeutics of the invention are used in any combination with GANCICLOVIRT"", FOSCARNETT"", and/or CIDOFOVIRT"" to prophylactically treat or prevent an opportunistic cytomegalovirus infection. In another specific embodiment, Therapeutics of the invention are used in any combination with FLUCONAZOLET"", ITRACONAZOLET"", and/or KETOCONAZOLET"' to prophylactically treat or prevent an opportunistic fungal infection. In another specific embodiment, Therapeutics of the invention are used in any combination with ACYCLOVIRT"~ and/or FAMCICOLVIRTM to prophylactically treat or prevent an opportunistic herpes simplex virus type, I and/or type II infection. In another specific embodiment, Therapeutics of the invention are used in any combination with PYRIMETHAMINET"" and/or LEUCOVORINT"" to prophylactically treat or prevent an opportunistic Toxoplasma gondii infection. In another specific embodiment, Therapeutics of the invention are used in any combination with LEUCOVORINT""
and/or NEUPOGENT"" to prophylactically treat or prevent an opportunistic bacterial infection.
In a further embodiment, the Therapeutics of the invention are administered in combination with an antibiotic agent. Antibiotic agents that may be administered with the Therapeutics of the invention include, but are not limited to, amoxicillin, beta-lactamases, aminoglycosides, beta-lactam (glycopeptide), beta-lactamases, Clindamycin, chloramphenicol, cephalosporins, ciprofloxacin, erythromycin, fluoroquinolones, macrolides, metronidazole, penicillins, quinolones, rapamycin, rifampin, streptomycin, sulfonamide, tetracyclines, trimethoprim, trimethoprim-sulfamethoxazole, and vancomycin. .
In other embodiments, Therapeutics of the invention are administered in combination with immunosuppressive agents. Immunosuppressive agents that may be administered'in combination with the Therapeutics of the invention include,. but are not limited to, steroids, cyclosporine, cyclosporine analogs, cyclophosphamide methylprednisone, prednisone, azathioprine, FK-506, 15-deoxyspergualin, and other immunosuppressive, agents that act by suppressing the function of responding T
cells.
Other immunosuppressive agents that may be administered in combination with the Therapeutics of the invention include, but are not limited to, prednisolone, methotrexate, thalidomide, methoxsalen, rapamycin, leflunomide, mizoribine (BREDININTM), brequinar, deoxyspergualin, and azaspirane (SKF 105685), ORTHOCLONE OKT~ 3 (muromonab-CD3), SANDIMMUNET"", NEORALT"", SANGDYAT"" (cyclosporine), PROGRAF~ (FK506, tacrolimus), CELLCEPT~
(mycophenolate motefil, of which the active metabolite is mycophenolic acid), IMURANTM (azathioprine), glucocorticosteroids, adrenocortical steroids such as DELTASONETM (prednisone) and HYDELTRASOLTM (prednisolone), FOLEXTM
and MEXATETM (methotrxate), OXSORALEN-ULTR.ATM (methoxsalen) and ~TM (sirolimus). ~ In a specific embodiment, immunosuppressants may be used to prevent rejection of organ or bone marrow transplantation.
~In an additional embodiment, Therapeutics of the invention are administered alone or in combination with one or more intravenous immune globulin preparations.
Intravenous immune globulin preparations that may be administered with the Therapeutics of the invention include, but not limited to, GAMMARTM, IVEEGAMT"", SANDOGLOBULINT"", GAMMAGARD S/DT"~, ATGAMTM
(antithymocyte glubulin), and GAMIMUNET"". In a specific embodiment, Therapeutics of the invention are administered in combination with intravenous immune globulin preparations in transplantation therapy (e.g., bone marrow transplant). , In certain embodiments, the Therapeutics of the invention are administered alone or in combination with an anti-inflammatory agent. Anti-inflammatory agents that may be administered with the Therapeutics of the invention include, but are not limited to, corticosteroids (e.g. betamethasone, budesonide, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, and triamcinolone), nonsteroidal anti-inflammatory drugs (e.g., diclofenac, diflunisal, etodolac, fenoprofen, floctafenine, flurbiprofen, ibuprofen, indomethacin, ketoprofen, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, phenylbutazone, piroxicam, sulindac, tenoxicam, tiaprofenic acid, and tolmetin.), as well as antihistamines, aminoarylcarboxylic acid derivatives, arylacetic acid derivatives, arylbutyric acid derivatives, arylcarboxylic acids, arylpropionic acid derivatives, pyrazoles, pyrazolones, salicylic acid derivatives, thiazinecarboxamides, e-acetamidocaproic acid, S-adenosylmethionine, 3-amino-4-hydroxybutyric acid, amixetrine, bendazac, benzydamine, bucolome, difenpiramide, ditazol, emorfazone, guaiazulene, nabumetone, nimesulide, orgotein, oxaceprol, paranyline, perisoxal, pifoxime, proquazone, proxazole, and tenidap.
, In an additional embodiment, the compositions of the invention are administered alone or in combination with an anti-angiogenic agent. Anti-angiogenic agents that may be administered with the compositions of the invention include, but are not limited to, Angiostatin (Entremed, Rockville, MD), Troponin-1 (Boston Life Sciences, Boston, MA), anti-Invasive Factor, retinoic acid and derivatives thereof, paclitaxel (Taxol), Suramin, Tissue Inhibitor of Metalloproteinase-1, Tissue Inhibitor of Metalloproteinase-2, VEGI, Plasminogen Activator Inhibitor-1, Plasminogen Activator Inhibitor-2, and various forms of the lighter "d group" transition metals.
Lighter "d group" transition metals include, for example, vanadium, molybdenum, tungsten, titanium, niobium, and tantalum species. Such transition metal species may form transition metal complexes. Suitable complexes of the above-mentioned transition metal species include oxo transition metal complexes.
Representative examples of vanadium complexes include oxo vanadium complexes such as vanadate and vanadyl complexes. Suitable vanadate complexes include metavanadate and orthovanadate complexes such as, for example, ammonium metavanadate, sodium metavanadate, and sodium orthovanadate. Suitable vanadyl complexes include, for example, vanadyl acetylacetonate and vanadyl sulfate including vanadyl sulfate hydrates such as vanadyl sulfate mono- and trihydrates.
Representative examples of tungsten and molybdenum complexes also include oxo complexes. Suitable oxo tungsten complexes include tungstate and tungsten oxide complexes. Suitable tungstate complexes include ammonium tungstate, calcium tungstate, sodium tungstate dihydrate, and tungstic acid. Suitable tungsten oxides include tungsten (IV) oxide and tungsten (VI) oxide. Suitable oxo molybdenum complexes include molybdate, molybdenum oxide, and molybdenyl complexes. Suitable molybdate complexes include ammonium molybdate and its _ hydrates, sodium molybdate and its hydrates, and potassium molybdate and its hydrates. Suitable molybdenum oxides include molybdenum (VI) oxide, molybdenum (VI) oxide, and molybdic acid. Suitable molybdenyl complexes include, for example, molybdenyl acetylacetonate. Other suitable tungsten and molybdenum complexes include hydroxo derivatives derived from, for example, glycerol, tartaric acid, and sugars.
A wide variety.of other anti-angiogenic factors may also be utilized within the ' context of the present invention. Representative examples include, but are not limited to, platelet factor 4; protamine sulphate; sulphated chitin derivatives (prepared from queen crab shells), (Murata et al., Cancer Res. 51:22-26, ( 1991 )); Sulphated Polysaccharide Peptidoglycan Complex (SP- PG) (the function of this compound may be enhanced by the presence of steroids such as estrogen, and tamoxifen citrate);
Staurosporine; modulators of matrix metabolism, including for example, proline analogs, cishydroxyproline, d,L-3,4-dehydroproline, Thiaproline, alpha,alpha-dipyridyl, aminopropionitrile fumarate; 4-propyl-5-(4-pyridinyl)-2(3H)-oxazolone;
Methotrexate; Mitoxantrone; Heparin; Interferons; 2 Macroglobulin-serum; ChIMP-(Pavloff et al., J. Bio. Chem. 267:17321-17326, (1992)); Chymostatin (Tomkinson et al., Biochem J. 286:475-480, (1992)); Cyclodextrin Tetradecasulfate;
Eponemycin;
Camptothecin; Fumagillin (Ingber et al., Nature 348:555-557, (1990)); Gold Sodium Thiomalate ("GST"; Matsubara and Ziff, J. Clin. Invest. 79:1440-1f46, (1987));
anticollagenase-serum; alpha2-antiplasmin (Holmes et al., J. Biol. Chem.
262(4):1659-1664, (1987)); Bisantrene (National Cancer Institute); Lobenzarit disodium (N-(2)-carboxyphenyl-4- chloroanthronilic acid disodium or "CCA";
(Takeuchi et al., Agents Actions 36:312-316, (1992)); and metalloproteinase inhibitors such as BB94.
Additional anti-angiogenic factors that may also be utilized within the context of the present invention include Thalidomide, (Celgene, Warren, NJ);
Angiostatic steroid; AGM-1470 (H. Brem and J. Folkman JPediatr. Surg. 28:445-51 (1993));
an integrin alpha v beta 3 antagonist (C. Storgard et al., J Clin. Invest. 103:47-(1999)); carboxynaminolmidazole; Carboxyamidotriazole (CAI) (National Cancer Institute, Bethesda, MD); Conbretastatin A-4 (CA4P) (OXiGENE, Boston, MA); ' Squalamine (Magainin Pharmaceuticals, Plymouth Meeting, PA); TNP-470, (Tap Pharmaceuticals, Deerfield, IL); ZD-0101 AstraZeneca (London, LJK); APRA
(CT2584); Benefin, Byrostatin-1 (5.C339555); CGP-41251 (PKC 412); CM101;
Dexrazoxane (ICRF187); DMXAA; Endostatin; Flavopridiol; Genestein; GTE;
ImmTher; Iressa (ZD1839); Octreotide (Somatostatin); Panretin; Penacillamine;
Photopoint; PI-88; Prinomastat (AG-3340) Purlytin; Suradista (FCE26644);
Tamoxifen (Nolvadex); Tazarotene; Tetrathiomolybdate; Xeloda (Capecitabine);
and 5-Fluorouracil.
Anti-angiogenic agents that may be administed in combination with the compounds of the invention may work through a variety of mechanisms including, but not limited to, inhibiting proteolysis of the extracellular matrix, blocking the function of endothelial cell-extracellular matrix adhesion molecules, by antagonizing the function of angiogenesis inducers such as growth factors, and inhibiting integrin receptors expressed on proliferating endothelial cells. Examples of anti-angiogenic inhibitors that interfere with extracellular matrix proteolysis and which may be administered in combination with the compositons of the invention include, but are not united to, AG-3340 (Agouron, La Jolla, CA), BAY-12-9566 (Bayer, West Haven, CT), BMS-275291 (Bristol Myers Squibb, Princeton, NJ), CGS-27032A (Novartis, East Hanover, NJ), Marimastat (British Biotech, Oxford, UK), and Metastat (Aeterna, St-Foy, Quebec). Examples of anti-angiogenic inhibitors that act by blocking the function of endothelial cell-extracellular matrix adhesion molecules and which may be administered in combination with the compositons of the invention include, but are not lmited to, EMD-121974, (Merck KcgaA Darmstadt, Germany) and Vitaxin (Ixsys, La Jolla, CA/Medimmune, Gaithersburg, MD). Examples of anti-angiogenic agents that act by directly antagonizing or inhibiting angiogenesis inducers and which may be administered in combination with the compositons of the invention include, but are not lmited to, Angiozyme (Ribozyme, Boulder, CO), Anti-VEGF antibody (Genentech, S. San Francisco, CA), PTK-787/ZK-225846 (Novartis, Basel, Switzerland), SU-101 (Sugeil, S. San Francisco, CA), SU-5416 (Sugen/ Pharmacia Upjohn, Bridgewater, NJ), and SU-6668 (Sugen). Other anti-angiogenic agents act to indirectly inhibit angiogenesis. Examples of indirect inhibitors of angiogenesis which may be administered in combination with the compositons of the invention include, but are not limited to, IM-862 (Cytran, Kirkland, WA), Interferon-alpha, IL-12 (Roche, Nutley, NJ), and Pentosan polysulfate (Georgetown University, Washington;
DC). ' In particular embodiments, the use of compositions of the invention in combination with anti-angiogenic agents is contemplated for the treatment, prevention, and/or amelioration of an autoimmune disease, such as for example, an autoimmune disease described herein.
In a particular embodiment, the use of compositions of the invention in combination with anti-angiogenic agents is, contemplated for the treatment, prevention, and/or amelioration of arthritis. In a more particular embodiment, the use of compositions of the invention in combination with anti-angiogenic agents is contemplated for the treatment, prevention, and/or amelioration of rheumatoid arthritis.
In another embodiment, the polynucleotides encoding a polypeptide of the present invention are administered in combination with an angiogenic protein, or S polynucleotides encoding an angiogenic protein. Examples of angiogenic proteins that may be administered with the compositions of the invention include, but are not limited to, acidic and basic fibroblast growth factors, VEGF-1, VEGF-2, VEGF-3, epidermal growth factor alpha and beta, platelet-derived endothelial cell growth factor, platelet-derived growth factor, tumor necrosis factor alpha, hepatocyte growth factor, insulin-like growth factor, colony stimulating factor, macrophage colony stimulating factor, granulocyte/macrophage colony stimulating factor, and nitric oxide synthase.
In additional embodiments, compositions of the invention are administered in combination with a chemotherapeutic agent. Chemotherapeutic agents that may be administered with the Therapeutics of the invention include, but are not limited to alkylating agents such as nitrogen mustards (for example, Mechlorethamine, cyclophosphamide, Cyclophosphamide Ifosfamide, Melphalan (L-sarcolysin), and Chlorambucil), ethylenimines and methylmelamines (for example, Hexamethylmelamirie and Thiotepa), alkyl sulfonates (for example, Busulfan), nitrosoureas (for example, Carmustine (BCNU), Lomustine (CCNL~, Semustine (methyl-CCNU), and Streptozocin (streptozotocin)), triazenes (for example, Dacarbazine (DTIC; dimethyltriazenoimidazolecarboxamide)), folic acid analogs (for example, Methotrexate (amethopterin)), pyrimidine analogs (for example, Fluorouacil (5-fluorouracil; 5-FU), Floxuridine (fluorodeoxyuridine; FudR), and Cytarabine (cytosine arabinoside)), purine analogs and related inhibitors (for example, Mercaptopurine (6-mercaptopurine-; 6-MP), Thioguanine (6-thioguanine;
TG), and Pentostatin (2'-deoxycoformycin)), vinca alkaloids (for example, Vinblastine (VLB, vinblastine sulfate)) and Vincristine (vincristine sulfate)), epipodophyllotoxins (for example, Etoposide and Teniposide), antibiotics (for example, Dactinomycin (actinomycin D), Daunorubicin (daunomycin; rubidomycin), Doxorubicin, Bleomycin, Plicamycin (mithramycin), and Mitomycin (mitomycin C), enzymes (for example, L-Asparaginase), biological response modifiers (for example, Interferon-alpha and interferon-alpha-2b), platinum coordination compounds (for example, Cisplatin (cis-DDP) and Carboplatin), anthracenedione (Mitoxantrone), substituted ureas (for example, Hydroxyurea), methylhydrazine derivatives (for example, Procarbazine (N-methylhydrazine; MIH), adrenocorticosteroids (for example, Prednisone), progestins (for example, Hydroxyprogesterone caproate, Medroxyprogesterone, Medroxyprogesterone acetate, and Megestrol acetate), estrogens (for example, Diethylstilbestrol (DES), Diethylstilbestrol diphosphate, Estradiol, and Ethinyl estradiol), antiestrogens (for example, Tamoxifen), androgens (Testosterone proprionate, and Fluoxymesterone), antiandrogens (for example, Flutamide), gonadotropin-releasing horomone analogs (for example, Leuprolide), other hormones and hormone analogs (for example, methyltestosterone, estramustine, estramustine phosphate sodium, chlorotrianisene, and testolactone), and others (for example, dicarbazine, glutamic acid, and mitotane).
In one embodiment, the compositions of the invention are administered in combination with one or more of the'following drugs: infliximab (also known as RemicadeTM Centocor, Inc.), Trocade (Roche, RO-32-3555), Leflunomide (also known as AravaTM from Hoechst Marion Roussel), KineretTM (an IL-1 Receptor antagonist also known as Anakinra from Amgen, Inc.) In a specific embodiment, compositions of the invention are administered in combination with CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) or combination of one or more of the components of CHOP. In one embodiment, the compositions of the invention are administered in combination with anti-CD20 antibodies, human monoclonal anti-CD20 antibodies. In another embodiment, the compositions of the invention are administered in combination with anti-CD20 antibodies and CHOP, or anti-CD20 antibodies and any combination of one or more of the components of CHOP, particularly cyclophosphamide and/or prednisone. In a specific embodiment, compositions of the invention are administered in combination with Rituximab. In a further embodiment, compositions of the invention are administered with Rituximab and CHOP, or Rituximab and any combination of one or more of the components of CHOP, particularly cyclophosphamide and/or prednisone. In a specific embodiment, compositions of the invention are administered in combination with tositumomab. In a further embodiment, compositions of the invention are administered with tositumomab and CHOP, or tositumomab and any combination of one or more of the components of CHOP, particularly cyclophosphamide and/or prednisone. The anti-CD20 antibodies may optionally be associated with radioisotopes, toxins or cytotoxic prodrugs.
In another specific embodiment, the compositions of the invention are administered in combination ZevalinT"". In a further embodiment, compositions of the invention are administered with ZevalinT"" and CHOP, or ZevalinT"" and any combination of one or more of the components of CHOP, particularly cyclophosphamide and/or prednisone. ZevalinT"" may be associated with one or more radisotopes. Particularly preferred isotopes are 9°Y and 1' ~In.
In an additional embodiment, the Therapeutics of the invention are administered in combination with cytokines. Cytokines that may be administered with the Therapeutics of the invention include, but are not limited to, IL2, IL3, IL4, ILS, IL6, IL7, IL 10, IL 12, IL 13, IL 1 S, anti-CD40, CD40L, IFN-gamma and TNF-alpha. In another embodiment, Therapeutics of the invention may be administered with any interleukin, including, but not limited to, IL-lalpha, IL-lbeta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19, IL-20, and IL-21.
In one embodiment, the Therapeutics of the invention are administered in combination with members of the TNF family. TNF, TNF-related or TNF-like molecules that may be administered with the Therapeutics of the invention include, but are not limited to, soluble forms of TNF-alpha, lymphotoxin-alpha (LT-alpha, also known as TNF-beta), LT-beta (found in complex heterotrimer LT-alpha2-beta), OPGL, Fast, CD27L, CD30L, CD40L, 4-1BBL, DcR3, OX40L, TNF-gamma (International Publication No. WO 96/14328), AIM-I (International Publication No.
WO 97/33899), endokine-alpha (International Publication No. WO 98/07880), OPG, and neutrokine-alpha (International Publication No. WO 98/18921, OX40, and nerve growth factor (NGF), and soluble forms of Fas, CD30, CD27, CD40 and 4-IBB, TR2 (International Publication No. WO 96/34095), DR3 (International Publication No.
WO 97/33904), DR4 (International Publication No. WO 98/32856), TRS
(International Publication No. WO 98/30693), TRANK, TR9 (International Publication No. WO 98/56892),TR10 (International Publication No. WO 98/54202), -312C2 (International Publication No. WO 98/06842), and TR12, and soluble forms CD154, CD70, and CD153.
In an additional embodiment, the Therapeutics of the invention are administered in combination with angiogenic proteins. Angiogenic proteins that may S be administered with the Therapeutics of the invention include, but are not limited to, Glioma Derived Growth Factor (GDGF), as disclosed in European Patent Number EP-399816; Platelet Derived Growth Factor-A (PDGF-A), as disclosed in European Patent Number EP-682110; Platelet Derived Growth Factor-B (PDGF-B), as disclosed in European Patent Number EP-282317; Placental Growth Factor (P1GF), as disclosed in International Publication Number WO 92/06194; Placental Growth Factor-2 (P1GF-2), as disclosed in Hauser et al., Growth Factors, 4:259-268 (1993);
Vascular Endothelial Growth Factor (VEGF), as disclosed in International Publication Number WO 90/13649; Vascular Endothelial Growth Factor-A (VEGF-A), as disclosed in European Patent Number EP-506477; Vascular Endothelial Growth Factor-2 (VEGF-2), as disclosed in International Publication Number WO
96/39515;
Vascular Endothelial Growth Factor B (VEGF-3); Vascular Endothelial Growth Factor B-186 (VEGF-B186), as disclosed in International Publication Number WO
96/26736; Vascular Endothelial Growth Factor-D (VEGF-D), as disclosed in International Publication Number WO 98/02543; Vascular Endothelial Growth Factor-D (VEGF-D), as disclosed in International Publication Number WO
98/07832;
and Vascular Endothelial Growth Factor-E (VEGF-E), as disclosed in German Patent Number DE19639601. The above mentioned references are herein incorporated by reference in their entireties.
In an additional embodiment, the Therapeutics of the invention are administered in combination with Fibroblast Growth Factors. Fibroblast Growth Factors that may be administered with the Therapeutics of the invention include, but are not limited to, FGF-1, FGF-2, FGF-3, FGF-4, FGF-5, FGF-6, FGF-7, FGF-8, FGF-9, FGF-10, FGF-11, FGF-12, FGF-13, FGF-14, and FGF-15.
In an additional embodiment, the Therapeutics of the invention are administered in combination with hematopoietic growth factors. Hematopoietic growth factors that may be administered with the Therapeutics of the invention include, but are not limited to, granulocyte macrophage colony stimulating factor (GM-CSF) (sargramostim, LEZJKINETM, PROKINETM), granulocyte colony stimulating factor (G-CSF) (filgrastim, NEUPOGENT"~), macrophage colony stimulating factor (M-CSF, CSF-1) erythropoietin (epoetin alfa, EPOGENTM, PROCRITTM), stem cell factor (SCF, c-kit ligand, steel factor), megakaryocyte colony stimulating factor, PIXY321 (a GMCSF/IL-3 fusion protein), interleukins, especially any one or more of IL-1 through IL-12, interferon-gamma, or thrombopoietin.
In certain embodiments, Therapeutics of the present invention are administered in combination with adrenergic Mockers, such as, for example, acebutolol, atenolol, betaxolol, bisoprolol, carteolol, labetalol, metoprolol, nadolol, oxprenolol, penbutolol, pindolol, propranolol, sotalol, and timolol.
In another embodiment, the Therapeutics of the invention are administered in combination with an antiarrhythmic drug (e.g., adenosine, amidoarone, bretylium, digitalis, digoxin, digitoxin, diliazem, disopyramide, esmolol, flecainide, lidocaine, mexiletine, moricizine, phenytoin, procainamide, N-acetyl procainamide, propafenone, propranolol, quinidine, sotalol, tocainide, and verapamil).
In another embodiment, the Therapeutics of the invention are administered in combination with diuretic agents, such as carbonic anhydrase-inhibiting agents (e.g., acetazolamide, dichlorphenamide, and methazolamide), osmotic diuretics (e.g., glycerin, isosorbide, mannitol, and urea), diuretics that inhibit Na+-K+-2Ch symport (e.g., furosemide, bumetanide, azosemide, piretanide, tripamide, ethacrynic acid, muzolimine, and torsemide), thiazide and thiazide-like diuretics (e.g., bendroflumethiazide; benzthiazide, chlorothiazide, hydrochlorothiazide, hydroflumethiazide, methyclothiazide, polythiazide, trichormethiazide, chlorthalidone, indapamide; metolazone, and quinethazone), potassium sparing diuretics (e.g., amiloride and triamterene), and mineralcorticoid receptor antagonists (e.g., spironolactone, canrenone, and potassium canrenoate).
In one embodiment, the Therapeutics of the invention are administered in combination with treatments for endocrine and/or hormone imbalance disorders.
Treatments for endocrine and/or hormone imbalance disoi-ders include, but are not limited to, 1271, radioactive isotopes of iodine such as i3~I and ~23I;
recombinant growth hormone, such as HLTMATROPET"" (recombinant somatropin); growth hormone analogs such as PROTROPINT"" (somatrem); dopamine agonists such as PARLODELT"" (bromocriptine); somatostatin analogs such as SANDOSTATINT""
(octreotide); gonadotropin preparations such as PREGNYLT"", A.P.L.T~~ and PROFASIT"" (chorionic gonadotropin (CG)), PERGONALT"" (menotropins), and METROD1NT"" (urofollitropin (uFSH)); synthetic human gonadotropin releasing hormone preparations such as FACTRELT"" and LUTREPULSET"" (gonadorelin hydrochloride); synthetic gonadotropin agonists such as LUPRONT"" (leuprolide acetate), SUPPRELINT"" (histrelin acetate), SYNARELT"" (nafarelin acetate), and ZOLADEXT~" (goserelin acetate); synthetic preparations of thyrotropin-releasing hormone such as RELEFACT TRHT"" and THYPINONET"" (protirelin); recombinant human TSH such as THYROGENT""; synthetic preparations of the sodium salts of the natural isomers of thyroid hormones such as L-T4T"", SYNTHROIDT"" and LEVOTHROIDT"" (levothyroxine sodium), L-T3T"', CYTOMELT"" and TRIOSTATT""
(liothyroine sodium), and THYROLART"~ (liotrix); antithyroid compounds such as n-propylthiouracil (propylthiouracil), 1-methyl-2-mercaptoimidazole and TAPAZOLET"" (methimazole), NEO-MERCAZOLET"" (carbimazole); beta-adrenergic receptor antagonists such as propranolol and esmolol; Ca2+ channel blockers;
dexamethasone and iodinated radiological contrast agents such as TELEPAQUET""
(iopanoic acid) and ORAGRAFINT"" (sodium ipodate).
Additional treatments for endocrine and/or hormone imbalance disorders include, but are not limited to, estrogens or congugated estrogens such as ESTRACET"" (estradiol), ESTINYLT"" (ethinyl estradiol), PREMARINT"", ESTRATABT"", ORTHO-ESTT"", OGENT"" and estropipate (estrone), ESTROVIST""
(quinestrol), ESTRADERMT"" (estradiol), DELESTROGENT"" and VALERGENT""
(estradiol valerate), DEPO-ESTRADIOL CYPIONATET"" and ESTROJECT LAT""
(estradiol cypionate); antiestrogens such as NOLVADEXT"" (tamoxifen), SEROPHENET"" and CLOMIDT"" (clomiphene); progestins such as DURALUTINT""
(hydroxyprogesterone caproate), MPAT"" and DEPO-PROVERAT""
(medroxyprogesterone acetate), PROVERAT"" and CYCRINT"" (MPA), MEGACET""
(megestrol acetate), NORLUTINT"" (norethindrone), and NORLUTATET"" and AYGEST1NT"" (norethindrone acetate); progesterone implants such as NORPLANT
SYSTEMT"" (subdermal implants of norgestrel); antiprogestins such as RU 486T""
(mifepristone); hormonal contraceptives such as ENOVIDT"" (norethynodrel plus mestranol), PROGESTASERTT"" (intrauterine device that releases progesterone), LOESTRINT"', BREVICONT"", MODICONT"', GENORAT"", NELONAT"", NORINYLT"~, OVACON-35T"" and OVACON-SOT"" (ethinyl estradiol/norethindrone), LEVLENT~", NORDETTET"", TRI-LEVLENT"" and TRIPHASIL-21T"" (ethinyl estradiol/levonorgestrel) LO/OVRALT"" and OVRALT"" (ethinyl estradiol/norgestrel), DEMULENT"" (ethinyl estradiol/ethynodiol diacetate), NORINYLT"", ORTHO-NOVUMT"", NORETHINT"", GENORAT"", and NELOVAT"" (norethindrone/mestranol), DESOGENT"" and ORTHO-CEPTT"" (ethinyl estradiol/desogestrel), ORTHO-CYCLENT"" and ORTHO-TRICYCLENT"" (ethinyl estradiol/norgestimate), MICRONORT"" and NOR-QDT"" (norethindrone), and OVRETTET"" (norgestrel).
Additional treatments for endocrine and/or hormone imbalance disorders include, but are not limited to, testosterone esters such as methenolone acetate and testosterone undecanoate; parenteral and oral androgens such as TESTOJECT-SOT""
(testosterone), TESTEXT~" (testosterone propionate), DELATESTRYLT""
(testosterone enanthate), DEPO-TESTOSTERONET"" (testosterone cypionate), DANOCRINET""
(danazol), HALOTESTINT"" (fluoxymesterone), ORETON METHYLT"", TESTREDT""
and VIRILONT"" (methyltestosterone), and OXANDRINT"" (oxandrolone);
testosterone transdermal systems such as TESTODERMT""; androgen receptor antagonist and 5-alpha-reductase inhibitors such as ANDROCURT"" (cyproterone acetate), EULEXINT"" (flutamide), and PROSCART"" (finasteride);
adrenocorticotropic hormone preparations such as CORTROSYNT"" (cosyntropin); adrenocortical steroids and their synthetic analogs such as ACLOVATET"" (alclometasone dipropionate), CYCLOCORTT"" (amcinonide), BECLOVENTT"" and VANCERILT"' (beclomethasone dipropionate), CELESTONET"" (betamethasone), BENISONET"" and UTICORTT""
(betamethasone benzoate), DIPROSONET"" (betamethasone dipropionate), CELESTONE PHOSPHATET"' (betamethasone sodium phosphate), CELESTONE
SOLUSPANT"~ (betamethasone sodium phosphate and acetate), BETA-VALT"" and VALISONET"" (betamethasone valerate), TEMOVATET"" (clobetasol propionate), CLODERMT"" (clocortolone pivalate), CORTEFT"" and HYDROCORTONET""
(cortisol (hydrocortisone)), HYDROCORTONE ACETATET"" (cortisol (hydrocortisone) acetate), LOCOIDT"" (cortisol (hydrocortisone) butyrate), HYDROCORTONE PHOSPHATET"" (cortisol (hydrocortisone) sodium phosphate), A-HYDROCORTT"" and SOLU CORTEFT"" (cortisol (hydrocortisone) sodium succinate), WESTCORTT"" (cortisol (hydrocortisone) valerate), CORTISONE
ACETATET"" (cortisone acetate), DESOWENT"" and TRIDESILONT"" (desonide), TOPICORTT"" (desoximetasone), DECADRONT"" (dexamethasone), DECADRON
LAT"" (dexamethasone acetate), DECADRON PHOSPHATET"" and HEXADROL
PHOSPHATET"" (dexamethasone sodium phosphate), FLORONET"" and MAXIFLORT~" (diflorasone diacetate), FLORINEF ACETATET"" (fludrocortisone acetate), AEROB>DT"" and NASALIDET"" (flunisolide), FLUONIDT"" and SYNALART"" (fluocinolone acetonide), LIDEXT"" (fluocinonide), FLUOR-OPT"" and FMLT"" (fluorometholone), CORDRANT"" (flurandrenolide), HALOGT""
(halcinonide), HMS LIZUIFILMT"" (medrysone), MEDROLT"" (methylprednisolone), DEPO-MEDROLT"" and MEDROL ACETATET"' (methylprednisone acetate), A-METHAPREDT"' and SOLUMEDROLT"" (methylprednisolbne sodium succinate), ELOCONT"" (mometasone furoate), HALDRONET"" (paramethasone acetate), DELTA-CORTEFT"" (prednisolone), ECONOPREDT"" (prednisolone acetate), HYDELTRASOLT"" (prednisolone sodium phosphate), HYDELTRA-T.B.AT""
(prednisolone tebutate), DELTASONET"" (prednisone), ARISTOCORTT"" and KENACORTT"" (triamcinolone), KENALOGT"" (triamcinolone acetonide), ARISTOCORTT"" and KENACORT DIACETATET"" (triamcinolone diacetate), and ARISTOSPANT~" (triamcinolone hexacetonide); inhibitors of biosynthesis and action of adrenocortical steroids such as CYTADRENT"" (aminoglutethimide), NIZORALT""
(ketoconazole), MODRASTANET"" (trilostane), and METOPIRONET"" (metyrapone);
bovine, porcine or human insulin or mixtures thereof; insulin analogs;
recombinant human insulin such as HUMULINT"" and NOVOLINT""; oral hypoglycemic agents such as ORAMIDET"" and ORINASET"" (tolbutamide), DIABINESET""

(chlorpropamide), TOLAMIDET"" and TOLINASET"" (tolazamide), DYMELORT""
(acetohexamide), glibenclamide, MICRONASET"", DIBETAT"" and GLYNASET""
(glyburide), GLUCOTROLT"" (glipizide), and DIAMICRONT"" (gliclazide), GLUCOPHAGET"" (metformin), ciglitazone, pioglitazone, and alpha-glucosidase inhibitors; bovine or porcine glucagon; somatostatins such as SANDOSTATINT""
(octreotide); and diazoxides such as PROGLYCEMT"" (diazoxide).
In one embodiment, the Therapeutics of the invention are administered in combination with treatments for uterine motility disorders. Treatments for uterine motility disorders include, but are not limited to, estrogen drugs such as conjugated estrogens (e.g., PREMARIN° and. ESTRATAB°), estradiols (e.g., CLIMARA° and ALORA°), estropipate; and chlorotrianisene; progestin drugs (e.g., AMEN°
(medroxyprogesterone), MICRONOR° (norethidrone acetate), PROMETRIUM°
progesterone, and megestrol acetate); and estrogen/progesterone combination therapies such as, for example, conjugated estrogens/medroxyprogesterone (e.g., PREMPROT"" and PREMPHASE°) and norethindrone acetate/ethinyl estsradiol (e.g., FEMHRTT"").
In an additional embodiment, the Therapeutics of the invention are administered in combination with drugs effective in treating iron deficiency and hypochromic anemias, including but not limited to, ferrous sulfate (iron sulfate, FEOSOLTM), ferrous fumarate (e.g., FEOSTATTM), ferrous gluconate (e.g., FERGONTM), polysaccharide-iron complex (e.g., NIFEREXTM), iron dextran injection (e.g., INFEDTM), cupric sulfate, pyroxidine, riboflavin, Vitamin B,Z, cyancobalamin injection (e.g., REDISOLTM, RUBRAMIN PCTM), hydroxocobalamin, folic acid (e.g., FOLVITETM), leucovorin (folinic acid, 5-CHOH4PteGlu, citrovorum factor) or WELLCOVORIN (Calcium salt of leucovorin), transferrin or ferritin.
In certain embodiments, the Therapeutics of the invention are administered in combination with agents used to treat psychiatric disorders. Psychiatric drugs that may be administered with the Therapeutics of the invention include, but are not limited to, antipsychotic agents (e.g., chlorpromazine, chlorprothixene, clozapine, fluphenazine, haloperidol, loxapine, mesoridazine, molindone, olanzapine, perphenazine, pimozide, quetiapine, risperidone, thioridazine, thiothixene, trifluoperazine, and triflupromazine), antimanic agents (e.g., carbamazepine, divalproex sodium, lithium carbonate, and lithium citrate), antidepressants (e.g., amitriptyline, amoxapine, bupropion, citalopram, clomipramine, desipramine, doxepin, fluvoxamine, fluoxetine, imipramine, isocarboxazid, maprotiline, 5' mirtazapine, nefazodone, nortriptyline, paroxetine, phenelzine, protriptyline, sertraline, tranylcypromine, trazodone, trimipramine, and venlafaxine), antianxiety agents (e.g., alprazolam, buspirone, chlordiazepoxide, clorazepate, diazepam, halazepam, lorazepam, oxazepam, and prazepam), and stimulants (e.g., d-amphetamine, methylphenidate, and pemoline).
In other embodiments, the Therapeutics of the invention are administered in combination with agents used to treat neurological disorders. Neurological agents that may be administered with the Therapeutics of the invention include, but are not limited to, antiepileptic agents (e.g., carbamazepine, clonazepam, ethosuximide, phenobarbital, phenytoin, primidone, valproic acid, divalproex sodium, felbamate, gabapentin, lamotrigine, levetiracetam, oxcarbazepine, tiagabine, topiramate, zonisamide, diazepam, lorazepam, and clonazepam), antiparkinsonian agents (e.g:, levodopa/carbidopa, selegiline, amantidine, bromocriptine, pergolide, ropinirole, pramipexole, benztropine; biperiden; ethopropazine; procyclidine;
trihexyphenidyl, tolcapone), and ALS therapeutics (e.g. riluzole).
In another embodiment, Therapeutics of the invention are administered in combination with vasodilating agents and/or calcium channel blocking agents.
Vasodilating agents that may be administered with the Therapeutics of the invention include, but are not limited to, Angiotensin Converting Enzyme (ACE) inhibitors (e.g., papaverine, isoxsuprine, benazepril, captopril, cilazapril, enalapril, enalaprilat, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, spirapril, trandolapril, and nylidrin), and nitrates (e.g., isosorbide dinitrate, isosorbide mononitrate, and nitroglycerin). Examples of calcium channel blocking agents that may be administered in combination with the Therapeutics of the invention include, but are not limited to amlodipine, bepridil, diltiazem, felodipine, flunarizine, isradipine, nicardipine, nifedipine, nimodipine, and verapamil.

In additional embodiments, the Therapeutics of the invention are administered in combination with other therapeutic or prophylactic regimens, such as, for example, radiation therapy.
Example 24: Method of Treating Decreased Levels of the Polypeptide The present invention relates to a method for treating an individual in need of an increased level of a polypeptide of the invention in the body comprising administering to such an individual a composition comprising a therapeutically effective amount of an agonist of the invention (including polypeptides of the invention). Moreover, it will be appreciated that conditions caused by a decrease in the standard or normal expression level of a secreted protein in an individual can be treated by administering the polypeptide of the present invention, preferably in the secreted form. Thus, the invention also provides a method of treatment of an individual in need of an increased level of the polypeptide comprising administering to such an individual a Therapeutic comprising an amount of the polypeptide to increase the activity level of the polypeptide in such an individual.
For example, a patient with decreased levels of a polypeptide receives a daily dose 0.1-100 ug/kg of the polypeptide for six consecutive days. Preferably, the polypeptide is in the secreted form. The exact details of the dosing scheme, based on .
administration and formulation, are provided in Example 23.
Example 25: Method of Treating Increased Levels of the Polypeptide The present invention also relates to a method of treating an individual in need of a decreased level of a polypeptide of the invention in the body comprising administering to such an individual a composition comprising a therapeutically effective amount of an antagonist of the invention (including polypeptides and antibodies of the invention).
In one example, antisense technology is used to inhibit production of a polypeptide of the present invention. This technology is one example of a method of decreasing levels of a polypeptide, preferably a secreted form, due to a variety of etiologies, such as cancer. For example, a patient diagnosed with abnormally increased levels of a polypeptide is administered intravenously antisense polynucleotides at 0.5, 1.0, 1.5, 2.0 and 3.0 mg/kg day for 21 days. This treatment is repeated after a 7-day rest period if the treatment was well tolerated. The formulation of the antisense polynucleotide is provided in Example 23.
Example 26:' Method of Treatment Using Gene Therapy-Ex Vivo One method of gene therapy transplants fibroblasts, which are capable of expressing a polypeptide, onto a patient. Generally, fibroblasts are obtained from a subject by skin biopsy. The resulting tissue is placed in tissue-culture medium and separated into small pieces. Small chunks of the tissue are placed on a wet surface of a tissue culture flask, approximately ten pieces are placed in each flask. The flask is turned upside down, closed tight and left at room temperature over night.
After 24 hours at room temperature, the flask is inverted and the chunks of tissue remain fixed to the bottom of the flask and fresh media (e.g., Ham's F12 media, with 10%
FBS, penicillin and streptomycin) is added. The flasks are then incubated at 37 degree C
for approximately one week.
At this time, fresh media is added and subsequently changed every several days: After an additional two weeks in culture, a monolayer of fibroblasts emerge.
The monolayer is trypsinized and scaled into larger flasks.
pMV-7 (Kirschmeier, P.T. et al., DNA, 7:219-25 (1988)), flanked by the long terminal repeats of the Moloney murine sarcoma virus, is digested with EcoRI
and HindIII and subsequently treated with calf intestinal phosphatase. The linear vector is fractionated on agarose gel and purified, using glass beads.
The cDNA encoding a polypeptide of the present invention can be amplified using PCR primers which correspond to the 5' and 3' end sequences respectively as set forth in Example 1 using primers and having appropriate restriction sites and initiation/stop codons, if necessary. Preferably, the 5' primer contains an EcoRI site and the 3' primer includes a HindIII'site. Equal quantities of the Moloney murine sarcoma virus linear backbone and the amplified EcoRI and HindIII fragment are added together, in the presence of T4 DNA ligase. The resulting mixture is maintained under conditions appropriate for ligation of the two fragments. The ligation mixture is then used to transform bacteria HB101; which are then plated onto agar containing kanamycin for the purpose of confirming that the vector has the gene of interest properly inserted. .
The amphotropic pA317 or GP+arnl2 packaging cells are grown in tissue culture to confluent density in Dulbecco's Modified Eagles Medium (DMEM) with 10% calf serum (CS), penicillin and streptomycin. The MSV vector containing the gene is then added to the media and the packaging cells transduced with the vector.
The packaging cells now produce infectious viral particles containing the gene (the packaging cells are now referred to as producer cells).
Fresh media is added to the transduced producer cells, and subsequently, the media is harvested from a 10 cm plate of confluent producer cells. The spent media, containing the infectious viral particles, is filtered through a millipore filter to remove detached producer cells and this media is then used to infect fibroblast cells. Media is removed from a sub-confluent plate of fibroblasts and quickly replaced with the media from the producer cells. This media is removed and replaced with fresh media.
If the titer of virus is high, then virtually all fibroblasts will be infected and no selection is required. If the titer is very low, then it is necessary to use a retroviral vector that has a selectable marker, such as neo or his. Once the fibroblasts have been efficiently infected, the fibroblasts are analyzed to determine whether protein is produced.
The engineered fibroblasts are then transplanted onto the host, either alone or after having been grown to confluence on cytodex 3 microcarrier beads.
Example 27: Gene Therapy Using Endogenous Genes Corresponding To Polynucleotides of the Invention Another method of gene therapy according to the present invention involves operably associating the endogenous polynucleotide sequence of the invention with a promoter via homologous recombination as described, for example, in U.S.
Patent NO: 5,641,670, issued June 24, 1997; International Publication NO: WO
96/29411, published September 26, 1996; International Publication NO: WO 94/12650, published August 4, 1994; Koller et al., Proc. Natl. Acad. Sci. USA, 86:8932-8935, (1989); and Zijlstra et al., Nature, 342:435-438 (1989). This method involves the activation of.a gene which is present in the target cells, but which is not expressed in the cells, or is expressed at a lower level than desired.
Polynucleotide constructs are made which contain a promoter and targeting sequences, which are homologous to the 5' non-coding sequence of endogenous polynucleotide sequence, flanking the promoter. The targeting sequence will be sufficiently near the 5' end of the polynucleotide sequence so the promoter will be operably linked to the endogenous sequence upon homologous recombination. The promoter and the targeting sequences can be amplified using PCR. Preferably, the amplified promoter contains distinct restriction enzyme sites on the 5' and 3' ends.
Preferably, the 3' end of the first targeting sequence contains the same restriction enzyme site as the 5' end of the amplified promoter and the 5' end of the second targeting sequence contains the same restriction site as the 3' end of the amplified promoter.
The amplified promoter and the amplified targeting sequences are digested with the appropriate restriction enzymes and subsequently treated with calf intestinal phosphatase. The digested promoter and digested targeting sequences are added together in the presence of T4 DNA lipase. The resulting mixture is maintained under conditions appropriate for ligation of the two fragments. The construct is size fractionated on an agarose gel then purified by phenol extraction and ethanol precipitation.
In this Example, the polynucleotide constructs are administered as naked polynucleotides via electroporation. However, the polynucleotide constructs may also be administered with transfection-facilitating agents, such as liposomes, viral sequences, viral particles, precipitating agents, etc. Such methods of delivery are known in the art.
Once the cells are transfected, homologous recombination will take place which results in the promoter being operably linked to the endogenous polynucleotide sequence. This results in the expression of polynucleotide corresponding to the polynucleotide in the cell. Expression may be detected by immunological staining, or any other method known in the art.
Fibroblasts are obtained from a subject by skin biopsy. The resulting tissue is placed in DMEM + 10% fetal calf serum. Exponentially growing or early stationary phase fibroblasts are trypsinized and rinsed from the plastic surface with nutrient medium. An aliquot of the cell suspension is removed for counting, and the remaining cells are subjected to centrifugation. The supernatant is aspirated and the pellet is resuspended in 5 ml of electroporation buffer (20 mM HEPES pH 7.3, 137 mM
NaCI, 5 mM KC1, 0.7 mM Na2 HP04, 6 mM dextrose). The cells are recentrifuged, the supernatant aspirated, and the cells resuspended in electroporation buffer containing 1 mg/ml acetylated bovine serum albumin. The final cell suspension contains, approximately 3X106 cells/ml: Electroporation should be performed immediately following resuspension.
Plasmid DNA is prepared according to standard techniques. For example, to construct a plasmid for targeting to the locus corresponding to the polynucleotide of the invention, plasmid pUCl8 (MBI Fermentas, Amherst, NY) is digested with HindIII. The CMV promoter is amplified by PCR with an XbaI site on the 5' end and a BamHI site on the 3'end. Two non-coding sequences are amplified via PCR: one non-coding sequence (fragment 1 ) is amplified with a HindIII site at the 5' end and an Xba site at the 3'end; the other non-coding sequence (fragment 2) is amplified with a BamHI site at the 5'end and a HindIII site at the 3'end. The CMV promotei and the fragments (1 and 2) are digested with the appropriate enzymes (CMV promoter -XbaI
and BamHI; fragment 1 - XbaI; fragment 2 - BamHI) and ligated together. The resulting ligation product is digested with HindIII, and ligated with the HindIII-digested pUCl8 plasmid.
Plasmid DNA is added to a sterile cuvette with a 0.4 cm electrode gap (Bio-Rad). The final DNA concentration is generally at least 120 ~g/ml. 0.5 ml of the cell suspension (containing approximately 1.5.X106 cells) is then added to the cuvette, and the cell suspension and DNA solutions are gently mixed. Electroporation is performed with a Gene-Pulser apparatus (Bio-Rad). Capacitance and voltage are set at 960 pF and 250-300 V, respectively. As voltage increases, cell survival decreases, but the percentage of surviving cells that stably incorporate the introduced DNA
into their genome increases dramatically. Given these parameters, a pulse time of approximately 14-20 mSec should be observed.
Electroporated cells are maintained at room temperature for approximately 5 min, and the contents of the cuvette are then gently removed with a sterile transfer pipette. The cells are added directly to 10 ml of prewarmed nutrient media (DMEM
with 15% calf serum) in a 10 cm dish and incubated at 37 degree C. The following day, the media is aspirated and replaced with 10 ml of fresh media and incubated for a further 16-24 hours.
The engineered fibroblasts are then injected into the host, either alone or after having been grown to confluence on cytodex 3 microcarrier beads. The fibroblasts now produce the protein product. The fibroblasts can then be introduced into a patient as described above.
Example 28: Method of Treatment Using Gene Therapy - In Vivo Another aspect of the present invention is using in vivo gene therapy methods to treat disorders, diseases and conditions. The gene therapy method relates to the introduction of naked nucleic acid (DNA, RNA, and antisense DNA or RNA) sequences into an animal to increase or decrease the expression of the polypeptide.
The polynucleotide of the present invention may be operatively linked to a promoter or any other genetic elements necessary for the expression of the polypeptide by the target tissue. Such gene therapy and delivery techniques and methods are known in the art, see, for example, W090/11092, W098/11779; U.S. Patent NO. 5693622, 5705151, 5580859; Tabata et al., Cardiovasc. Res. 35(3):470-479 (1997); Chao et al., Pharmacol. Res. 35(6):517-522 (1997); Wolff, Neuromuscul. Disord. 7(5):314-318 (1997); Schwartz et al., Gene Ther. 3(5):405-411 (1996); Tsurumi et al., Circulation 94(12):3281-3290 (1996) (incorporated herein by reference).
The polynucleotide constructs may be delivered by any method that delivers injectable materials to the cells of an animal, such as, injection into the interstitial space of tissues (heart, muscle, skin, lung, liver, intestine and the like).
The polynucleotide constructs can be delivered in a pharmaceutically acceptable liquid or aqueous carrier.
The term "naked" polynucleotide, DNA or RNA, refers to sequences that are free from any delivery vehicle that acts to assist, promote, or facilitate entry into the cell, including viral sequences, viral particles, liposome formulations, lipofectin or precipitating agents and the like. However, the polynucleotides of the present .
invention may also be delivered in liposome formulations (such as those taught in _.

Felgner P.L. et al. (1995) Ann. NY Acad. Sci. 772:126-139 and Abdallah B. et al.
(1995) Biol. Cell 85(1):1-7) which can be prepared by methods well known to those skilled in the art.
The polynucleotide vector constructs used in the gene therapy method are preferably constructs that will not integrate into the host genome nor will they contain sequences that allow for replication. Any strong promoter known to those skilled in the art can be used for driving the expression of DNA. Unlike other gene therapies techniques, one major advantage of introducing naked nucleic acid sequences into target cells is the transitory nature of the polynucleotide synthesis in the cells. Studies have shown that non-replicating DNA sequences can be introduced into cells to provide production of the desired polypeptide for periods of up to six months.
The polynucleotide construct can be delivered to the interstitial space of tissues within the an animal, including of muscle, skin, brain, lung, liver, spleen, bone marrow, thymus, heart, lymph, blood, bone, cartilage, pancreas, kidney, gall bladder, stomach, intestine, testis, ovary, uterus, rectum, nervous system, eye, gland, and connective tissue. Interstitial space of the tissues comprises the intercellular fluid, mucopolysaccharide matrix among the reticular fibers of organ tissues, elastic fibers in the walls of vessels or chambers, collagen fibers of fibrous tissues, or that same matrix within connective tissue ensheathing muscle cells or in the lacunae of bone. It is similarly the space occupied by the plasma of the circulation and the lymph fluid of the lymphatic channels. Delivery to the interstitial space of muscle tissue is preferred for the reasons discussed below. They may be conveniently delivered by injection into the tissues comprising these cells. They are preferably delivered to and expressed in persistent, non-dividing cells which are differentiated, although delivery and expression maybe achieved in non-differentiated or less completely differentiated cells, such as, for example, stem cells of blood or skin fibroblasts. In vivo muscle cells are particularly competent in their ability to take up and express polynucleotides.
For the naked polynucleotide injection, an effective dosage amount of DNA or RNA will be in the range of from about 0.05 g/kg body weight to about 50 mg/kg body weight. Preferably the dosage will be from about 0.005 mg/kg to about 20 mg/kg and more preferably from about 0.05 mg/kg to about 5 mg/kg. Of course, as the artisan of ordinary skill will appreciate, this dosage will vary according to the tissue site of injection. The appropriate and effective dosage of nucleic acid sequence can readily be determined by those of ordinary skill in the art and may depend on the condition being treated and the route of administration. The preferred route of administration is by the parenteral route of injection into the interstitial space of tissues. However, other parenteral routes may also be usef, such as, inhalation of an aerosol formulation particularly for delivery to lungs or bronchial tissues, throat or mucous membranes of the nose. In addition, naked polynucleotide constructs can be delivered to arteries during angioplasty by the catheter used ~in the procedure.
The dose response effects of injected polynucleotide in muscle in vivo is determined as follows. Suitable template DNA for production of mRNA coding for polypeptide of the present invention is prepared in accordance with a standard recombinant DNA methodology. The template DNA, which may be either circular or linear, is either used as naked DNA or complexed with liposomes. The quadriceps muscles of mice are then injected with various amounts of the template DNA.
Five to six week old female and male Balb/C mice are anesthetized by intraperitoneal injection with 0.3 ml of 2.5% Avertin. A 1.5 cm incision is made on the anterior thigh, and the quadriceps muscle is directly visualized. The template DNA is injected in 0.1 ml of carrier in a 1 cc syringe through a 27 gauge needle over one minute, approximately 0.5 cm from the distal insertion site of the muscle into the . knee and about 0.2 cm deep. A suture is placed over the injection site for future localization, and the skin is closed with, stainless steel clips.
After an appropriate incubation time (e.g., 7 days) muscle extracts are prepared by excising the entire quadriceps. Every fifth 15 um cross-section of the individual quadriceps muscles is histochemically stained for protein expression. A
time course for protein expression may be done in a similar fashion except that quadriceps from different mice are harvested at different times. Persistence of DNA
in muscle following injection may be determined by Southern blot analysis after preparing total cellular DNA and HIRT supernatants from injected and control mice.
The results of the above experimentation in mice can be use to extrapolate proper dosages and other treatment parameters in humans and other animals using naked DNA.

Example 29: Trans~enic Animals.
The polypeptides of the invention can also be expressed in transgenic animals.
Animals of any species, including, but not limited to, mice, rats, rabbits, hamsters, guinea pigs, pigs, micro-pigs, goats, sheep, cows and non-human primates, e.g., baboons, monkeys, and chimpanzees may be used to generate transgenic animals.
In a specific embodiment, techniques described herein or otherwise known in the art, are used to express polypeptides of the invention in humans, as part of a gene therapy protocol.
Any technique known in the art may be used to introduce the transgene (i.e., polynucleotides of the invention) into animals to produce the founder lines of transgenic animals. Such techniques include, but are not limited to, pronuclear microinjection (Paterson et al., Appl. Microbiol. Biotechnol. 40:691-698 (1994);
Carver et al., Biotechnology (NY) 11:1263-1270 (1993); Wright et al., Biotechnology (NY) 9:830-834 (1991); and Hoppe et al., U.S. Pat. No. 4,873,191 (1989));
retrovirus mediated gene transfer into germ lines (Van der Putten et al., Proc. Natl.
Acad. Sci., USA 82:6148-6152 (1985)), blastocysts or embryos; gene targeting in embryonic stem cells (Thompson et al., Cell 56:313-321 ( 1989)); electroporation of cells or embryos (Lo, 1983, Mol Cell. Biol. 3:1803-1814 (1983)); introduction of the polynucleotides of the invention using a gene gun (see, e.g., Ulmer et al., Science 259:1745 (1993); introducing nucleic acid constructs into embryonic pleuripotent stem cells and transferring the stem cells back into the blastocyst; and sperm-mediated gene transfer (Lavitrano et al., Cell 57:717-723 (1989); etc. For a review of such techniques, see Gordon, "Transgenic Animals," Intl. Rev. Cytol. 115:171-(1989), which is incorporated by reference herein in its entirety.
Any technique known in the art may be used to produce transgenic clones containing polynucleotides of the invention, for example, nuclear transfer into enucleated oocytes of nuclei from cultured embryonic, fetal, or adult cells induced to quiescence (Campell et al., Nature 380:64-66 (1996); Wilmut et al., Nature 385:810-813 (1997)).
The present invention provides for transgenic animals that carry the transgene in all their cells, as well as animals which carry the transgene in some, but not all their cells, i.e., mosaic animals or chimeric. The transgene may be integrated as a single transgene or as multiple copies such as in concatamers, e.g., head-to-head tandems or head-to-tail tandems. The transgene may also be selectively introduced into and activated in a particular cell type by following, for example, the teaching of Lasko et al. (Lasko et al., Proc. Natl. Acad. Sci. USA 89:6232-6236 (1992)). The regulatory sequences required for such a cell-type specific activation will depend upon the particular cell type of interest, and will be apparent to those of skill in the art. When it is desired that the polynucleotide transgene be integrated into the chromosomal site of the endogenous gene, gene targeting is preferred. Briefly, when such a technique is to be utilized, vectors containing some nucleotide sequences homologous to the endogenous gene are designed for the purpose of integrating, via homologous recombination with chromosomal sequences, into and disrupting the function of the nucleotide sequence of the endogenous gene. The transgene may also be selectively introduced into a particular cell type, thus inactivating the endogenous gene in only that cell type, by following, for example, the teaching of Gu et al. (Gu et al., Science 265:103-106 (1994)). The regulatory sequences required for such a cell-type specific inactivation will depend upon the particular cell type of interest, and will be apparent to those of skill in the art.
Once transgenic animals have been generated, the expression of the recombinant gene may be assayed utilizing standard techniques. Initial screening may be accomplished by Southern blot analysis or PCR techniques to analyze animal tissues to verify that integration of the transgene has taken place. The level of mRNA
expression of the transgene iri the tissues of the transgenic animals may also be assessed using techniques which include, but are not limited to, Northern blot analysis of tissue samples obtained from the animal, in situ hybridization analysis, .and reverse transcriptase-PCR (rt-PCR). Samples of transgenic gene-expressing tissue may also be evaluated immunocytochemically or immunohistochemically using antibodies specific for the transgene product.
Once the founder animals are produced, they may be bred, inbred, outbred, or 30- crossbred to produce colonies of the particular animal. Examples of such breeding strategies include, but are not limited to: outbreeding of founder animals with more than one integration site in order to establish separate lines; inbreeding of separate lines in order to produce compound transgenics that express the transgene at higher levels because of the effects of additive expression of each transgene;
crossing of heterozygous transgenic animals to produce animals homozygous for a given integration site in order to both augment expression and eliminate the need for screening of animals by DNA analysis; crossing of separate homozygous lines to produce compound heterozygous or homozygous lines; and breeding to place the transgene on a distinct background that is appropriate for an experimental model of interest.
Transgenic animals of the invention have uses which include, but are not limited to, animal model systems useful in elaborating the biological function of polypeptides of the present invention, studying diseases, disorders, and/or conditions associated with aberrant expression, and in screening for compounds effective in ameliorating such diseases, disorders, and/or conditions.
Examule 30: Knock-Out Animals.
Endogenous gene expression can also be reduced by inactivating or "knocking out" the gene and/or its promoter using targeted homologous recombination.
(E.g., see Smithies et al., Nature 317:230-234 (1985); Thomas & Capecchi, Cell 51:503-512 (1987); Thompson et al., Cell 5:313-321 (1989); each of which is incorporated by reference herein in its entirety). For example, a mutant, non-functional polynucleotide of the invention (or a completely unrelated DNA sequence) flanked by DNA homologous to the endogenous polynucleotide sequence (either the coding regions or regulatory regions of the gene) can be used, with or without a selectable marker and/or a negative selectable marker, to transfect cells that express polypeptides of the invention in vivo. In another embodiment, techniques known in the art are used to generate knockouts in cells that contain, but do not express the gene of interest. Insertion of the DNA construct, via targeted homologous recombination, results in inactivation of the targeted gene. Such approaches are particularly suited in research and agricultural fields where modifications to embryonic stem cells can be used to generate animal offspring with an inactive targeted gene (e.g., see Thomas &
Capecchi 1987 and Thompson 1989, supra). However this approach can be routinely adapted for use in humans provided the recombinant DNA constructs are directly administered or targeted to the required site in vivo using appropriate viral vectors that will be apparent to those of skill in the art.
In further embodiments of the invention, cells that are genetically engineered to express the polypeptides of the invention, or alternatively, that are genetically engineered not fo express the polypeptides of the invention (e.g., knockouts) are administered to a patient in vivo. Such cells may be obtained from the patient (i.e., animal, including human) or an MHC compatible donor and can include, but are not limited to fibroblasts, bone marrow cells, blood cells (e.~, lymphocytes), adipocytes, muscle cells, endothelial cells etc. The cells are genetically engineered in vitro using recombinant DNA techniques to introduce the coding sequence of polypeptides of the invention into the cells, or alternatively, to disrupt the coding sequence and/or endogenous regulatory sequence associated with the polypeptides of the invention, e.~, by transduction (using viral vectors, and preferably vectors that integrate the transgene into the cell genome) or transfection procedures, including, but not limited to, the use of plasmids, cosmids, YACs, naked DNA, electroporation, liposomes, etc.
The coding sequence of the polypeptides of the invention can.be placed under the .
control of a strong constitutive or inducible promoter or prorrioter/enhancer to achieve expression, and preferably secretion, of the polypeptides~of the invention.
The engineered cells which express and preferably secrete the polypeptides of the invention can be introduced into the patient systemically, e.g., in the circulation, or intraperitoneally..
Alternatively, the cells can be incorporated into a matrix and implanted in the body, ~, genetically engineered fibroblasts can be implanted as part of a skin graft;
genetically engineered endothelial cells can be implanted as part of a lymphatic or vascular graft. (See, for example, Anderson et al. U.S. Patent No. 5,399,349;
and Mulligan & Wilson, U.S. Patent No. 5,460,959 each of which is incorporated by reference herein in its entirety).
When the cells to be administered are non-autologous or non-MHC
compatible cells, they can be administeredwsing well known techniques which prevent the development of a host immune response against the introduced cells. For example, the cells may be introduced in an encapsulated form which, while allowing for an exchange of components with the immediate extracellular environment, does not allow the introduced cells to be recognized by the host immune system.
Transgenic and "knock-out" animals of the invention have uses which include, but are not limited to, animal model systems useful in elaborating the biological function of polypeptides of the present invention, studying diseases, disorders, and/or conditions associated with aberrant expression, and in screening for compounds effective in ameliorating such diseases, disorders, and/or conditions.
Example 31: Production of an Antibody Hybridoma Technology The antibodies of the present invention can be prepared by a variety of methods. (See, Current Protocols, Chapter 2.) As one example of such methods, cells expressing polypeptide(s) of the invention are administered to an animal to induce the production of sera containing polyclonal antibodies. In a preferred method, a preparation of polypeptide(s) of the invention is prepared and purified to render it substantially free of natural contaminants. Such a preparation is then introduced into an animal in order to produce polyclonal antisera of greater specific activity.
Monoclonal antibodies specific for polypeptide(s) of the invention are prepared using hybridoma technology. (Kohler et al., Nature 256:495 (1975);
Kohler et al., Eur. J. Immunol. 6:511 (1976); Kohler et al., Eur. J. Immunol. 6:292 (1976);
Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas, Elsevier, N.Y., pp. 563-681 (1981)). In general, an animal (preferably a mouse) is immunized with polypeptide(s) of the invention, or, more preferably, with a secreted polypeptide-expressing cell. Such polypeptide-expressing cells are cultured in any suitable tissue culture medium, preferably in Earle's modified Eagle's medium supplemented with 10% fetal bovine serum (inactivated at about 56°C), and supplemented with about 10 g/I of nonessential amino acids, about 1,000 U/ml of penicillin, and about 100 ~g/ml of streptomycin.
The splenocytes of such mice are extracted and fused with a suitable myeloma cell line. Any suitable myeloma cell line may be employed in accordance with the present invention; however, it is preferable to employ the parent myeloma cell line (SP20), available from the ATCC. After fusion, the resulting hybridoma cells are ' _, selectively maintained in HAT medium, and then cloned by limiting dilution as described by Wands et al. (Gastroenterology 80:225-232 (1981)). The hybridoma cells obtained through such a selection are then assayed to identify clones which secrete antibodies capable of binding the polypeptide(s) of the invention.
Alternatively, additional antibodies capable of binding polypeptide(s) of the invention can be produced in a two-step procedure using anti-idiotypic antibodies.
Such a method makes use of the fact that antibodies are themselves antigens, and therefore, it is possible to obtain an antibody which binds to a second antibody. In accordance with this method, protein specific antibodies are used to immunize an animal, preferably a mouse. The splenocytes of such an animal are then used to produce hybridoma cells, and the hybridoma cells are screened to identify clones which produce an antibody whose ability to bind to the polypeptide(s) of the invention protein-specific antibody can be blocked by polypeptide(s) of the invention.
Such antibodies comprise anti-idiotypic antibodies to the polypeptide(s) of the invention protein-specific antibody and are used to immunize an animal to induce formation of further polypeptide(s),of the invention protein-specific antibodies.
For in vivo use of antibodies in humans, an antibody is "humanized". Such antibodies can be produced using genetic constructs derived from hybridoma cells producing the monoclonal antibodies described above. Methods for producing chimeric and humanized antibodies are known in the art and are discussed herein.
(See, for review, Mornson, Science 229:1202 ( 1985); Oi et al., BioTechniques 4:214 (1986); Cabilly et al., U.S. Patent No. 4,816,567; Taniguchi et al., EP
171496;
Mornson et al., EP 173494; Neuberger et al:, WO 8601533; Robinson et al., WO
8702671; Boulianne et al., Nature 312:643 (1984); Neuberger et,al., Nature 314:268 (1985).) Isolation Of Antibody Fragments Directed polypeptide(s) of the invention From A
Library Of scFvs Naturally occurring V-genes isolated from human PBLs are constructed into a library of antibody fragments which contain reactivities against polypeptide(s) of the invention to which the donor may or may not have been exposed (see e.g., U.S.
Patent 5,885,793 incorporated herein by reference in its entirety).

Rescue of the Library. A library of scFvs is constructed from the RNA of human PBLs as described in PCT publication WO 92/01047. To rescue phage displaying antibody fragments, approximately 109 E. coli harboring the phagemid are used to inoculate 50 ml of 2xTY containing 1% glucose and 100 p.g/ml of ampicillin (2xTY-AMP-GLU) and grown to an O.D. of 0.8 with shaking. Five ml of this culture is used to irmoculate 50 ml of 2xTY-AMP-GLU, 2 x 108 TU of delta gene 3 helper (M13 delta gene III, see PCT publication WO 92/01047) are added and the culture incubated at 37°C for 45 minutes without shaking and then at 37°C for 45 minutes with shaking. The culture is centrifuged at 4000 r.p.m. for 10 min. and the pellet resuspended in 2 liters of 2xTY containing 100 ~ug/ml ampicillin and 50 ug/ml kanamycin and grown overnight. Phage are prepared as described in PCT
publication WO 92/01047.
M 13 delta gene III is prepared as follows: M 13 delta gene III helper phage does not encode gene III protein, hence the phage(mid) displaying antibody fragments have a greater avidity of binding to antigen. Infectious M13 delta gene III
particles are made by growing the helper phage in cells harboring a pUCl9 derivative supplying the wild type gene III protein during phage motphogenesis. The culture is incubated for 1 hour at 37° C without shaking and then for a further hour at 37°C with shaking. Cells are spun down (IEC-Centra 8,400 r.p.m. for 10 min), resuspended in 300 ml 2xTY broth containing 100 pg ampicillin/ml and 25 pg kanamycin/ml (2xTY-AMP-KAN) and grown overnight, shaking at 37°C. Phage particles are purified and concentrated from the culture medium by two PEG-precipitations (Sambrook et al., 1990), resuspended in 2 ml PBS and passed through a 0.45 pm filter (Minisart NML;
Sartorius) to give a final concentration of approximately 1013 transducing units/ml (ampicillin-resistant clones).
Panning of the Library. Immunotubes (Nunc) are coated overnight in PBS
with 4 ml of either 100 p.g/ml or 10 pg/ml of a polypeptide of the present invention.
Tubes are blocked with 2% Marvel-PBS for 2 hours at 37°C and then washed 3'times in PBS. Approximately 1013 TU of phage is applied to the tube and incubated for 30 minutes at room temperature tumbling on an over and under turntable and then left to stand for another 1.5 hours. Tubes are washed 10 times with PBS 0.1% Tween-20 and 10 times with PBS. Phage are eluted by adding 1 ml of 100 mM triethylamine and ..

rotating 15 minutes on an under and over turntable after which the solution is immediately neutralized with 0.5 ml of 1.0M Tris-HCI, pH 7.4. Phage are then used to infect 10 ml of mid-log E. coli TG1 by incubating eluted phage with bacteria for 30 minutes at 37°C. The E. coli are then plated on TYE plates containing 1% glucose and 100 pg/ml ampicillin. The resulting bacterial library is then rescued with delta gene 3 helper phage as described above to prepare phage for a subsequent round of selection. This process is then repeated for a total of 4 rounds of affinity purification , with tube-washing increased to 20 times with PBS, 0.1 % Tween-20 and 20 times with PBS for rounds 3 and 4.
Characterization of Binders. Eluted phage from the 3rd and 4th rounds of selection are used to infect E. coli HB 2151 and soluble scFv is produced (Marks, et al., 1991) from single colonies for assay. ELISAs are performed with microtitre plates coated with either 10 pg/ml of the polypeptide of the present invention in 50 mM bicarbonate pH 9.6. Clones positive in ELISA are further characterized by PCR
fingerprinting (see, e.g., PCT publication WO 92/01047) and then by sequencing.
These ELISA positive clones may also be further characterized by techniques known in the art, such as, for example, epitope mapping, binding affinity, receptor signal transduction, ability to block or competitively inhibit antibody/antigen binding, and competitive agonistic or antagonistic activity.
Example 32: Assays Detecting Stimulation or Inhibition of B cell Proliferation and Differentiation Generation of functional humoral immune responses requires both soluble and cognate signaling between B-lineage cells and their microenvironment. Signals may impart a positive stimulus that allows a B-lineage cell to continue its programmed development, or a negative stimulus that instructs the cell to arrest its current developmental pathway. To date, numerous stimulatory and inhibitory signals have been found to influence B cell responsiveness including IL-2, IL-4, IL-5, IL-6, IL-7, IL10, IL-13,~IL-14 and IL-15. Interestingly, these signals are by themselves weak effectors but can, in combination with various co-stimulatory proteins, induce activation, proliferation, differentiation, homing, tolerance and death among B cell populations.

One of the best studied classes of B-cell co-stimulatory proteins is the TNF-superfamily. Within this family CD40, CD27, and CD30 along with their respective ligands CD154, CD70, and CD153 have been found to regulate a variety of immune responses. Assays which allow for the detection and/or observation of the proliferation and differentiation of these B-cell populations and their precursors are valuable tools in determining the effects various proteins may have on these B-cell populations in terms of proliferation and differentiation. Listed below are two assays designed to allow for the detection of the differentiation, proliferation, or inhibition of B-cell populations and their precursors.
In Vitro Assay- Purified polypeptides of the invention, or truncated forms thereof, is assessed for its ability to induce activation, proliferation, differentiation or inhibition and/or death in B-cell populations and their precursors. The activity of the polypeptides of the invention on purified human tonsillar B cells, measured qualitatively over the dose range from 0.1 to 10,000 ng/mL, is assessed in a standard B-lymphocyte co-stimulation assay in which purified tonsillar B cells are cultured in the presence of either formalin-fixed Staphylococcus aureus Cowan I (SAC) or immobilized anti-human IgM antibody as the priming agent. Second signals such as IL-2 and IL-15 synergize with SAC and IgM crosslinking to elicit B cell proliferation as measured by tritiated-thymidine incorporation. Novel synergizing agents can be readily identified using this assay. The assay involves isolating human tonsillar B
cells by magnetic bead (MACS) depletion of CD3-positive cells. The resulting cell population is greater than 95% B cells as assessed by expression of CD45R(B220).
Various dilutions of each sample are placed into individual wells of a 96-well plate to which are added 105 B-cells suspended in culture medium (RPMI 1640 containing 10%
FBS, 5 X 10-5M 2ME, 100U/ml penicillin, l0ug/ml streptomycin, and 10-5 dilution of SAC) in a total volume of 150u1. Proliferation or inhibition is quantitated by a 20h pulse (luCi/well) with 3H-thymidine (6.7 Ci/mM) beginning 72h post factor addition.
The positive and negative controls are IL2 and medium respectively.
In Vivo Assay- BALB/c mice are injected (i.p.) twice per day with buffer only, or 2 mg/Kg of a polypeptide of the invention, or truncated forms thereof. Mice receive this treatment for 4 consecutive days, at which time they are sacrificed and various tissues and serum collected for analyses. Comparison of H&E sections from normal spleens and spleens treated with polypeptides of the invention identify the results of the activity o.f the polypeptides on spleen cells, such as the diffusion of peri-arterial lymphatic sheaths, and/or significant increases in the nucleated cellularity of the red pulp regions, which may indicate the activation of the differentiation and proliferation of B-cell populations. Immunohistochemical studies using a B
cell marker, anti-CD45R(B220), are used to determine whether any physiological changes to splenic cells, such as splenic disorganization, are due to increased B-cell representation within loosely defined B-cell zones that infiltrate established T-cell regions.
~ Flow cytometric analyses of the spleens from mice treated with polypeptide is used to indicate whether the polypeptide specifically increases the proportion of ThB+, CD45R(B220)dull B cells over that which is observed in control mice.
Likewise, a predicted consequence of increased mature B-cell representation in vivo is a relative increase in serum Ig titers. Accordingly, serum IgM and IgA
levels are compared between buffer and polypeptide-treated mice.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides of the invention (e.g., gene therapy), agonists, and/or antagonists of polynucleotides or polypeptides of the invention.
Example 33: T Cell Proliferation Assay Proliferation assay for Resting PBLs.
A CD3-induced proliferation assay is performed on PBMCs and is measured by the uptake of 3H-thymidine. The assay is performed as follows. Ninety-six well plates are coated with 100 microliters per well of mAb to CD3 (HIT3a, Pharmingen) or isotype-matched control mAb (B33.1) overnight at 4 C (1 microgram/ml in .05M bicarbonate buffer, pH
9.5), then washed three times with PBS. PBMC are isolated by F/H gradient centrifugation from human peripheral blood and added to quadruplicate wells (5 x 104/well) of mAb coated plates in RPMI containing 10% FCS and P/S in the presence of varying concentrations~of TNF Delta and/or TNF Epsilon protein (total volume 200 microliters). Relevant protein buffer and medium alone are controls.
After 48 hr. culture at 37 C, plates are spun for 2 min. at 1000 rpm and 100 microliters of supernatant is removed and stored -20 C for measurement of IL-2 (or other cytokines) if effect on proliferation is observed. Wells are supplemented with 100 microliters of medium containing 0.5 microcuries of 3H-thymidine and cultured at 37 C for 18-24 hr. Wells are harvested and incorporation of 3H-thymidine used as a measure of proliferation. Anti-CD3 alone is the positive control for proliferation. IL-2 (100 U/ml) is also used as a control which enhances proliferation.
Control antibody which does not induce proliferation of T cells is used as the negative controls for the effects of TNF Delta and/or TNF Epsilon proteins.
Alternatively, a proliferation assay on resting PBL (peripheral blood lymphocytes) is measured by the up-take of 3H-thymidine. The assay is performed as follows. PBMC are isolated by Ficoll (LSM, ICN Biotechnologies, Aurora, Ohio) gradient centrifugation from human peripheral blood, and are cultured overnight in 10% (Fetal Calf Serum, Biofluids, Rockville, MD)1RPMI (Gibco BRL, Gaithersburg, MD). This overnight incubation period allows the adherent cells to attach to the plastic, which results in a lower background in the assay as there are fewer cells that can act as antigen presenting cells or that might be producing growth factors.
The following day the non-adherent cells are collected, washed and used in the proliferation assay. The assay is performed in a 96 well plate using 2 x 104 cells/well in a final volume of 200 microliters. The supernatants (e.g., CHO or 293T
supernatants) expressing the protein of interest are tested at a 30% final dilution, therefore 60u1 are added to 140u1 of 10% FCS/RPMI containing the cells.
Control supernatants are used at the same final dilution and express the following proteins:
vector (negative control), IL-2 (*), IFN , TNF , IL-10 and TR2. In addition to the control supernatants, recombinant human IL-2 (R & D Systems, Minneapolois, MN) at a final concentration of 100ng/ml is also used. After 24 hours of culture, each well is pulsed with luCi of 3H-thymidine (Nen, Boston, MA). Cells are then harvested 20 hours following pulsing and incorporation of 3H-thymidine is used as a measure of proliferation. Results are expressed as an average of triplicate samples plus or minus standard error.
(*) The amount of the control cytokines IL-2, IFN , TNF and IL-10 produced in each transfection varies between 300pg to 5ng/ml.
.
Costimulation assay.

A costimulation assay on resting PBL (peripheral blood lymphocytes) is performed in the presence of immobilized antibodies to CD3 and CD28. The use of antibodies specific for the invariant regions of CD3 mimic the induction of T
cell activation that would occur through stimulation of the T cell receptor by an antigen.
Cross-linking of the TCR (first signal) in the absence of a costimulatory signal (second signal) causes very low induction of proliferation and will eventually result in a state of "anergy", which is characterized by the absence of growth and inability to produce cytokines. The addition of a costimulatory signal such as an antibody to CD28, which mimics the action of the costimulatory molecule. B7-1 expressed on activated APCs, results in enhancement of T cell responses including cell survival and production of IL-2. Therefore this type of assay allows to detect both positive and negative effects caused by addition of supernatants expressing the proteins of interest on T cell proliferation.
The assay is performed as follows. Ninety-six yell plates are coated with ~ 100ng/ml anti-CD3 and Sug/ml anti-CD28 (Pharmingen, San Diego, CA) in a final volume of 100u1 and incubated overnight at 4C. Plates are washed twice with PBS
before use. PBMC are isolated by Ficoll (LSM, ICN Biotechnologies, Aurora, Ohio) gradient centrifugation from human peripheral blood, and are cultured overnight in 10%-FCS(Fetal Calf Serum, Biofluids, Rockville, MD)/RPMI (Gibco BRL, Gaithersburg, MD). This overnight incubation period allows the adherent cells to attach to the plastic, which results in a lower background in the assay as there are fewer cells that can act as antigen presenting cells or that might be producing growth factors. The following day the non adherent cells are collected, washed and used in the proliferation assay. The assay is performed in a 96 well plate using 2 x cells/well in a final volume of 200u1. The supernatants (e.g., CHO or 293T
supernatants) expressing the protein of interest are tested at a 30% final dilution, therefore 60u1 are added to 140u1 of 10% FCS/RPMI containing the cells.
Control supernatants are used at the same final dilution and express the following proteins:
vector only (negative control), IL-2, IF'N , TNF , IL-10 and TR2. In addition to the control supernatants recombinant human IL-2 (R & D Systems, Minneapolis, MN) at a final concentration of l Ong/ml is also used. After 24 hours of culture, each well is pulsed with luCi of 3H-thymidine (Nen, Boston, MA). Cells are then harvested 20 _. .

hours following pulsing and incorporation of 3H-thymidine is used as a measure of proliferation. Results are expressed as an average of triplicate samples plus or minus standard error.
Costimulation assay: IFN y and IL-2 ELISA
The assay is performed as follows. Twenty-four well plates are coated with either 300ng/ml or 600ng/ml anti-CD3 and Sug/ml anti-CD28 (Pharmingen, San Diego, CA) in a final volume of SOOuI and incubated overnight at 4C. Plates are washed twice with PBS before use. PBMC are isolated by Ficoll (LSM, ICN
Biotechnologies, Aurora, Ohio) gradient centrifugation from human peripheral blood, and are cultured overnight in 10% FCS(Fetal Calf Serum, Biofluids, Rockville, MD)/RPMI (Gibco BRL, Gaithersburg, MD). This overnight incubation period allows the adherent cells to attach to the plastic, which results in a lower background in the assay as there are fewer cells that can act as antigen presenting cells or that might be producing growth factors. The following day the non adherent cells are collected, washed and used in the costimulation assay. The assay is performed in the pre-coated twenty-four well plate using 1 x 105 cells/well in a final volume of 900u1.
The supernatants (293T supernatants) expressing the protein of interest are tested at a 30% final dilution, therefore 300u1 are added to 600u1 of 10% FCS/RPMI
containing the cells. Control supernatants are used at the same final dilution and express the following proteins: vector only(negative control), IL-2, IFN , IL-12 and IL-18. In addition to the control supernatants recombinant human IL-2 (all cytokines were purchased from R & D Systems, Minneapolis, MN) at a final concentration of lOng/ml, IL-12 at a final concentration of lng/ml and IL-18 at a final concentration of SOng/ml are also used. Controls and unknown samples are tested in duplicate.
Supernatant samples (250u1) are collected 2 days and 5 days after the beginning of the assay. ELISAs to test for IFN and IL-2 secretion are performed using kits purchased from R & D Systems, (Minneapolis, MN). Results are expressed as an average of duplicate samples plus or minus standard error.
Proliferation assay for preactivated-resting T cells.

A proliferation assay on preactivated-resting T cells is performed on cells that are previously activated with the lectin phytohemagglutinin (PHA). Lectins are polymeric plant proteins that can bind to residues on T cell surface glycoproteins including the TCR and act as polyclonal activators. PBLs treated with PHA and then cultured in the presence of low doses of IL-2 resemble effector T cells. These cells are generally more sensitive to further activation induced by growth factors such as IL-2. This is due to the expression of high affinity IL-2 receptors that allows this population to respond to amounts of IL-2 that are 100 fold lower than what would have an effect on a-naive T cell. Therefore the use of this type of cells might enable to detect the effect of very low doses of an unknown growth factor, that would not be sufficient to induce proliferation on resting (naive ) T cells.
The assay is performed as follows. PBMC are isolated by F/H gradient centrifugation from human peripheral blood, and are cultured inl0% FCS(Fetal Calf Serum, Biofluids, Rockville, MD)/RPMI (Gibco BRL, Gaithersburg, MD) in the presence of 2ug/ml PHA (Sigma, Saint Louis, MO) for three days. The cells are then washed in PBS and cultured inl0% FCS/RPMI in the presence of Sng/ml of human recombinant IL-2 (R & D Systems, Minneapolis, MN) for 3 days. The cells are washed and rested in starvation medium (1%FCS/RPMI) forl6 hours prior to the beginning of the proliferation assay. An aliquot of the cells is analyzed by FACS to determine the percentage of T cells (CD3 positive cells) present; this usually ranges between 93-97% depending on the donor. The assay is performed in a 96 well plate using 2 x104 cells/well in a final volume of 200u1. The supernatants (e.g., CHO or 293T supernatants) expressing the protein of interest are tested at a 30%
final dilution, therefore 60u1 are added to 140u1 of inl0% FCS/RPMI containing the cells.
Control supernatants are used at the same final dilution and express the following proteins:
vector (negative control), IL-2, IFN , TNF , IL-10 and TR2. In addition to the control supernatants recombinant human IL-2 at a final concentration of lOng/ml is also used. After 24 hours of culture, each well is pulsed with luCi of 3H-thymidine(Nen, Boston, MA). Cells are then harvested 20 hours following pulsing and incorporation of 3H-thymidine is used as a measure of proliferation.
Results are expressed as an average of triplicate samples plus or minus standard error.

The studies described in this example test activity of polypeptides of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides of the invention (e.g., gene therapy), agonists, and/or antagonists of polynucleotides or polypeptides of the invention.
Example 34: Effect of Polypeptides of the Invention on the Expression of MHC
Class II, Costimulatory and Adhesion Molecules and Cell Differentiation of Monocytes and Monocyte-Derived Human Dendritic Cells Dendritic cells are generated by the expansion of proliferating precursors found in the peripheral blood: adherent PBMC or elutriated monocytic fractions are cultured for 7-10 days with GM-CSF (50 ng/ml) and IL-4 (20 ng/ml). These dendritic cells have the characteristic phenotype of immature cells (expression of CD1, CD80, CD86, CD40 and MHC class II antigens). Treatment with activating factors, such as TNF-a, causes a rapid change in surface phenotype (increased expression of MHC class I and II, costimulatory and adhesion molecules, downregulation of FCyRII, upregulation of CD83). These changes correlate with increased antigen-presenting capacity and with functional maturation of the dendritic cells.
FACS analysis of surface antigens is performed as follows. Cells are treated 1-days with increasing concentrations of polypeptides of the invention or LPS
(positive control), washed with PBS containing 1 % BSA and 0.02 mM sodium azide, and then incubated with 1:20 dilution of appropriate FITC- or PE-labeled monoclonal antibodies for 30 minutes at 4 degrees C. After an additional wash, the labeled cells are analyzed by flow cytometry on a FACScan (Becton Dickinson).
Effect on the production of cvtokines. Cytokines generated by dendritic cells, in particular IL-12, are important in the initiation of T-cell dependent immune responses. IL-12 strongly influences the development of Thl helper T-cell immune response, and induces cytotoxic T and NK cell function. An ELISA is used to measure the IL-12 release as follows. Dendritic cells (106/m1) are treated with increasing concentrations of polypeptides of the invention for 24 hours. LPS
(100 ng/ml) is added to the cell culture as positive control. Supernatants from the cell cultures are then collected and analyzed for IL-12 content using commercial ELISA
kit (e..g, R & D Systems (Minneapolis, MN)). The standard protocols provided with the kits are used.
Effect on the expression of MHC Class II, costimulatory and adhesion molecules. Three major families of cell surface antigens can be identified on monocytes: adhesion molecules, molecules involved in antigen presentation, and Fc receptor. Modulation of the expression of MHC class II antigens and other costimulatory molecules, such as B7 and ICAM-1, may result in changes in the antigen presenting capacity of monocytes and ability to induce T cell activation.
Increase expression of Fc receptors may correlate with improved monocyte cytotoxic activity, cytokine release and phagocytosis.
FACS analysis is used to examine the surface antigens as follows. Monocytes are treated 1-5 days with increasing concentrations of polypeptides of the invention or LPS (positive control), washed with PBS containing 1% BSA and 0.02 mM sodium azide, and then incubated with 1:20 dilution of appropriate FITC- or PE-labeled monoclonal antibodies for 30 minutes at 4 degreesC. After an additional wash, the labeled cells are analyzed by flow cytometry on a FACScan (Becton Dickinson).
Monocyte activation and/or increased survival. Assays for molecules that activate (or alternatively, inactivate) monocytes and/or increase monocyte survival (or alternatively, decrease monocyte survival) are known in the art and may routinely be applied to determine whether a molecule of the invention functions as an inhibitor or activator of monocytes. Polypeptides, agonists, or antagonists of the invention can be screened using the three assays described below: For each of these assays, Peripheral blood mononuclear cells (PBMC) are purified from single donor leukopacks (American Red Cross, Baltimore, MD) by centrifugation through a Histopaque gradient (Sigma): Monocytes are isolated from PBMC by counterflow centrifugal elutriation.
Monocyte Survival Assay. Human peripheral blood monocytes progressively lose viability when cultured in absence of serum or other stimuli. Their death results from internally regulated process (apoptosis). Addition to the culture of activating factors, such as TNF-alpha dramatically improves cell survival and prevents DNA
fragmentation. Propidium iodide (PI) staining is used to measure apoptosis as follows. Monocytes are cultured for 48 hours in polypropylene tubes in serum-free medium (positive control), in the presence of 100 ng/ml TNF-alpha (negative control), and in the presence of varying concentrations of the compound to be tested.
Cells are suspended at a concentration of 2 x 106/m1 in PBS containing PI at a final concentration of 5 p.g/ml, and then incubaed at room temperature for 5 minutes before FACScan analysis. PI uptake has been demonstrated to correlate with DNA
fragmentation in this experimental paradigm.
Effect on cytokine release. An important function of monocytes/macrophages is their regulatory activity on other cellular populations of the immune system through the release of cytokines after stimulation. An ELISA to measure cytokine release is performed as follows. Human monocytes are incubated at a density of Sx105 cells/ml 1 S with increasing concentrations of the a polypeptide of the invention and under the same conditions, but in the absence of the polypeptide. For IL-12 production, the cells are primed overnight with IFN (100 U/ml) in presence of a polypeptide of the invention. LPS (10 ng/ml) is then added. Conditioned media are collected after 24h and kept frozen until use. Measurement of TNF-alpha, IL-10, MCP-1 and IL-8 is then performed using a commercially available ELISA kit (e..g, R & D Systems (Minneapolis, MN)) and applying the standard protocols provided with the kit.
Oxidative burst. Purified monocytes are plated in 96-w plate at 2-1x105 cell/well. Increasing concentrations of polypeptides of the invention are added to the wells in a total volume of 0.2 ml culture medium (RPMI 1640 + 10% FCS, glutamine and antibiotics). After 3 days incubation, the plates are centrifuged and the medium is removed from the wells. To the macrophage monolayers, 0.2 ml per well of phenol red solution (140 mM NaCI, 10 mM potassium phosphate buffer pH 7.0, 5.5 mM
dextrose, 0.56 mM phenol red and 19 U/ml of HRPO) is added, together with the stimulant (200 nM PMA). The plates are incubated at 37°C for 2 hours and the reaction is stopped by adding 20 p.1 1 N NaOH per well. The absorbance is read at 610 nm. To calculate the amount of H202 produced by the macrophages, a standard curve of a H20z solution of known molarity is performed for each experiment.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polypeptides, polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 35: Biological Effects of Polypeptides of the Invention ' Astrocyte and Neuronal Assays Recombinant polypeptides of the invention; expressed in Escherichia coli and purified as described above, can be tested for activity in promoting the survival, neurite outgrowth, or phenotypic differentiation of cortical neuronal cells and for inducing the proliferation of glial fibrillary acidic protein immunopositive cells, astrocytes. The selection of cortical cells for the bioassay is based.on the prevalent expression of FGF-1 ~d FGF-2 in cortical structures and on the previously reported enhancement of cortical neuronal survival resulting from FGF-2 treatment. A thymidine incorporation assay, for example, can be used to elucidate a polypeptide of the invention's activity on these cells.
Moreover, previous reports describing the biological effects of FGF-2 (basic FGF) on cortical or hippocampal neurons in vitro have demonstrated increases in both neuron survival and neurite outgrowth (Walicke et al., "Fibroblast growth factor promotes survival of dissociated hippocampal neurons and enhances neurite extension."
Proc. Natl.
Acad. Sci. USA 83:3012-3016. (1986), assay herein incorporated by reference in its entirety). However, reports from experiments done on PC-12 cells suggest that these two responses are not necessarily synonymous and may depend on not only which FGF
is being tested but also on which receptors) are expressed on the target cells.
Using the primary cortical neuronal culture paradigm, the ability of a polypeptide of the invention to induce neurite outgrowth can be compared to the response achieved with FGF-2 using, for example, a thymidine incorporation assay.
Fibroblast and endothelial cell assays~

Human lung fibroblasts are obtained from Clonetics (San Diego, CA) and maintained in growth media from Clonetics. Dermal microvascular endothelial cells are obtained from Cell Applications (San Diego, CA). For proliferation assays, the human lung fibroblasts and dermal microvascular endothelial cells can be cultured at 5,000 cells/well in a 96-well plate for one day in growth medium. The cells are then incubated for one day in 0.1% BSA basal medium. After replacing the medium with fresh 0.1% BSA
medium, the cells are incubated with the test proteins for 3 days. Alamar Blue (Alamar Biosciences, Sacramento, CA) is added to each well to a final concentration of 10%. The cells are incubated for 4 hr. Cell viability is measured by reading in a CytoFluor fluorescence reader. For the PGEz assays, the human lung fibroblasts are cultured at 5,000 cells/well in a 96-well plate for one day. After a medium change to 0.1 % BSA
basal medium, the cells are incubated with FGF-2 or polypeptides of the invention with or without IL-la for 24 hours. The supernatants are collected and assayed for PGEZ by EIA
kit (Cayman, Ann Arbor, MI). For the IL-6 assays, the human lung fibroblasts are cultured at 5,000 cells%vvell in a 96-well plate for one day. After a medium change to 0.1% BSA basal medium, the cells are incubated with FGF-2 or with or without polypeptides of the invention IL-la for 24 hours. The supernatants are collected and assayed for IL-6 by ELISA kit (Endogen, Cambridge, MA).
Human lung fibroblasts are cultured with FGF-2 or polypeptides of the invention for 3 days in basal medium before the addition of Alamar Blue to assess effects on growth of the fibroblasts. FGF-2 should show a stimulation at 10 - 2500 ng/ml which can be used to compare stimulation with polypeptides of the invention.
Parkinson Models.
The loss of motor function in Parkinson's disease is attributed to a deficiency of striatal dopamine resulting from the degeneration of the nigrostriatal dopaminergic projection neurons. An animal model for Parkinson's that has been extensively characterized involves the systemic administration of 1-methyl-4 phenyl 1,2,3,6-tetrahydropyridine (MPTP). In the CNS, MPTP is taken-up by astrocytes and catabolized by monoamine oxidase B to 1-methyl-4-..phenyl pyridine (MPP+) and released.

Subsequently, MPP+ is actively accumulated in dopaminergic neurons by the high-affinity reuptake transporter for dopamine. MPP+ is then concentrated in mitochondria by the electrochemical gradient and selectively inhibits nicotidamide adenine disphosphate:
ubiquinone oxidoreductionase (complex I), thereby interfering with electron transport and eventually generating oxygen radicals.
It has been demonstrated in tissue culture paradigms that FGF-2 (basic FGF) has trophic activity towards nigral dopaminergic neurons (Ferrari et al., Dev.
Biol. 1989).
Recently, Dr. Unsicker's group has demonstrated that administering FGF-2 in gel foam implants in the striatum results in the near complete protection of nigral dopaminergic neurons from the toxicity associated with MPTP exposure (Otto and Unsicker, J.
Neuroscience, 1990).
Based on the data with FGF-2, polypeptides of the invention can be evaluated to determine whether it has an action similar to that of FGF-2 in enhancing dopaminergic neuronal survival in vitro and it can also be tested in vivo for protection of dopaminergic neurons in the striatum from the damage associated with MPTP treatment. The potential effect of a polypeptide of the invention is first examined in vitro in a dopaminergic neuronal cell culture paradigm. The cultures are prepared by dissecting the midbrain floor plate from gestation day 14 Wistar rat embryos. The tissue is dissociated with trypsin and seeded at a density of 200,000 cells/cm2 on polyorthinine-laminin coated glass coverslips.
The cells are maintained in Dulbecco's Modified Eagle's medium and F12 medium containing hormonal supplements (N1). The cultures are fixed with paraformaldehyde after 8 days in vitro and are processed for tyrosine hydroxylase, a specific marker for dopminergic neurons, immunohistochemical staining. Dissociated cell cultures are prepared from embryonic rats. The culture medium is changed every third day and the factors are also added at that time.
Since the dopaminergic neurons are isolated from animals at gestation day 14, a developmental time which is past the stage when the dopaminergic precursor-cells are proliferating, an increase in the number of tyrosine hydroxylase immunopositive neurons would represent an increase in the number of dopaminergic neurons surviving in vitro.
Therefore, if a polypeptide of the invention acts to prolong the survival of dopaminergic neurons, it would suggest that- the polypeptide may be involved in Parkinson's Disease.

The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 36: The Effect of Polypeptides of the Invention on the Growth of Vascular Endothelial Cells On day 1, human umbilical vein endothelial cells (HUVEC) are seeded at 2-5x104 cells/35 mm dish density in M199 medium containing 4% fetal bovine serum (FBS), 16 units/ml heparin, and 50 units/ml endothelial cell growth supplements (ECGS, Biotechnique, Inc.). On day 2, the medium is replaced with M199 containing 10%
FBS, 8 units/ml heparin. A polypeptide having the amino acid sequence of SEQ ff~
NO:Y, and positive controls, such as VEGF and basic FGF (bFGF) are added, at varying . concentrations. On days 4 and 6, the medium is replaced. On day 8, cell number is determined with a Coulter Counter.
An increase in the number of HUVEC cells indicates that the polypeptide of the invention may proliferate vascular endothelial cells.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 37: Stimulatory Effect of Polypeptides of the Invention on the Proliferation of Vascular Endothelial Cells For evaluation of mitogenic activity of growth factors, the colorimetric MTS
(3-(4,5-dimethylthiazol-2-yl)-S-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium) assay with the electron coupling reagent PMS (phenazine methosulfate) was performed (CellTiter 96 AQ; Promega). Cells are seeded in a 96-well plate (5,000 cells/well) in 0.1 mL serum-supplemented medium and are allowed to attach overnight.
After serum-starvation for 12 hours in 0.5% FBS, conditions (bFGF, VEGFibs or a polypeptide of the invention in 0.5% FBS) with or without Heparin (8 U/ml) are added to wells for 48 hours. 20 mg of MTS/PMS mixture (1:0.05) are added per well and allowed to incubate for 1 hour at 37°C before measuring the absorbance at 490 nm in an ELISA
plate reader. Background.absorbance from control wells (some media, no cells) is subtracted, and seven wells are performed in parallel for each condition. See, Leak et al.
In Vitro Cell. Dev. Biol. 30A:512-518 (1994).
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the .
invention.
Example 38: Inhibition of PDGF-induced Vascular Smooth Muscle Cell Proliferation Stimulatory Effect HAoSMC proliferation can be measured, for example, by BrdUrd incorporation.
Briefly, subconfluent, quiescent cells grown_on the 4-chamber slides are transfected with CRP or FITC-labeled AT2-3LP. Then, the cells are pulsed with 10% calf serum and 6 mg/ml BrdUrd. After 24 h, immunocytochemistry is performed by using BrdUrd Staining Kit (Zymed Laboratories). In brief, the cells are incubated with the biotinylated mouse anti-BrdUrd antibody at 4 degrees C for 2 h after being exposed to denaturing solution and then incubated with the streptavidin-peroxidase and diaminobenzidine. After counterstaining with hematoxylin, the cells are mounted for microscopic examination, and the BrdUrd-positive cells are counted. The BrdUrd index is calculated as a percent of the BrdUrd-positive cells to the total cell number. In addition, the simultaneous detection of the BrdUrd staining (nucleus) and the FITC uptake (cytoplasm) is performed for individual cells by the concomitant use of bright field illumination and dark field-UV
fluorescent illumination. See, Hayashida et al., J. Biol. Chem.
6:271(36):21985-21992 ( 1996).
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.

Example 39: Stimulation of Endothelial Migration This example will be used to explore the possibility that a polypeptide of the invention may stimulate lymphatic endothelial cell migration.
Endothelial cell migration assays are performed using a 48 well microchemotaxis chamber (Neuroprobe Inc:, Cabin John, MD; Falk, W., et al., J. Immunological Methods 1980;33:239-247). Polyvinylpyrrolidone-free polycarbonate filters with a pore size of 8 um (Nucleopore Corp. Cambridge, MA) are coated with 0.1% gelatin for at least 6 hours at room temperature and dried under sterile air. Test substances are diluted to appropriate concentrations in M199 supplemented with 0.25% bovine serum albumin (BSA), and ~ul of the final dilution is placed in the lower chamber of the modified Boyden apparatus.
Subconfluent, early passage (2-6) HWEC or BMEC cultures are washed and trypsinized for the minimum time required to achieve cell detachment. After placing the filter between lower and upper chamber, 2.5 x 105 cells suspended in 50 u1 M199 containing 1%
FBS are seeded in the upper compartment. The apparatus is then incubated for.
S hours at 37°C in a humidified chamber with 5% C02 to allow cell migration. After the incubation period, the filter is removed and the upper side of the filter with the non-migrated cells is scraped with a rubber policeman. The filters are fixed with methanol and stained with a Giemsa solution (Diff Quick, Baxter, McGraw Park, IL). Migration is quantified by counting cells of three random high-power fields (40x) in each well, and all groups are performed in quadruplicate.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 40: Stimulation of Nitric Oxide Production by Endothelial Cells Nitric oxide released,by the vascular endothelium is believed to be a mediator of vascular endothelium relaxation. Thus, activity of a polypeptide of the invention can be assayed by determining nitric oxide production by endothelial cells in response to the polypeptide.
Nitric oxide is measured in 96-well plates of confluent microvascular endothelial cells after 24 hours starvation and a subsequent 4 hr exposure to various levels of a positive control (such as VEGF-1) and the polypeptide of the invention. Nitric oxide in the medium is determined by use of the Griess reagent to measure total nitrite after reduction of nitric oxide-derived nitrate by nitrate reductase. The effect of the polypeptide of the invention on nitric oxide release is examined on HUVEC.
J
Briefly, NO release from cultured HLTVEC monolayer is measured with a NO-specific polarographic electrode connected to a NO meter (Iso-NO, World Precision Instruments Inc.) (1049). Calibration of the NO elements is performed according to the following equation:
2KN0z+2KI+2HZS0462N0+Iz+2Hz0+2KZS04 The standard calibration curve is obtained by adding graded concentrations of KN02 (0, 5, 10, 25, 50, 100, 250, and 500 nmol/L) into the calibration solution containing KI and HZS04. The specificity of the Iso-NO electrode to NO is previously 'determined by measurement of NO from authentic NO gas (1050). The culture medium is removed and HUVECs are washed twice with Dulbecco's phosphate buffered saline. The cells are then bathed in S ml of filtered Krebs-Henseleit solution in 6-well plates, and the cell plates are kept on a slide warmer (Lab Line Instruments Inc.) To maintain the temperature at 37°C.
The NO sensor probe is inserted vertically into the wells, keeping the tip of the electrode 2 mm under the surface of the solution, before addition of the different conditions.
S-nitroso acetyl penicillamin (SNAP) is used as a positive control. The amount of released NO is expressed as picomoles per 1x106 endothelial cells: All values reported are means of four to six measurements in each group (number of cell culture wells). See, Leak et al. Biochem. and Biophys. Res..Comm. 217:96-105 (1995).
The studies described in this example tested activity of polypeptides of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.

Example 41: Effect of Polypepides of the Invention on Cord Formation in An~io~enesis Another step in angiogenesis is cord formation, marked by differentiation of endothelial cells. This bioassay measures the ability of microvascular endothelial cells to form capillary-like structures (hollow structures) when cultured in vitro.
CADMEC (microvascular endothelial cells) are purchased from Cell Applications, Inc. as proliferating (passage 2) cells and are cultured in Cell Applications' CADMEC
Growth Medium and used at passage S. For the in vitro angiogenesis assay, the wells of a 48-well cell culture plate are coated with Cell Applications' Attachment Factor Medium (200 ml/well) for 30 min. at 37°C. CADMEC are seeded onto the coated wells at 7,500 cells/well and cultured overnight in Growth Medium. The Growth Medium is then replaced with 300 mg Cell Applications' Chord Formation Medium containing control buffer or a polypeptide of the invention (0.1 to 100 ng/ml) and the cells are cultured for an additional 48 hr. The numbers and lengths of the capillary-like.chords are quantitated through use of the Boeckeler VIA-170 video image analyzer. All assays are done in triplicate.
Commercial (R&D) VEGF (50 nglml) is used as a positive control. b-esteradiol (1 ng/ml) is used as a negative control. The appropriate buffer (without protein) is also utilized as a control.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 42: An~io~enic Effect on Chick Chorioallantoic Membrane Chick chorioallantoic membrane (CAM) is a well-established system to examine angiogenesis. Blood vessel formation on CAM is easily visible and quantifiable. The ability of polypeptides of the invention to stimulate angiogenesis in CAM can be examined.

Fertilized eggs of the White Leghorn chick (callus gallus) and the Japanese qual (Coturnix coturnix) are incubated at 37.8°C and 80% humidity.
Differentiated CAM of 16-day-old chick and 13-day-old qual embryos is studied with the following methods.
On Day 4 of development, a window is made into the egg shell of chick eggs.
The embryos are checked for normal development and the eggs sealed with cellotape.
They are further incubated until Day 13. Thermanox coverslips (Nunc, Naperville, IL) are cut into disks of about 5 mm in diameter. Sterile and salt-free growth factors are dissolved iri distilled water and about 3.3 mg/ 5 ml are.pipetted on the disks. After air-drying, the inverted disks are applied on CAM. After 3 days, the specimens are fixed in 3%
glutaraldehyde and 2% formaldehyde and rinsed in 0.12 M sodium cacodylate buffer.
They are photographed with a stereo microscope [Wild M8] and embedded for semi-and ultrathin sectioning as described above. Controls are performed with carrier disks alone.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to 1 S test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 43: An~io~enesis Assay Using a Matri~el Implant in Mouse In vivo angiogenesis assay of a polypeptide of the invention measures the ability of an existing capillary network to form new vessels in an implanted capsule of murine extracellular matrix material (Matrigel). The protein is mixed with the liquid Matrigel at 4 degree C and the mixture is then injected subcutaneously in mice where it solidifies. After 7 days, the solid "plug" of Matrigel is removed and examined for the presence of new blood vessels. Matrigel is purchased from Becton Dickinson Labware/Collaborative Biomedical Products.
When thawed at 4 degree C the Matrigel material is a liquid. The Matrigel is mixed with a polypeptide of the invention at 150 ng/ml at 4 degrees C and drawn into cold 3 ml syringes. Female C57B1/6 mice approximately 8 weeks old are injected with the mixture of Matrigel and experimental protein at 2 sites at the midventral aspect of the abdomen (0.5 ml/site). A$er 7 days, the mice are sacrificed by cervical dislocation, the Matrigel plugs are removed and cleaned (i.e., all clinging membranes and fibrous tissue is removed). Replicate whole plugs are fixed in neutral buffered 10%
formaldehyde, embedded in paraffin and used to produce sections for histological examination after staining with Masson's Trichrome. Cross sections from 3 different regions of each plug are processed. Selected sections are stained for the presence of v,WF. The positive control for this assay is bovine basic FGF (150 ng/ml). Matrigel alone is used to determine basal levels of angiogenesis.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 44: Rescue of Ischemia in Rabbit Lower Limb Model To study the in vivo effects of polynucleotides and polypeptides of the invention on ischemia, a rabbit hindlimb ischemia model is created by surgical removal of one femoral arteries as described previously (Takeshita et al., Am J. Pathol 147:1649-1660 (1995)). The excision of the femoral artery results in retrograde propagation of thrombus and occlusion of the external iliac artery. Consequently, blood flow to the ischemic limb is dependent upon collateral vessels originating from the internal iliac artery (Takeshitaet al. Am J. Pathol 147:1649-1660 (1995)). An interval of 10 days is allowed for post-operative recovery of rabbits and development of endogenous collateral vessels. At 10 day post-operatively (day 0), after performing a baseline angiogram, the internal i 1 iac artery of the ischemic limb is transfected with 500 mg naked expression plasmid containing a polynucleotide of the invention by arterial gene transfer technology using a hydrogel-coated balloon catheter as described (Riessen et al. Hum Gene Ther.
4:749-758 (1993); Leclerc et al. J. Clin. Invest. 90: 936-944 (1992)). When a polypeptide of the invention is used in the treatment, a single bolus of 500 mg polypeptide of the invention or control is delivered into the internal iliac artery of the ischemic limb over a period of 1 min. through an infusion catheter. On day 30, various parameters are measured in these rabbits: (a) BP ratio - The blood pressure ratio of systolic pressure of the ischemic limb to that of normal limb; (b) Blood Flow. and Flow Reserve - Resting FL: the blood flow during undilated condition and Max FL: the blood flow during fully dilated condition (also an indirect measure of the blood vessel amount) and Flow Reserve is reflected by the ratio of max FL: resting FL; (c) Angiographic Score - This is measured by the angiogram of collateral vessels. A score is determined by the percentage of circles in an overlaying grid that with crossing opacified arteries divided by the total number m the rabbit thigh; (d) Capillary density - The number of collateral capillaries determined in light microscopic .
sections taken from hindlimbs.
The studies described in this example tested activity of polynucleotides and polypeptides of the invention. However, one skilled in the art could' easily modify the exemplified studies to test the agonists, and/or antagonists of the invention.
Example 45: Effect of Polypeptides of the Invention on Vasodilation Since dilation of vascular endothelium is important in reducing blood pressure, the ability of polypeptides of the invention to affect the blood pressure in spontaneously hypertensive rats (SHR) is examined. Increasing doses (0, 10, 30, 100, 300, and 900 mg/kg) of the polypeptides of the invention are administered to 13-14 week old spontaneously hypertensive rats (SHR). Data are expressed as the mean +/- SEM.
Statistical analysis are performed with a paired t-test and statistical significance is defined as p<0.05 vs. the response to buffer alone.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modifythe exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 46: Rat Ischemic Skin Flap Model The evaluation parameters include skin blood flow, skin temperature, and factor VIII immunohistochemistry or endothelial alkaline phosphatase reaction.
Expression of polypeptides of the invention, during the skin ischemia, is studied using in situ hybridization.
The study in this model is divided into three parts as follows:

Ischemic skin Ischemic skin wounds Normal wounds The experimental protocol includes:
Raising a 3x4 cm, single pedicle full-thickness random skin flap (myocutaneous flap over the lower back of the animal).
An excisional wounding (4-6 mm in diameter) in the ischemic skin (skin-flap).
Topical treatment with a polypeptide of the invention of the excisional wounds (day 0, 1, 2, 3, 4 post-wounding) at the following various dosage ranges: lmg to 100 mg.
Harvesting the wound tissues at day 3, S, 7, 10, 14 and 21 post-wounding for histological, immunohistochemical, and in situ studies.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 47: Peripheral Arterial Disease Model Angiogenic therapy using a polypeptide of the invention is a novel therapeutic strategy to obtain restoration of blood flow around the ischemia in case of peripheral arterial diseases. The experimentaj protocol includes:
One side of the femoral artery is ligated to create ischemic muscle of the hindlimb, the other side of hindlimb serves as a control. _ a polypeptide of the invention, in a dosage range of 20 mg - 500 mg, is delivered intravenously and/or intramuscularly 3 times (perhaps more) per week for 2-3 weeks.
The ischemic muscle tissue is collected after ligation of the femoral artery at 1, 2, and 3 weeks for the analysis of expression of a polypeptide of the invention and histology. Biopsy is also performed on the other side of normal muscle of the contralateral hindlimb.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 48: Ischemic Myocardial Disease Model A polypeptide of the invention is evaluated as a potent mitogen capable of stimulating the development of collateral vessels, and restructuring new vessels after coronary artery occlusion. Alteration of expression of the polypeptide is investigated in situ. The experimental protocol includes:
The heart is exposed through a left-side thoracotomy in the rat. Immediately, the left coronary artery is occluded with a thin suture (6-0) and the thorax is closed.
a polypeptide of the invention, in a dosage range of 20 mg - 500 mg, is delivered intravenously and/or intramuscularly 3 times (perhaps more) per week for 2-4 weeks.
Thirty days after the surgery, the heart is removed and cross-sectioned for morphometric and in situ analyzes.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 49: Rat Corneal. Wound Healing Model This animal model shows the effect of a polypeptide of the invention on neovascularization. The eXperimental protocol includes:
Making a 1-1.5 mm long incision from the center of cornea into the stromal layer.
Inserting a spatula below the lip of the incision facing the outer corner of the eye.
Making a pocket (its base is 1-1.5 mm form the edge of the eye).
Positioning a pellet, containing SOng- Sug of a polypeptide of the invention, within the pocket.
Treatment with a polypeptide of the invention can also be applied topically to the corneal wounds in a dosage range of 20mg - SOOmg (daily treatment for five days).

The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 50: Diabetic Mouse and Glucocorticoid-Impaired Wound Healing Models Diabetic db+/db+ Mouse Model.
To demonstrate that a polypeptide of the invention accelerates the healing process, the genetically diabetic mouse model of wound healing is used. The full thickness wound healing model in the db+/db+ mouse is a well characterized, clinically relevant and reproducible model of impaired wound healing: Healing of the diabetic wound is dependent on formation of granulation tissue and re-epithelialization rather than contraction (Gartner, M.H. et al., J. Surg. Res. 52:389 (1992); Greenhalgh, D.G. et al., Am. J. Pathol. 136:1235 (1990)).
The diabetic animals have many of the characteristic features observed in Type II
diabetes mellitus. Homozygous (db+/db+) mice are obese in comparison to their normal heterozygous (db+/+m) littermates. Mutant diabetic (db+/db+) mice have a single autosomal recessive mutation on chromosome 4 (db+) (Coleman et al. Proc. Natl.
Acad.
Sci. USA 77:283-293 (1982)). Animals show polyphagia, polydipsia and polyuria.
Mutant diabetic mice (db+/db+) have elevated blood glucose, increased or normal insulin levels, and suppressed cell-mediated immunity (Mandel et al., J. Immunol.
120:1375 (1978); Debray-Sachs, M. et al., Clin. Exp. Immunol. 51 (1):1-7 (1983); Leiter et al., Am.
J. ofPathol. 114:46-55 (1985)). Peripheral neuropathy, myocardial complications, and microvascular lesions, basement membrane thickening and glomerular filtration abnormalities have been described in these animals (Norido, F. et al., Exp.
Neurol.
83(2):221-232 (1984); Robertson et al., Diabetes 29(1):60-67 (1980);
Giacomelli et al., Lab Invest.~40(4):460-473 (1979); Coleman, D.L., Diabetes 31 (Suppl):1-6 (1982)). These homozygous diabetic mice develop hyperglycemia that is resistant to insulin analogous to human type II diabetes (Mandel et al., J. Immunol. 120:1375-1377 (1978)).

The characteristics observed in these animals suggests that healing in this model may be similar to the healing observed in human diabetes (Greenhalgh, et al., Am. J. of Pathol. 136:1235-1246 (1990)).
Genetically diabetic female C57BL/KsJ (db+/db+) mice and their non-diabetic (db+/+m) heterozygous littermates are used iri this study (Jackson Laboratories). The animals are purchased at 6 weeks of age and are 8 weeks old at the beginning of the study.
Animals are individually housed and received food and water ad libitum. All manipulations are performed using aseptic techniques. The experiments are conducted according to the rules and guidelines of Human Genome Sciences, Inc.
Institutional Animal Care and Use Committee and the Guidelines for the Care and Use of Laboratory Animals.
Wounding protocol is performed according to previously reported methods (Tsuboi, R. and Riflcin, D.B:, J. Exp. Med. 172:245-251 (1990)). Briefly, on the day of wounding, animals are anesthetized with an intraperitoneal injection of Avertin (0.01 mg/mL), 2,2,2-tribromoethanol and 2-methyl-2-butanol dissolved in deionized water. The dorsal region of the animal is shaved and the skin washed with 70% ethanol solution and iodine. The surgical area is dried with sterile gauze prior to wounding. An 8 mm full-thickness wound is then created using a Keyes tissue punch. Immediately following wounding, the surrounding skin is gently stretched to eliminate wound expansion. The ~ wounds are left open for the duration of the experiment. Application of the treatment is given topically for 5 consecutive days commencing on the day of wounding.
Prior to treatment, wounds are gently cleansed with sterile saline and gauze sponges.
' Wounds are visually examined and photographed at a fixed distance at the day of surgery and at two day intervals thereafter. Wound closure is determined by daily measurement on days 1-S and on day 8. Wounds are measured horizontally and vertically using a calibrated Jameson caliper. Wounds are considered healed if granulation tissue is no longer visible and the wound is covered by a continuous epithelium. .
A polypeptide of the invention is administered using at a range different doses, from 4mg to SOOmg per wound per day for 8 days in vehicle. Vehicle control groups received SOmL of vehicle solution.
Animals are euthanized on day 8 with an intraperitoneal injection of sodium pentobarbital (300mg/kg). The wounds and surrounding skin are then harvested for -.

histology and immunohistochemistry. Tissue specimens are placed in 10% neutral buffered formalin in tissue cassettes between biopsy sponges for further processing.
Three groups of 10 animals each (5 diabetic and 5 non-diabetic controls) are evaluated: 1) Vehicle placebo control, 2) untreated group, and 3) treated group.
Wound closure is analyzed by measuring the area in the vertical and horizontal axis and obtaining the total square area of the wound. Contraction is then estimated by establishing the differences between the initial wound area (day 0) and that of post treatment (day 8). The wound area on day 1 is 64mmz, the corresponding size of the dermal punch. Calculations are made using the following formula:
[Open area on day 8] - [Open area on day 1] / [Open area on day 1]
Specimens are fixed in 10% buffered formalin and paraffin embedded blocks are sectioned perpendicular to the wound surface (Smm) and cut using a Reichert-Jung microtome. Routine hematoxylin-eosin (H&E) staining is performed on cross-sections of bisected wounds. Histologic examination of the wounds are used to assess whether the healing process and the morphologic appearance of the repaired skin is altered by treatment with a polypeptide of the invention. This assessment included verification of the presence of cell accumulation, inflammatory cells, capillaries, fbroblasts, re-epithelialization and epidermal maturity (Greenhalgh, D.G. et al., Am. J.
Pathol. 136:1235 ( 1990)). A calibrated lens micrometer is used by a blinded observer.
Tissue sections are also stained immunohistochemically with a polyclonal rabbit anti-human keratin antibody using ABC Elite detection system. Human skin is used as a positive tissue control while non-immune IgG is used as a negative control.
Keratinocyte growth is determined by evaluating the extent of reepithelialization of the wound using a calibrated lens micrometer.
Proliferating cell nuclear antigen/cyclin (PCNA) in skin specimens is demonstrated by using anti-PCNA antibody (1:50) with an ABC Elite detection system. Human colon cancer can serve as a positive tissue control and human brain tissue can.be used as a negative tissue control. Each specimen includes a section with omission of the primary antibody and substitution with non-immune mouse IgG. Ranking of these sections is based on the extent of proliferation on a scale of 0-8, the lower side of the scale reflecting slight proliferation to the higher side reflecting intense proliferation.
Experimental data are analyzed using an unpaired t test. A p value of < 0.05 is considered significant.
Steroid Impaired Rat Model The inhibition of wound healing by steroids has been well documented in various in vitro and in vivo systems (Wahl, Glucocorticoids and Wound healing. In:
Anti-Inflammatory Steroid Action: Basic and Clinical Aspects. 280-302 (1989);
Wahlet al., J.
Immunol. 1l S: 476-481 (1975); Werb et al., J. Exp. Med. 147:1684-1694 (1978)).
Glucocorticoids retard wound healing by inhibiting angiogenesis, decreasing vascular permeability (Ebert et al., An. Intern. Med. 37:701-705 (1952)), fibroblast proliferation-, and collagen synthesis (Beck et al.; Growth Factors. 5: 295-304 (1991); Haynes et al., J. Clin. Invest. 61: 703-797 (1978)) and producing a transient reduction of circulating monocytes (Haynes et al., J. Clin. Invest. 61: 703-797 (1978); Wahl, "Glucocorticoids and wound healing", In: Antiinflammatory Steroid Action: Basic and Clinical Aspects, Academic Press, New York, pp. 280-302 (1989)). The systemic administration of steroids to impaired wound healing is a well establish phenomenon in rats (Beck et al., Growth Factors. 5: 295-304 (1991); Haynes et al., J. Clin. Invest. 61: 703-797 (1978); Wahl, "Glucocorticoids and wound healing", In: Antiinflammatory Steroid Action:
Basic and Clinical Aspects, Academic Press, New York, pp. 280-302 (1989); Pierce et al., Proc.
Natl. Acad. Sci. USA 86: 2229-2233 (1989)).
To demonstrate that a polypeptide of the invention can accelerate the healing process, the effects of multiple topical applications of the polypeptide on full thickness excisional skin wounds in rats in which healing has been impaired by the systemic administration of methylprednisolone is assessed.
Young adult male Sprague Dawley rats weighing 250-300 g (Charles River Laboratories) are used in this example: The animals are purchased at 8 weeks of age and are 9 weeks old at the beginning of the study. The healing response of rats is impaired by the systemic administration of methylprednisolone (l7mg/kg/iat intramuscularly) at the time of wounding. Animals are individually housed and received food and water ad libitum. All manipulations are performed using aseptic techniques. This study is conducted according to the rules and guidelines of Human Genome Sciences, Inc.
Institutional Animal Care and Use Committee and the Guidelines for the Care and Use of Laboratory Animals.
The wounding protocol is followed according to section A, above. On the day of wounding, animals are anesthetized with an intramuscular injection of ketamine (50 mg/kg) and xylazine (5 mg/kg). The dorsal region of the animal is shaved and the skin washed with 70% ethanol and iodine solutions. The surgical area is dried with sterile gauze prior to wounding. An 8 mm full-thickness wound is created using a Keyes tissue punch. The wounds are left open for the duration of the experiment.
Applications ofthe testing~materials are given topically once a day for 7 consecutive days commencing on the day of wounding and subsequent to methylprednisolone administration. Prior to treatment, wounds are gently cleansed with sterile saline and gauze sponges.
Wounds are visually examined and photographed at a fixed distance at the day of wounding and at the end of treatment. Wound closure, is determined by daily measurement on days 1-5 and on day 8. Wounds are measured horizontally and vertically using a calibrated Jameson caliper. Wounds are considered healed if granulation tissue is no longer visible and the wound is covered by a continuous epithelium.
The polypeptide of the invention is administered using at a range different doses, from 4mg to SOOmg per wound per day for 8 days in vehicle. Vehicle control groups received SOmL of vehicle solution.
Animals are euthanized on day 8 with an intraperitoneal injection of sodium pentobarbital (300mg/kg). The wounds and surrounding skin are then harvested for histology. Tissue specimens are placed in 10% neutral buffered formalin in tissue cassettes between biopsy sponges for further processing.
Four groups of 10 animals each (5 with methylprednisolone and 5 without glucocorticoid) are evaluated: 1) Untreated group 2) Vehicle placebo control 3) treated groups.
Wound closure is analyzed by measuring the area in the vertical and horizontal axis and obtaining the total area of the wound. Closure is then estimated by establishing the differences between the initial wound area (day 0) and that of post treatment (day 8).
The wound area on day 1 is 64mm2, the corresponding size of the dermal punch.
Calculations are made using the following formula: --[Open area on day 8] - [Open area on day 1 ] / [Open area on day 1 Specimens are fixed in 10% buffered formalin and paraffin embedded blocks are sectioned perpendicular to the wound surface (5mm) and cut using an Olympus microtome. Routine hematoxylin-eosin (H&E) staining is performed on cross-sections of bisected wounds. Histologic examination of the wounds allows assessment of whether the healing process and the morphologic appearance of the repaired skin is improved by treatment with a polypeptide of the invention. A calibrated lens micrometer is used by a blinded observer to determine the distance of the wound gap.
Experimental data are analyzed using am unpaired t test. A p value of < 0.05 is considered significant.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 51: Lymphadema Animal Model The purpose of this experimental approach is to create an appropriate and consistent lymphedema model for testing the therapeutic effects of a polypeptide of the invention in lymphangiogenesis and re-establishment of the lymphatic circulatory system in the rat hind limb. Effectiveness is measured by swelling volume of the affected limb;
quantification of the amount of lymphatic vasculature, total blood plasma protein, and histopathology. Acute lymphedema is observed for 7-10 days. Perhaps more importantly, the chronic progress of the edema is followed for up to 3-4 weeks.
Prior to beginning surgery, blood sample is drawn for protein concentration analysis. Male rats weighing approximately ~350g are dosed with Pentobarbital.
Subsequently, the right legs are shaved from knee to hip. The shaved area is swabbed with gauze soaked in 70% EtOH. Blood is drawn for serum total protein testing.
Circumference and volumetric measurements are made prior to injecting dye into paws after marking 2 measurement levels (0.5 cm above heel, at mid-pt of dorsal paw). The intradermal dorsum ofboth right and left paws are injected with 0.05 ml of 1%
Evan's Blue. Circumference and volumetric measurements are then made following injection of dye into paws.
Using the knee joint as a landmark, a mid-leg inguinal incision is made circumferentially allowing the femoral vessels to be located. Forceps and hemostats are used to dissect and separate the skin flaps. After locating the femoral vessels, the lymphatic. vessel that runs along side and underneath the vessels) is located.
The main lymphatic vessels in this area are then electrically coagulated suture ligated.
Using a microscope, muscles in back of the leg (near the semitendinosis and adductors) are bluntly dissected. The popliteal lymph node is then located.
The 2 proximal and 2 distal lymphatic vessels and distal blood supply of the popliteal node are then and ligated by suturing. The popliteal lymph node, and any accompanying adipose tissue, is then removed by cutting connective tissues.
Care is taken to control any mild bleeding resulting from this procedure.
After lymphatics are occluded, the skin flaps are sealed by using liquid skin (Vetbond) (AJ , Buck). The separated skin edges are sealed to the underlying muscle tissue while leaving a gap of ~0.5 cm around the leg. Skin also may be anchored by suturing to underlying muscle when necessary.
To avoid infection, animals are housed individually with mesh (no bedding).
' Recovering animals are checked daily through the optimal edematous peak, which typically occurred by day 5-7. The plateau edematous peak are then observed.
To evaluate the intensity of the lymphedema, the circumference and volumes of 2 designated places on each paw before operation and daily for 7 days are measured. ' The effect plasma proteins on lymphedema is determined and whether protein analysis is a useful testing perimeter is also investigated. The weights of both control and edematous limbs are evaluated at 2 places. Analysis is performed in a blind manner.
Circumference Measurements: Under brief gas anesthetic to prevent limb movement, a cloth tape is used to measure limb circumference. Measurements are done at the ankle bone and dorsal paw by 2 different people then those 2 readings are averaged.
Readings are taken from both control and edematous limbs.
Volumetric Measurements: On the day of surgery, animals are anesthetized with Pentobarbital and are tested prior to surgery. For daily volumetrics animals are under brief halothane anesthetic (rapid immobilization and quick recovery), both legs are shaved and equally marked using waterproof marker on legs. Legs are first dipped in water, then dipped into instrument to each marked level then measured by Buxco edema software(Chen/Victor). Data is recorded by one person, while the other is dipping the limb to marked area.
Blood-plasma protein measurements: Blood is drawn, spun, and serum separated prior to surgery and then at conclusion for total protein and Ca2+ comparison.
Limb Weight Comparison: After drawing blood, the animal is prepared for tissue collection. The limbs are amputated using a quillitine, then both experimental and control legs are cut at the ligature and.weighed. A second weighing is done as the tibio-cacaneal joint is disarticulated and the foot is weighed.
Histological Preparations: The transverse muscle located behind the knee (popliteal) area is dissected and arranged in a metal mold, filled with freezeGel, dipped into cold methylbutane, placed into labeled sample bags at - 80EC until sectioning. Upon sectioning, the muscle is observed under fluorescent microscopy for lymphatics..
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the invention.
Example 52: Suppression of TNF alpha-induced adhesion molecule expression by a Polypeptide of the Invention The recruitment of lymphocytes to areas of inflammation and angiogenesis involves specific receptor-ligand interactions between cell surface adhesion molecules (CAMS) on lymphocytes and the vascular endothelium.. The adhesion process, in both normal and pathological settings, follows a mufti-step cascade that involves intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and endothelial leukocyte adhesion molecule-1 (E-selectin) expression on endothelial cells (EC). The expression of these molecules and others on the vascular endothelium determines the efficiency with which leukocytes may adhere to the local vasculature and extravasate into the local tissue during the development of an inflammatory response. The local concentration of cytokines and growth factor participate in the modulation of the expression of these CAMS.
Tumor necrosis factor alpha (TNF-a), a potent proinflammatory cytokine, is a stimulator of all three CAMs on endothelial cells and may be involved iri a wide variety of inflammatory responses, often resulting in a pathological outcome.
The potential of a polypeptide of the invention to mediate a suppression of TNF-a induced CAM expression can be examined. A modified ELISA assay which uses ECs as a solid phase absorbent is employed to measure the amount of CAM expression on TNF-a treated ECs when co-stimulated with a member of the FGF family of proteins.
To perform the experiment, human umbilical vein endothelial cell (HUVEC) cultures are obtained from pooled cord harvests and maintained in growth medium (EGM-2; Clonetics, San Diego, CA) supplemented with 10% FCS and 1%
penicillin/sfreptomycin in a 37 degree C humidified incubator containing 5%
C02.
HUVECs are seeded in 96-well plates at concentrations of 1 x 104 cells/well in EGM
medium at 37 degree C for 18-24 hrs or until confluent. The monolayers are subsequently washed 3 times with a serum-free solution of RPMI-1640 supplemented with 100 U/ml penicillin and 100 mg/ml streptomycin, and treated with a given cytokine and/or growth factors) for 24 h at 37 degree C. Following incubation, the cells are then evaluated for CAM expression.
Human Umbilical Vein Endothelial cells (HUVECs) are grown in a standard 96 well plate to confluence. Growth medium is removed from the cells and replaced with 90 u1 of 199 Medium (10% FBS). Samples for testing and positive or negative controls are added to the plate in triplicate. (in 10 u1 volumes). Plates are incubated at 37 degree C for either 5 h (selectin and integrin expression) or 24 h (integrin expression only). Plates are aspirated to remove medium and 100 ~l of 0.1% paraformaldehyde-PBS(with Ca++
and Mg++) is added to each well. Plates are held at 4°C for 30 min.
Fixative is then removed from the wells and wells are washed 1X with PBS(+Ca,Mg)+0.5% BSA and drained. Do not allow the wells to dry. Add 10 p1 of diluted primary antibody to the test and control wells. Anti-ICAM-1-Biotin, Anti-VCAM-1-Biotin and Anti-E-selectin-Biotin are used at a concentration of 10 ~ g/ml ( 1:10 dilution of 0.1 mg/ml stock antibody). Cells are incubated at 37°C for 30 min.
in a humidified environment. Wells are washed X3 with PBS(+Ca,Mg)+0.5% BSA.

Then add 20 ~l of diluted ExtrAvidin-Alkaline Phosphotase (1:5,000 dilution) to each well and incubated at 37°C for 30 min. Wells are washed X3 with PBS(+Ca,Mg)+0.5% BSA. 1 tablet of p-Nitrophenol Phosphate pNPP is dissolved in 5 ml of glycine buffer (pH 10.4). 100 w1 of pNPP substrate in glycine buffer is added to each test well. Standard wells in triplicate are prepared from the working dilution of the ExtrAvidin-Alkaline Phosphotase in glycine buffer: 1:5,000 (10°) >
10'°'S > 10'~ > 10'~~5. 5 p1 of each dilution is added to triplicate wells and the resulting AP content in each well is 5.50 ng, 1.74 ng, 0.55 ng, 0.18 ng. 100 ~l of pNNP reagent must then be added to each of the standard wells. The plate must be incubated at 37°C for 4h. A
volume of 50 p1 of 3M
NaOH is added to all wells. The results are quantified on a plate reader at 405 nm. The background subtraction option is used on blank wells filled with glycine buffer only. The template is set up to indicate the concentration of AP-conjugate in each standard well [
5.50 ng; 1.74 ng; 0.55 ng; 0.18 ng]. Results are indicated as amount of bound AP-conjugate in each sample.
The studies described in this example tested activity of a polypeptide of the invention. However, one skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), agonists, and/or antagonists of the 'invention.
Example 53: Assay for the Stimulation of Bone Marrow CD34+ Cell Proliferation This assay is based on the ability of human CD34+ to proliferate in the presence of hematopoietic growth factors and evaluates the ability of isolated polypeptides expressed in mammalian cells to stimulate proliferation of CD34+
cells.
It has been previously shown that most mature precursors will respond to only a single signal. More immature precursors require at least two signals to respond.
Therefore, to test the effect of polypeptides on hematopoietic activity of a wide range of progenitor cells, the assay contains a given polypeptide in the presence or absence of other hematopoietic growth factors. Isolated cells are cultured for 5 days in the presence of Stem Cell Factor (SCF) in combination with tested sample. SCF
alone has a very limited effect on the proliferation of bone marrow (BM) cells, acting in such conditions only as a "survival" factor. However; combined with any factor exhibiting stimulatory effect on these cells (e.g., IL-3), SCF will cause a synergistic effect. Therefore, if the tested polypeptide has a stimulatory effect on a hematopoietic progenitors, such activity can be easily detected. Since normal BM cells have a low level of cycling cells, it is likely that any inhibitory~effect of a given polypeptide, or agonists or antagonists thereof, might not be detected. Accordingly, assays for an inhibitory effect on progenitors is preferably tested in cells that are first subjected to in vitro stimulation with SCF+IL+3, and then contacted with the compound that is being evaluated for inhibition of such induced proliferation.
Briefly, CD34+ cells are isolated using methods known in the art. The cells are thawed and resuspended in medium (QBSF 60 serum-free medium with 1% L-glutamine (SOOmI) Quality Biological, Inc., Gaithersburg, MD Cat# 160-204-101 ).
After several gentle centrifugation steps at 200 x g, cells are allowed to rest for one hour. The cell count is adjusted to 2.5 x 105 cells/ml. During this time, 100 p,1 of sterile water is added to the peripheral wells of a 96-well plate. The cytokines that can be tested with a given polypeptide in this assay is rhSCF.(R&D Systems, Minneapolis, MN, Cat# 255-SC) at 50 ng/ml alone and.in combination with rhSCF
and rhIL-3 (R&D Systems, Minneapolis, MN, Cat# 203-ML) at 30 ng/ml. After one hour, 10 ~,l of prepared cytokines, SO p,1 SID (supernatants at 1:2 dilution =
50 p,1) and ~,l of diluted cells are added to the media which is already present in the wells to 20 allow for a final total volume of 100 p,1. The plates are then placed in a 37°C/5% COZ
incubator for five days.
Eighteen hours before the assay is harvested, 0.5 ~,Ci/well of [3H] Thymidine is added in a 10 ~1 volume to each well to determine the proliferation rate.
The experiment is terminated by harvesting the cells from each 96-well plate to a filtermat using the Tomtec Harvester 96. After harvesting, the filfermats are dried, trimmed and placed into OmniFilter assemblies consisting of one OmniFilter plate and one OmniFilter Tray. 60 ~1 Microscint is added to each well and the plate sealed with TopSeal-A press-on sealing film A bar code 15 sticker is affixed to the first plate for counting. The sealed plates is then loaded and the level of radioactivity determined via the Packard Top Count and the printed data collected for analysis. The level of radioactivity reflects the amount of cell proliferation.

The studies described in this example test the activity of a given polypeptide to stimulate bone marrow CD34+ cell proliferation. One skilled in the art could easilymodify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), antibodies, agonists, and/or antagonists and fragments and variants thereof.
As a nonlimiting example, potential antagonists tested in this assay would be expected to inhibit cell proliferation in the presence of cytokines and/or to increase the inhibition of cell proliferation in the presence of cytokines and a given polypeptide.
In contrast, potential agonists tested in this assay would be expected to enhance cell proliferation and/or to decrease the inhibition of cell proliferation in the presence of cytokines and a given polypeptide.
The ability of a gene to stimulate the proliferation of bone marrow CD34+
cells indicates that polynucleotides and polypeptides corresponding to the gene are useful for the diagnosis and treatment of disorders affecting the immune system and hematopoiesis. Representative uses are described in the "Immune Activity" and "Infectious Disease" sections above, and elsewhere herein.
Example 54: Assay for Extracellular Matrix Enhanced Cell.Resnonse (EMECR
The objective of the Extracellular Matrix Enhanced Cell Response (EMECR) assay is to identify gene products (e.g., isolated polypeptides) that act on the hematopoietic stem cells in the context of the extracellular matrix (ECM) induced signal.
Cells respond to the regulatory factors in the context of signals) received .
from the surrounding microenvironment. For example, fibroblasts, and endothelial and epithelial stem cells fail to replicate in the absence of signals from the ECM.
Hematopoietic stem cells can undergo self renewal in the bone marrow, but not in in vitro suspension culture. The ability of stem cells to undergo self renewal in vitro is dependent upon their interaction with the stromal cells and the ECM protein fibronectin (fn). Adhesion of cells to fn is mediated by the a5.(3~ and a4.~31 integrin receptors, which are expressed by human and mouse hematopoietic stem cells.
The factors) which integrate with the ECM environment and responsible for stimulating stem cell self renewal has not yet been identified. Discovery of such factors should be of great interest in gene therapy and bone marrow transplant applications Briefly, polystyrene, non tissue culture treated, 96-well plates are coated with fn fragment at a coating concentration of 0.2 pg/ cm2. Mouse bone marrow cells are plated (1,000 cells/well ) in 0.2 ml of serum-free medium. Cells cultured in the presence of IL-3 ( 5 ng/ml ) + SCF ( 50 ng/ml ) would serve as the positive control, conditions under which little self renewal but pronounced differentiation of the stem cells is to be expected. Gene products are tested with appropriate negative controls in the presence and absence of SCF(5.0 ng/ml), where test factor supernates represent 10% of the total assay volume. The plated cells are then allowed to grow by incubating in a low oxygen environment ( 5% CO2, 7% O2, and 88% Nz ) tissue culture incubator for 7 days. The number of proliferating cells within the wells is then quantitated by measuring thymidine incorporation into cellular DNA.
Verification of the positive hits in the assay will require phenotypic characterization of the cells, which can be accomplished by scaling up of the culture system and using appropriate antibody reagents against cell surface antigens and FACScan.
One skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), antibodies, agonists, and/or antagonists and fragments and variants thereof.
If a particular gene product is found to be a stimulator of hematopoietic progenitors, polynucleotides and polypeptides corresponding to the gene may be useful for the diagnosis and treatment of disorders affecting the immune system and hematopoiesis. Representative uses are described in the "Immune Activity" and "Infectious Disease" sections above, and elsewhere herein. The gene product may also be useful in the expansion of stem cells and committed progenitors of various blood lineages, and in the differentiation and/or proliferation of various cell types.
Additionally, the polynucleotides and/or polypeptides of the gene of interest and/or agonists and/or antagonists thereof, may also be employed to inhibit the proliferation and differentiation of hematopoietic cells and therefore may be employed to protect bone marrow stem cells from chemotherapeutic agents during chemotherapy. This antiproliferative effect may allow administration of higher doses of chemotherapeutic agents and, therefore, more effective chemotherapeutic treatment.

Moreover, polynucleotides and polypeptides corresponding to the gene of interest may also be useful for the treatment and diagnosis of hematopoietic related disorders such as, for example, anemia, pancytopenia, leukopenia, thrombocytopenia or leukemia since stromal cells are important in the production of cells of hematopoietic lineages. The uses include bone marrow cell ex-vivo culture, bone marrow transplantation, bone marrow reconstitution, radiotherapy or chemotherapy of neoplasia.
Example 55: Human Dermal Fibroblast and Aortic Smooth Muscle Cell Proliferation The polypeptide of interest is added to cultures of normal human dermal fibroblasts (NHDF) and human aortic smooth muscle cells (AoSMC) and two co-assays are performed with each sample. The first assay examines the effect of the polypeptide of interest on the proliferation of normal human dermal fibroblasts (NHDF) or aortic smooth muscle cells (AoSMC). Aberrant growth of fibroblasts or smooth muscle cells is a part of several pathological processes, including fibrosis, and restenosis. The second assay examines IL6 production by both NHDF and SMC. IL6 production is an indication of functional activation. Activated cells will have increased production of a number of cytokines and other factors, which can result in a proinflammatory or immunomodulatory outcome. Assays are run with and without co-TNFa stimulation, in order to check for costimulatory or. inhibitory activity.
Briefly, on day 1, 96-well black plates are set up with 1000 cells/well (NHDF) or 2000 cells/well (AoSMC) in 100 p,1 culture media. NHDF culture media contains:
Clonetics FB basal media, lmg/ml hFGF, 5mg/ml insulin, 50mg/ml gentamycin, 2%FBS, while AoSMC culture media contains Clonetics SM basal media, 0.5 ~g/ml hEGF, 5mg/ml insulin, 1 p,g/ml hFGF, 50mg/ml gentamycin, 50 p,g/ml Amphotericin B, 5%FBS. After incubation @ 37°C for at least 4-5 hours culture media is aspirated and replaced with growth arrest media. Growth arrest media for NHDF contains ~ fibroblast basal media, 50mg/ml gentamycin, 2% FBS, while growth arrest media for AoSMC contains SM basal media, 50mg/ml gentamycin, 50~.g/ml Amphotericin B, 0.4% FBS. Incubate at 37C until day 2.

On day 2, serial dilutions and templates of the polypeptide of interest are designed which should always include media controls and known-protein controls.
For both stimulation and inhibition experiments, proteins are diluted in growth arrest media. For inhibition experiments, TNFa is added to a final concentration of 2ng/ml (NHDF) or 5ng/ml (AoSMC). Then add 1/3 vol media containing controls or supernatants and incubate at 37C/5% COZ until day 5.
Transfer 60p1 from each well to another labeled 96-well plate, cover with a plate-sealer, and store at 4C until Day 6 (for IL6 ELISA). To the remaining 100 p1 in the cell culture plate, aseptically add Alamar Blue in an amount equal to 10%
of the culture volume (101). Return plates to incubator for 3 to 4 hours. Then measure fluorescence with excitation at 530nm and emission at 590nm using the CytoFluor.
This yields the growth stimulation/inhibition data.
On day 5, the IL6 ELISA is performed by coating a 96 well plate with 50-100 ul/well of Anti-Human IL6 Monoclonal antibody diluted in PBS, pH 7.4, incubate ON
at room temperature.
On day 6, empty the plates into the sink and blot on paper towels. Prepare Assay Buffer containing PBS with 4% BSA. Block the plates with 200 p.l/well of Pierce Super Block blocking buffer in PBS for 1-2 hr and then wash plates with wash buffer (PBS, 0.05% Tween-20).. Blot plates on paper towels. Then add 50 pl/well of diluted Anti-Human IL-6 Monoclonal, Biotin-labeled antibody at 0.50 mg/ml.
Make dilutions of IL-6 stock in media (30, 10, 3, 1, 0.3, 0 ng/ml). Add duplicate samples to top row of plate. Cover the plates and incubate for 2 hours at RT on shaker.
Wash plates with wash buffer and blot on paper towels. Dilute EU-labeled Streptavidin 1:1000 in Assay buffer, and add 100 pl/well. Cover the plate and incubate 1 h at RT. Wash plates with wash buffer. Blot on paper towels.
Add 100 ~ul/well of Enhancement Solution. Shake for 5 minutes. Read the plate on the Wallac DELFIA Fluorometer. Readings from triplicate samples in each assay were tabulated and averaged.
A positive result in this assay suggests AoSMC cell proliferation and that the gene product of interest may be involved in dermal fibroblast proliferation and/or smooth muscle cell proliferation. A positive result also suggests many.potential uses-of polypeptides, polynucleotides, agonists and/or antagonists of the gene/gene product of interest. For example, inflammation and immune responses, wound healing, and angiogenesis, as detailed throughout this specification. Particularly, polypeptides of the gene product and polynucleotides of the gene may be used in wound healing and dermal regeneration, as well as the promotion of vasculargenesis, both of the blood S vessels and lymphatics. The growth of vessels can be used in the treatment of, for example, cardiovascular diseases. Additionally, antagonists of polypeptides of the gene product and polynucleotides of the gene may be useful in treating diseases, disorders, and/or conditions which involve angiogenesis by acting as an anti-vascular (e.g., anti-angiogenesis). These diseases, disorders, and/or conditions are known in the art and/or are described herein, such as, for example, malignancies, solid tumors, benign tumors, for example hemangiomas, acoustic neuromas, neurofibromas, trachomas, and pyogenic granulomas; artheroscleric plaques; ocular angiogenic diseases, for example, diabetic retinopathy, retinopathy of prematurity, macular degeneration, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis, retinoblastoma, .uvietis and Pterygia (abnormal blood vessel growth) of the eye; rheumatoid arthritis; psoriasis; delayed wound healing; endometriosis;
vasculogenesis; granulations; hypertrophic scars (keloids); nonunion fractures; .
scleroderma; trachoma; vascular adhesions; myocardial angiogenesis; coronary collaterals; cerebral'collaterals; arteriovenous malformations; ischemic limb angiogenesis; Osler-Webber Syndrome; plaque neovascularization;
telangiectasia;
hemophiliac joints; angiofibroma; fibromuscular dysplasia; wound granulation;
Crohn's disease; and atherosclerosis. Moreover, antagonists of polypeptides of the gene product and polynucleotides of the gene may be useful in treating anti-hyperproliferative diseases and/or anti-inflammatory known in the art and/or described herein.
One skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), antibodies, agonists, and/or antagonists and fragments and variants thereof.
Example 56: Cellular Adhesion Molecule (CAM) Expression on Endothelial Cells The recruitment of lymphocytes to areas of inflammation and angiogenesis involves specific receptor-ligand interactions between cell surface adhesion molecules (CAMS) on lymphocytes and the vascular endothelium. The adhesion process, in both normal and pathological settings, follows a mufti-step cascade that involves intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and endothelial leukocyte adhesion molecule-1 (E-selectin) expression on endothelial cells (EC). The expression of these molecules and others on the vascular endothelium determines the efficiency with which leukocytes may adhere to the local vasculature and extravasate into the local tissue during the development of an inflammatory response. The local concentration of cytokines and growth factor .
participate in the modulation of the expression of these CAMS.
Briefly, endothelial cells (e.g., Human Umbilical Vein Endothelial cells (HUVECs)) are grown in a standard 96 well plate to confluence, growth medium is removed from the cells and replaced with 100-~l of 199 Medium (10% fetal bovine serum (FBS)). Samples for testing and positive or negative controls are added to the plate in triplicate (in 10 ~l volumes). Plates are then incubated at 37°C for either 5 h (selectin and integrin expression) or 24 h (integrin expression only). Plates are aspirated to remove medium and 100 ~l of 0.1% paraformaldehyde-PBS(with Ca++
and Mg++) is added to each well. Plates are held at 4°C for 30 min.
Fixative is removed from the wells and wells are washed 1X with PBS(+Ca,Mg) + 0.5% BSA
and drained. 10 ~1 of diluted primary antibody is added to the test and control wells.
Anti-ICAM-1-Biotin, Anti-VCAM-l-Biotin and Anti-E-selectin-Biotin are used at a .
concentration of 10 ~g/ml (1:10 dilution of 0.1 mg/ml stock antibody). Cells are incubated at 37°C for 30 min. in a humidified environment. Wells are washed three times with PBS(+Ca,Mg) + 0.5% BSA. 20 ~1 of diluted ExtrAvidin-Alkaline Phosphotase (1:5,000 dilution, refered to herein as the working dilution) are added to each well and incubated at 37°C for 30 min. Wells are washed three times with PBS(+Ca,Mg)+0.5% BSA. Dissolve 1 tablet of p-Nitrophenol Phosphate pNPP per 5 ml of glycine buffer (pH 10.4). 100 ~ l of pNPP substrate in glycine buffer is added to each~test well. Standard wells in triplicate are prepared from the working dilution of the ExtrAvidin-Alkaline Phosphotase in glycine buffer: 1:5,000 ( 10°) >
10-°'S > 10-' >
10-15. 5 ~l of each dilution is added to triplicate wells and the resulting AP
content in each, well_is 5.50 ng, 1.74 ng, 0.55 ng, 0.18 ng. 100 p1 of pNNP reagent is then added to each of the standard wells. The plate is incubated at 37°C for 4h. A volume of 50 p1 of 3M NaOH is added to all wells. The plate is read on a plate reader at 405 nm using the background subtraction option on blank wells filled with glycine buffer only. Additionally, the template is set up to indicate the concentration of AP-conjugate in each standard well [ 5.50 ng; 1.74 ng; 0.55 ng; 0.18 ng]. Results are indicated as amount of bound AP-conjugate in each sample.
Example 57: Alamar Blue Endothelial Cells Proliferation Assay This assay may be used to quantitatively determine protein mediated inhibition of bFGF-induced proliferation of Bovine Lymphatic Endothelial Cells (LECs), Bovine Aortic Endothelial Cells (BAECs) or Human Microvascular Uterine Myometrial Cells (UTMECs). This assay incorporates a fluorometric growth indicator based on detection of metabolic activity. A standard Alamar Blue Proliferation Assay is prepared in EGM-2MV with 10 ng /ml of bFGF added as a source of endothelial cell stimulation. This assay may be used with a variety of endothelial cells with slight changes in growth medium and cell concentration.
Dilutions of the protein batches to be tested are diluted as appropriate.
Serum-free medium (GIBCO SFM) without bFGF is used as a non-stimulated control and Angiostatin or TSP-1 are included as a known inhibitory controls.
Briefly, LEC, BAECs or UTMECs are seeded in growth media at a density of 5000 to 2000 cells/well in a 96 well plate and placed at 37-C overnight.
After.the overnight incubation of the cells, the growth media is removed and replaced with GIBCO EC-SFM. The cells are treated with the appropriate dilutions of the protein ~of interest or control protein samples) (prepared in SFM ) in triplicate wells with additional bFGF to a concentration of 10 ng/ ml. Once the cells have been treated with the samples, the plates) is/are placed back in the 37° C incubator for three days.
After three days 10 ml of stock alamar blue (Biosource Cat# DAL 1100) is added to each well and the plates) is/are placed back in the 37°C incubator for four hours. The plates) are then read at 530nm excitation and 590nm emission using the CytoFluor fluorescence reader. Direct output is, recorded in relative fluorescence units.

Alamar blue is an oxidation-reduction indicator that both fluoresces and changes color in response to chemical reduction of growth medium resulting from cell growth. As cells grow in culture, innate metabolic activity results in a chemical reduction of the immediate surrounding environment. Reduction related to growth . causes the indicator to change from oxidized (non-fluorescent blue) form to reduced (fluorescent red) form. i.e. stimulated proliferation will produce a stronger signal and inhibited proliferation will produce a weaker signal and the total signal is proportional to the total number of cells as well as their metabolic activity. The background level .
of activity is observed with the starvation medium alone. This is compared to the output observed from the positive control samples (bFGF in growth medium) and protein dilutions.
Example 58: Detection of Inhibition of a Mixed Lymphocyte Reaction This assay can be used to detect and evaluate inhibition of a Mixed Lymphocyte Reaction (MLR) by gene products (e.g., isolated polypeptides).
Inhibition of a MLR may be due to a direct effect on cell proliferation and viability, modulation of costimulatory molecules on interacting cells, modulation of adhesiveness between lymphocytes and accessory cells, or modulation of cytokine production by accessory cells. Multiple cells may be targeted by these polypeptides since the peripheral blood mononuclear fraction used in this assay includes T, B and natural killer lymphocytes, as well as monocytes and dendritic cells.
Polypeptides of interest found to inhibit the MLR may find application in diseases associated with lymphocyte and monocyte activation or proliferation.
These include, but are not limited to, diseases such as asthma, arthritis, diabetes, inflammatory skin conditions, psoriasis, eczema, systemic lupus erythematosus, multiple sclerosis, glomerulonephritis, inflammatory bowel disease, crohn's disease, ulcerative colitis, arteriosclerosis, cirrhosis, graft vs. host disease, host vs. graft disease, hepatitis, leukemia and lymphoma.
Briefly, PBMCs from human donors are purified by density gradient centrifugation using Lymphocyte Separation Medium (LSM~, density 1.0770 g/ml, l Organon Teknika Corporation, West Chester, PA). PBMCs from two donors are adjusted to 2 x 106 cells/ml in RPMI-1640 (Life Technologies, Grand Island, NY) supplemented with 10% FCS and 2 mM glutamine. PBMCs from a third donor is adjusted to 2 x 105 cells/ml. Fifty microliters of PBMCs from each donor is added to wells of a 96-well round bottom microtiter plate. Dilutions of test materials (50 ~l) is added in triplicate to microtiter wells. Test samples (of the protein of interest) are added for final dilution of 1:4; rhuIL-2 (R&D Systems, Minneapolis, MN, catalog number 202-IL) is added to a final concentration of 1 ~g/ml; anti-CD4 mAb (R&D
Systems, clone 34930.11, catalog number MAB379) is added to a final concentration of 10 pg/ml. Cells are cultured for 7-8 days at 37°C in 5% COZ, and 1 ~C of [3H]
thymidine is added to wells for the last 16 hrs of culture. Cells are harvested and thymidine incorporation determined using a Packard TopCount. Data is expressed as the mean and standard deviation of triplicate determinations.
Samples of the protein of interest are screened in separate experiments and compared to the negative control treatment, anti-CD4 mAb, which inhibits proliferation of lymphocytes and the positive control treatment, IL-2 (either as recombinant material or supernatant), which enhances proliferation of lymphocytes.
One skilled in the art could easily modify the exemplified studies to test the activity of polynucleotides (e.g., gene therapy), antibodies, agonists, and/or antagonists and fragments and variants thereof.
It will be clear that the invention may be practiced otherwise than as particularly described in the foregoing description and examples. Numerous modifications and variations of the present invention are possible in light of the above teachings and, therefore, are within the scope of the appended claims.
The entire disclosure of each document cited (including patents, patent applications, journal articles, abstracts, laboratory manuals, books, or other disclosures) in the Background of the Invention, Detailed Description, and Examples is hereby incorporated herein by reference. Further, the hard copy of the sequence listing submitted herewith and the corresponding computer readable form are both incorporated herein by reference in their entireties. Additionally, the contents of U.S.
provisional applications Serial Nos. 60/179,065, 60/180,628, 60/251,988, 60/226,279, and Serial No. unknown (Atty. Docket PS730PP2) filed January 5, 2001 are all hereby incorporated by reference in their entirety.

INDICATIONS
RELATING
TO A
DEPOSITED
MICROORGANISM
OR OTHER
BIOLOGICAL
MATERIAL
(PCT
Rule 136is) A. The indications made below relate to the deposited microorganism or other biological material referred to in the description in Table I on page 67.

B. IDENTIFICATION
OF DEPOSIT
Further deposits are identified on an additional sheet Name of depositary institution.
American Type Culture Collection Address of depositary institution (including postal code and country) University Boulevard Manassas, Virginia United States of America Date Accession of deposit Number 09 June PTA-2079 C. ADDITIONAL
INDICATIONS
(leave blank ijnot applicable) This information is continued on an additional sheet D. DESIGNATED
STATES
FOR
WHICH
INDICATIONS
ARE
MADE
(ijthe indications are not jor all designated States) Europe In respect of those designations in which a European Patent is sought a sample of the deposited microorganism will be made available until the publication of the mention of the grant of the European patent or until the date on which the application has been refused or withdrawn or is deemed to be withdrawn, only by the issue of such a sample to an expert nominated by the person requesting the sample (Rule 28(4) EPC).
Continued on additional sheets E. SEPARATE
FURNISHING
OF INDICATIONS
(!care blank iJeioi apptieabte~

The indications listed below will be submitted to the international Bureau later (specify the general nature ojthe indications e.g..
'Accession Number ojDeposit') For receiving For International Bureau Office use only use only 0 This ~ This sheet sheet was was received received with by the the International international Bureau application on:

Authorized Authorized officer officer Revised Forth PCT/RO/134 (January 2001) Pctro134ep.sollist ATCC Deposit No. PTA-2079 CANADA
The applicant requests that, until either a Canadian patent has been issued on the basis of an application or the application has been refused, or is abandoned and no longer subject to reinstatement, or is withdrawn, the Commissioner of Patents only authorizes the furnishing of a sample of the deposited biological material referred to in the application to an independent expert nominated by the Commissioner, the applicant must, by a written statement, inform the International Bureau accordingly before completion of technical preparations for publication of the international application. .
NORWAY
The applicant hereby requests that the application has been laid open to public inspection (by the Norwegian Patent Office), or has been finally decided upon by the Norwegian Patent Office without having been laid open inspection, the furnishing of a sample shall only be effected to an expert in the art. The request to this effect shall be filed by the applicant with the Norwegian Patent Office not later than at the time when the application is made available to the public under Sections 22 and 33(3) of the Norwegian Patents Act. If such a request has been filed by the applicant; any request made by a third party for the furnishing of a sample shall indicate the expert to be used. That expert may be any person entered on the list of recognized experts drawn up by the Norwegian Patent Office or any person approved by the applicant in the individual case.
AUSTRALIA
The applicant hereby gives notice that the furnishing of a sample of a microorganism shall only be effected prior to the grant of a patent, or prior to the lapsing, refusal or withdrawal of the application, to a person who is a skilled addressee without an interest~in the invention (Regulation 3.25(3) of the Australian Patents Regulations).
FINLAND
The applicant hereby requests that, until the application has been laid open to public inspection (by the National Board of Patents and Regulations), or has been finally decided upon by the National~$oard of Patents and Registration without having been laid open to public inspection, the furnishing of a sample shall only be effected to an expert in the art.

ATCC Deposit No.: PTA-2079 UNITED KINGDOM
The applicant hereby requests that the furnishing of a sample of a microorganism shall only be made available to an expert. The request to this effect must be filed by the applicant with the International Bureau before the. completion of the technical preparations for the international publication of the application.
DENMARK
The applicant hereby requests that, until the application has been laid open to public inspection (by the Danish Patent Office), or has been finally decided upon by the Danish Patent office without having been laid open to public inspection, the furnishing of a sample shall only be effected to an expert in the art. The request to this effect shall be filed by the applicant with the Danish Patent Office not later that at the time when the application is made available to the public under Sections 22 and 33(3) of the Danish Patents Act.
If such a request has been filed by the applicant, any request made by a third party for the furnishing of a sample shall indicate the expert to be used. That expert may be any person entered on a list of recognized experts drawn up by the Danish Patent Office or any person by the applicant in the individual case.
SWEDEN
The applicant hereby requests that, until the application has been laid open to public inspection (by the Swedish Patent Office), or has been finally decided upon by the Swedish Patent Office without having been laid open to public inspection, the furnishing of a sample shall only be effected to an expert in the art. The request to this effect shall be filed by the applicant with the International Bureau before the expiration of 16 months from the priority date (preferably on the Form PCT/RO/134 reproduced in annex Z of Volume I of the PCT
Applicant's Guide). If such a request has been filed by the applicant any request made by a third party for the furnishing of a sample shall indicate the expert to be used. That expert may be any person entered on a list of recognized experts drawn up by the Swedish Patent Office or any person approved by a applicant in the individual case.
NETHERLANDS
The applicant hereby requests that until the date of a grant of a Netherlands patent or until the date on which the application is refused or withdrawn or lapsed, the microorganism shall be made available as provided in the 31 F( 1 ) of the Patent Rules only by the issue of a sample to an expert. The request to this effect must be furnished by the applicant with the Netherlands Industrial Property Office before the date on which the application is made available to the public under Section 22C or Section 25 of the Patents Act of the Kingdom of the Netherlands, whichever of the two dates occurs earlier.

<110> Human Genome Sciences, Inc.
<120> 22 Human Secreted Proteins <130> PS730PCT
<150> 60/179,065 <151> 2000-O1-31 <150> 60/180,628 <151> 2000-02-04 <150> 60/251,988 <151> 2000-12-05 <150> 60/226,279 <151> 2000-OB-18 <150> Unassigned <151> 2001-O1-05 <160> 146 <170> PatentIn Ver. 2.0 <210> 1 <211> 733 <212> DNA
<213> Homo Sapiens <400>

gggatccggagcccaaatcttctgacaaaactcacacatgcccaccgtgcccagcacctg 60 aattcgagggtgcaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatga 120 tctcccggactcctgaggtcacatgcgtggtggtggacgtaagccacgaagaccctgagg 180 tcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcggg 240 aggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggact 300 ggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccaacccccatcg 360 agaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccc 420 catcccgggatgagctgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttct 480 atccaagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaaga 540 ccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtgg 600 acaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgc 660 acaaccactacacgcagaagagcctctccctgtctccgggtaaatgagtgcgacggccgc 720 gactctagaggat 733 <210> 2 <211> 5 <212> PRT
<213> Homo Sapiens <220>
<221> Site <222> (3) <223> Xaa equals any of the twenty naturally ocurring L-amino acids <400> 2 Trp Ser Xaa Trp Ser <210> 3 <211> 86 <212> DNA
<213> Artificial Sequence <220>
<221> Primer_Bind <223> Synthetic sequence with 4 tandem copies of the GAS binding site found in the IRF1 promoter (Rothman et al., Immunity 1:457-468 (1994)), 18 nucleotides complementary to the SV40 early promoter, and a Xho I
restriction site.
<400> 3 gcgcctcgag atttccccga aatctagatt tccccgaaat gatttccccg aaatgatttc 60 cccgaaatat ctgccatctc aattag 86 <210> 4 <211> 27 <212> DNA
<213> Artificial Sequence <220>
<221> Primer_Bind <223> Synthetic sequence complementary to the SV40 promter; includes a Hind III restriction site.
<400> 4 gcggcaagct ttttgcaaag cctaggc 27 <210> 5 <211> 271 <212> DNA
<213> Artificial Sequence <220>
<221> Protein Bind <223> Synthetic promoter for use in biological assays; includes GAS binding sites found in the IRF1 promoter (Rothman et al., Immunity 1:457-468 (1994) ) .
<400> 5 ctcgagattt ccccgaaatc tagatttccc cgaaatgatt tccccgaaat gatttccccg 60 aaatatctgc catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 120 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 180 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 240 ttttggaggc ctaggctttt gcaaaaagct t 271 <210> 6 <211> 32 <212> DNA
<213> Artificial Sequence <220>
<221> Primer_Bind <223> Synthetic primer complementary to human genomic EGR-1 promoter sequence (Sakamoto et al., Oncogene 6:867-871 (1991)); includes a Xho I
restriction site.
<400> 6 gcgctcgagg gatgacagcg atagaacccc gg 32 <210> 7 <211> 31 <212> DNA
<213> Artificial Sequence <220>
<221> Primer_Bind <223> Synthetic primer complementary to human genomic EGR-1 promoter sequence (Sakamoto et al., Oncogene 6:867-871 (1991)); includes a Hind III
restriction site.
<400> 7 gcgaagcttc gcgactcccc ggatccgcct c 31 <210> 8 <211> 12 <212> DNA
<213> Homo sapiens <400> 8 ggggactttc cc 12 <210> 9 <211> 73 <212> DNA
<213> Artificial Sequence <220>
<221> Primer_Bind <223> Synthetic primer with 4 tandem copies of the NF-KB binding site (GGGGACTTTCCC), 18 nucleotides complementary to the 5' end of the SV40 early promoter sequence, and a XhoI restriction site.
<400> 9 gcggcctcga ggggactttc ccggggactt tccggggact ttccgggact ttccatcctg 60 ccatctcaat tag 73 <210> 10 <211> 256 <212> DNA
<213> Artificial Sequence <220>
<221> Protein Bind <223> Synthetic promoter for use in biological assays; includes NF-KB
binding sites.
<400> 10 ctcgagggga ctttcccggg gactttccgg ggactttccg ggactttcca tctgccatct 60 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 120 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 180 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 240 cttttgcaaa aagctt 256 <210> 11 <211> 1631 <212> DNA
<213> Homo Sapiens <400>

ggcacgagcttgtcgtgtggctctgtgtcgacactgtgcgccaccatggccgtgactgcc 60 tgtcagggcttggggttcgtggtttcactgattgggattgcgggcatcattgctgccacc 120 tgcatggaccagtggagcacccaagacttgtacaacaaccccgtaacagctgttttcaac 180 taccaggggctgtggcgctcctgtgtccgagagagctctggcttcaccgagtgccggggc 240 tacttcaccctgctggggctgccagccatgctgcaggcagtgcgagccctgatgatcgta 300 ggcatcgtcctgggtgccattggcctcctggtatccatctttgccctgaaatgcatccgc 360 attggcagcatggaggactctgccaaagccaacatgacactgacctccgggatcatgttc 420 attgtctcaggtctttgtgcaattgctggagtgtctgtgtttgccaacatgctggtgact 480 aacttctggatgtccacagctaacatgtacaccggcatgggtgggatggtgcagactgtt 540 cagaccaggtacacatttggtgcggctctgttcgtgggctgggtcgctggaggcctcaca 600 ctaattgggggtgtgatgatgtgcatcgcctgccggggcctggcaccagaagaaaccaac 660 tacaaagccgtttcttatcatgcctcaggccacagtgttgcctacaagcctggaggcttc 720 aaggccagcactggctttgggtccaacaccaaaaacaagaagagatacgatggaggtgcc 780 cgcacagaggacgaggtacaatcttatccttccaagcacgactatgtgtaatgctctaag 840 acctctcagcacgggcggaagaaactcccggagagctcacccaaaaaacaaggagatccc 900 atctagatttcttcttgcttttgactcacagctggaagttagaaaagcctcgatttcatc 960 tttggagaggccaaatggtcttagcctcagtctctgtctctaaatattccaccataaaac 1020 agctgagttatttatgaattagaggctatagctcacattttcaatcctctatttcttttt 1080 ttaaatataactttctactctgatgagagaatgtggttttaatctctctctcacattttg 1140 atgatttagacagactccccctcttcctcctagtcaataaacccattgatgatctatttc 1200 ccagcttatccccaagaaaacttttgaaaggaaagagtagacccaaagatgttattttct 1260 gctgtttgaattttgtctccccacccccaacttggctagtaataaacacttactgaagaa 1320 gaagcaataagagaaagatatttgtaatctctccagcccatgatctcggttttcttacac 1380 tgtgatcttaaaagttaccaaaccaaagtcattttcagtttgaggcaaccaaacctttct 1440 actgctgttgacatcttcttattacagcaacaccattctaggagtttcctgagctctcca 1500 ctggagtcctctttctgtcgcgggtcagaaattgtccctagatgaatgagaaaattattt 1560 tttttaatttaagtcctaaatatagttaaaataaataatgttttagtaaaatgaaaaaaa 1620 aaaaaaaaaaa 1631 <210> 12 <211> 1699 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (1692) <223> n equals a,t,g, or c <220>

<221> SITE
<222> (1693) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1696) <223> n equals a,t,g, or c <400>

gaacggggsatcaccagcgacacctgckawttggactacytctgaascattctttccctt 60 yctagggctttgctggatcccgccsgccccgagatccaacagaacaaascagaraggaga 120 cttaatcccgtattctccagatgtacagatacacgcagagcgattcatcaactataacca 180 gacagtttctcgaatgcgaggtatctacacagctccctcgggtctggagtccacttgttt 240 ggttgtggcctatggtttggacatttaccaaactcgagtctacccatccaagcagtttga 300 cgttctgaaggatgactatgactacgtgttaatcagcagcgtcctctttggcctggtttt 360 tgccaccatgatcactaagagactggcacaggtgaagctcctgaatcgggcctggcgata 420 aagaacaaagactgtgcctaaaagtggagagccaggggagtgtgggtcagataagcagct 480 acagctgcagtttggtggattggtggagtatgtgtgtgtgtcagtgctcagctaagaact 540 gtagggaagatggatgaccttcacgcagaactccttttgggatatacatgatgcagaaag 600 gatcctacatggagagagacagaactctctcagctgacactctcagagattcctgatggg 660 ctttctcttgaagtccaaaggcgtctgcattgtttcctttctttgcccatccatgaatgt 720 tctgttttgttttgttttttttaataagaattccggctgatttttgtgaggcctgtttaa 780 attgactttactttgccttttgtgtttctcaattttatctagaaatctttctgacttttt 840 ccatctcttgcttcaaagtaagaggggaactctccttgccgactccaccttataggtaca 900 tttggtgttttgcactgggaagaaataggatccatccttagctgaggcttgaggactgat 960 ccagcctctcatggcttccctccaaagtaacttagggttgagggatctatatgtgatgtc 1020 aaaacttactttaaacctctagtttcgtgctgtcatttattaggctgggccaccaaatct 1080 ttgtttcaatttatcagaagccaagtgcatactagcgtcttgtttgttgcccattgccta 1140 tacttttcacctgagatgtgtgagttggggccttttaaaaactactgaattgtctgagcc 1200 ttgaagacatttccagggagaagagataatctctcatttcacccacaggctggtctaatc 1260 ataacctagttaaagatgtccttgtttaagaaccccattatttatttttagtttttaata 1320 taaattaacatgtgggtcattatatttctccttaaatgaggaaattttaaattttattga 1380 tctaacctttgaagctttaaaaaaggagaaagagggtaggggtgggaaactggcatactg 1440 tgtgtatagcactgccgattggctaggccactgtgtctctgctacaaattaaagaaatcc 1500 taaaagttttccttggtcatagagttggggaatgacagaatttttctttgttgtgaaatg 1560 tatgtacagagtagaccatctctagccctgtggtgaaagaggtacactcgaatgtttgca 1620 taaagcaagtgacaaatgaaaaaaaaaaaaaaaaaaaaaccccgggggggggcccggtaa 1680 cccatttggccnnaanggg 1699 <210> 13 <211> 950 <212> DNA
<213> Homo Sapiens <400>

cccacgcgtccggggtcaaccacgagagctggcgtgacagtggccagactgacggcctga 60 cctttggccacctgaagatggtgctgctgtgggcctctgtgctcttcccggcgcccgagg 120 actgggcagagctgcagggcgccgtgtaccgcctgctggtggtgctgctctgttgcctgg 180 ccacgcggaagctgccccacttcctccacccgcagcgcaacctgctgcagggcagcggcc 240 tggaccttggtgccatctaccagcgcgtggagggcttcgccagccagcccgaggcggccc 300 tgcgcatccacgccacccacctgggccgcagccccccgccgcgcatcggctccgggctca 360 aggcgctcctgcagctgccagccagtgaccccacttactgggccactgcctacttcgacg 420 tcctgctggacaagttccaggtcttcaacatccaggataaggaccggatctctgccatgc 480 agagcatcttccagaagaccaggactctgggaggcgaggagagctgagctgggccacctg 540 gtctcagccacctgttcttggctccccaacagactctgcactgcaccatgggaggctcct 600 gggatgtttggaagaagaaacgggcttctccttgagggggtagtggagggattttgtccc 660 cagcagtggcctctgagagtctttcagtgcctggtggggcagggcaggcctcttggagca 720 cctcctccctgggtcagggcctggatgcaggtgccaagctctccatgtggtgcatgttga 780 cccagccacgtggtgttgtcaagcaaacagcatcggcaggagacctggagctgaggactt 840 ggccctgcctgcactgtatgccattccttggtgatgaaatgctgtatatttggttttgaa 900 aaaatgaatgtgctgggtatacacagcagaaagggaaaaaaaaaaaaaaa 950 <210> 14 <211> 2624 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (2504) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (2549) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (2616) <223> n equals a,t,g, or c <400>

gctatttaggtgacactatagaaggtacgcctgcaggtaccggtccggaattcccgggtc,60 gacccac.gcgtccggcgggggcatcgccgcccgcgcccctctaagtgccgggccgcaagc 120 tccaccgcagccgcctgcaagcagcggcgcctcggccctcgacctgcgcgcaaagcctgt 180 gctggagccgtcctcccgcggcggggaccgggaccggggacccaagccaatcgaaagctc 240 caaccatggccatggggctcttccgcgtgtgtctggtggtggtgacggccatcatcaacc 300 acccgctgctgttcccgcgggagaacgccacagtccccgagaacgaggaggagatcatcc 360 gcaagatgcaggcgcaccaggagaagctgcagctggagcagttgcgcctggaggaggagg 420 tggctcggctggcggccgaaaaggaggcactggagcaggtggcggaggagggcaggcagc 480 agaacgagacacgcgtggcctgggacctctggagcaccctctgcatgatcctcttcctga 540 tgatcgaggtgtggcggcaggaccaccaggaggggccctcacctgagtgcctgggcggtg 600 aggaggatgagctgcctgggctggggggcgcccccttgcagggcctcaccctgcccaaca 660 aggccacgcttggccacttttatgagcgctgcatccggggggccacggccgatgcagccc 720 gtacccgggagttcctggaaggcttcgtggatgacttgctggaagccctgaggagcctct 780 gcaaccgggacaccgacatggaggtggaggacttcattggcgtggacagcatgtacgaga 840 actggcaggtggacaggccactgctgtgccaccttttcgtgcccttcacaccccccgagc 900 cctaccgcttccacccagagctctggtgctccggccgctcagtgcccctggatcgccagg 960 gctacggccagatcaaggtggtccgcgccgatggggacacattgagctgcatctgcggca 1020 agaccaagctcggggaagacatgctgtgtctcctgcacggcaggaacagcatggcgcctc 1080 cctgcggygacatggagaacctgctgtgtgccacagattccctgtacctggacacgatgc 1140 aggtcatgaagtggttccagacggccctcaccagagcctggaagggcatcgcccacaagt 1200 acgagttcgacctggcctttggccagctggacagcccggggtccctgaagatcaagttcc 1260 gttcagggaagttcatgcccttcaacctgattcctgtgatccagtgtgatgactcggacc 1320 tgtactttgtctcccaccttcccagggagccctctgagggcaccccagcctccagcacag 1380 actggctcctgtcctttgctgtctatgagcgacacttcctcaggacgacactaaaggcac 1440 tgcccgagggcgcctgccacctcagctgcctgcagatagcatccttcctgctctccaagc 1500 agagccgcctgaccggtcccagcgggctcagcagctaccacctgaagacggccctactgc 1560 acctcctactcctccggcaggccgccgactggaaggcggggcagctggacgctcgtctgc 1620 acgagttgctgtgcttcctggagaagagcttgctccagaagaagctccaccacttcttca 1680 tcggcaaccgcaaggtgcctgaggccatgggactccctgaggccgtgctcagggccgagc 1740 ccctcaacctcttccggcccttcgtcctgcagcgaagcctttaccgtaagacactggact 1800 ccttctatgagatgctcaagaatgccccagcgctcattagcgagtattccctacatgtcc 1860 cctcagaccagcctaccccaaaaagctgacgtcttttacagaatgtgggatcctcgagct 1920 aagatgagggcatccctcacgttcacacccctggtggcatctgccagccctgttctgggg 1980 acaaggcgggctttcgtgggagccgtgctcagcctgccaggaagccaagccctacagtgc 2040 agaggaaacagaatttcaacgggaagctggtttgcttcataccattgggatctgctggta 2100 aagctgttatttgggtttagggactgatcccttgcagtttacttctggatcaccatgaat 2160 ggccaagatggtggcagaacacgctgtggaccctgagttagagacaatgcaaatgttgga 2220 ttgggtgtaattctttttgaatcccagatccagtctgtacttgaatatgagcagaggatc 2280 tacaagaatgctgacagggaaccgtgttaagacccagcacccctattcccaggagcttct 2340 ggcctgaccatctgcagccaaagcactaacagggacagatatgggaatgtccacctttga 2400 tccgcatcctgcacaatagtggtcccaccatggctgccacttttttatactatttggaga 2460 aaagaccttgtataaattcgaggcccgaatgactaacgtctctntcacacggaaatgggt 2520 acttggtgggcatagagacaatgcaaatntttggattgggtgtaattcttttttgaatcc 2580 cagatccagtctgtacttggaatatgagccagaggntctacaag 2624 <210> 15 <211> 1959 <212> DNA
<213> Homo Sapiens <400> 15 ggcacgagcgtgatcatcgggatgctcgtgctcctgctggactttcttggcttggtgcac 60 ctgggccagctgctcatcttccacatctacctgaaggccaagaagatgaccacctttgag 120 tatctcattaataaccgcaaagaagagagttcaaaacatcaagcagtgaggaaagatcca 180 tacgtgcaaatggacaaaggagttctccagcaaggagctggcgccctgggctcatctgca 240 cagggagtcaaagccaagagctccctgctgattcacaagcacttatgtcacttctgcact 300 tcagtaaaccaggatggggattcgacggcacgggtgcatttgtgagtcccctcagtgtct 360 ctgcagcttctatgtgtggatgaatagtgaagccacatgaggcctggtctgaagcagaga 420 agatccgctcagcatcatgttgaatcccaagcccgcgtctccgtgggctccaggacaatc 480 ctatgaaagtgacaccgtcgttcattgttcacatcggggaggagaattccatctgaaaat 540 gcacgtgacttcactgacacccgcgtccgtggcacagccctgtgctgagctccacagacc 600 tacagtccatcgcctccccttggagtgggcccacagctgcagacagtgttgtccaaagaa 660 gatgctctgagtagatgaatggaacaaactggaaactccagaaacaagcccttctttaca 720 tttctaatgtttggcttaaatggaaggtgccaagagccaaggccaggaactcccggctga 780 tttgcaggcgcctgtgtcagttctccactcgtgtacacccagacgggggctcgatggcac 840 aggaagcagatgatgccccgagtatatctacacttgggctgcaacaagaaacaacagagc 900 ccatgaaaactgacagtgctgaaagtgaagactgagattcaggagctcaggtgcccctgt 960 gatccaggtcttctaccctgaaaccccaccctccatcaaggtcctgcctgtagagtctac 1020 cttgcaaagcctcctgctcctacccatgctacaggccaggaaccagagcccatcatctca 1080 gaggcccctggatgtccttcgaaggaaccaggaccctcagagcccagcatccatctctgt 1140 catcatcttcatcacacccaaagaagagccagccttgcaggagggtttacatctccagga 1200 agatgggctgccagcaactgcagaggatgcagccacctgcttaactgtgctgtccagcca 1260 gccagccagctgcagggcctcttgctgcttaagagctgatgggccgggcatgttggctca 1320 cacctgtgagcacagtactgggaaatgggagcacagtactaggaaatgggagcacagtac 1380 tgggaaatgggagcacagtactgggaaatgggggctcacagcactgcaaaatgggagcac 1440 agtattgggaaatgggagcacagtactgggaagtgggagcacagtactgagaagtgggag 1500 cacagtactgagaaatgggagtacactactgagaaatgggagcacagtactgggaaatgg 1560 gcatacagtactgggaaatgggcatacagtactgagaaatgggagcacagtactgggaaa 1620 tgggagcacagtactgggaaatgggagcccacagtactgggaaaggggagttcacagtac 1680 tcggaaatgggagcatagtactgggaaatgggagcacagtactgggaaatgggagcatag 1740 tactgggaaaccccagacctggattctgagtttttcagcctagcccagacttcttatctt 1800 agtagacaaaaagagtcaataccagagaaccagaggcatcctctgtattttaatgaactc 1860 tgcattttaatctgtttagtagtcattttttaaaagataatcagttttccaaatatatct 1920 ataagttactacgtgcaaaaaaaaaaaaaaaaaaaaaaa 1959 <210> 16 <211> 3024 <212> DNA
<213> Homo sapiens <400> 16 ccacgcgtccggagcgaggctgccctttcttcgcagcgtgatttattttcttcttttttt 60 ctgaactcttcttccagggagaggctagtggtaacaggccgagctggatggatgggtatg 120 gggagaggggcaggacgttcagccctgggattctggccgaccctcgccttccttctctgc 180 agcttccccgcagccacctccccgtgcaagatcctcaagtgcaactctgagttctggagc 240 gccacgtcgggcagccacgccccagcctcagacgacacccccgagttctgtgcagccttg 300 cgcagctacgccctgtgcacgcggcggacggcccgcacctgccggggtgacctggcctac 360 cactcggccgtccatggcatagaggacctcatgagccagcacaactgctccaaggatggc 420 cccacctcgcagccacgcctgcgcacgctcccaccggccggagacagccaggagcgctcg 480 gacagccccgagatctgccattacgagaagagctttcacaagcactcggccacccccaac 540 tacacgcactgtggcctcttcggggacccacacctcaggactttcaccgaccgcttccag 600 acctgcaaggtgcagggcgcctggccgctcatcgacaataattacctgaacgtgcaggtc 660 accaacacgcctgtgctgcccggctcagcggccactgccaccagcaagctcaccatcatc 720 ttcaagaacttccaggagtgtgtggaccagaaggtgtaccaggctgagatggacgagctc 780 ccggccgccttcgtggatggctctaagaacggtggggacaagcacggggccaacagcctg 840 aagatcactgagaaggtgtcaggccagcacgtggagatccaggccaagtacatcggcacc 900 accatcgtggtgcgccaggtgggccgctacctgacctttgccgtccgcatgccagaggaa 960 gtggtcaatgctgtggaggactgggacagccagggtctctacctctgcctgcggggctgc 1020 cccctcaaccagcagatcgacttccaggccttccacaccaatgctgagggcaccggtgcc 1080 cgcaggctggcagccgccagccctgcacccacagcccccgagaccttcccatacgagaca 1140 gccgtggccaagtgcaaggagaagctgccggtggaggacctgtactaccaggcctgcgtc 1200 ttcgacctcctcaccacgggcgacgtgaacttcacactggccgcctactacgcgttggag 1260 gatgtcaagatgctccactccaacaaagacaaactgcacctgtatgagaggactcgggac 1320 ctgccaggcagggcggctgcggggctgcccctggccccccggcccctcctgggcgccctg 1380 gtcccgctcctggccctgctccctgtgttctgctagacgcgtagatgtggagggaggcgc 1440 gggctccgtcctctcggcttccccatgtgtgggctgggaccgcccacggggtgcagatct 1500 cctggcgtgtccaccatggccccgcagaacgccagggaccgcctgctgccaagggctcag 1560 gcatggacccctccccttctagtgcacgtgacaaggttgtggtgactggtgccgtgatgt 1620 ttgacagtagagctgtgtgagagggagagcagctcccctcgccccgcccctgcagtgtga 1680 atgtgtgaaacatcccctcaggctgaagccccccacccccaccagagacacactgggaac 1740 cgtcagagtcagctccttccccctcgcaatgcactgaaaggcccggccgactgctgctcg 1800 ctgatccgtggggccccctgtgcccgccacacgcacgcacacactcttacacgagagcac 1860 actcgatccccctaggccagcggggacaccccagccacacagggaggcatccttggggct 1920 tggccccaggcagggcaaccccggggcgctgcttggcaccttagcagactgctggaacct 1980 tttggccagtaggtcgtgcccgcctggtgccttctggcctgtggcctccctgcccatgtt 2040 cacctggctgctgtgggtaccagtgcaggtcccggttttcaggcacctgctcagctgccc 2100 gtctctggcctgggcccctgccccttccaccctgtgcttagaaagtcgaagtgcttggtt 2160 ctaaatgtctaaacagagaagagatccttgacttctgttcctctccctcctgcagatgca 2220 agagctcctgggcaggggtgcctgggccccagggtgtggcaggagacccagtggatgggg 2280 ccagctggcctgccctgatcctctgcttcctcctcacaaccccaagagcccccagcccgg 2340 tccatccacgtctggagtctggggagaggagcagggtcttaggactctcagctctgagca 2400 tccctggcagggtcttcaacctctaatctcttcccttaagccctgtggccacacagccag 2460 gagagacttgccgctggctcccgcctcatttcagcccagggtgctcatccaggggcccag 2520 aacagtcccacctgtgctgctatgcccacagcacaaagccaggcttcactcccaaaagtg 2580 cagccaggccctggagggtgatcctgccagcagccctacagctccacaccctacccaccc 2640 atcggcagcccctctgctgttccccagggacctctcatacactggccaggaggctgcaga 2700 acgtgtgtctccccctccctccaagaggtcctgctccctctgccagaaccgtgtgtgggc 2760 gggtgggagggcgctcggggcccggcccctccctctccctgctggttttagttggtccct 2820 atgttggaagtaaaaagtgaagcactttattttggttgtgtttgctcacgttctgcttgg 2880 aagtggggacccctcactgcgtccacgtgtctgcgacctgtgtggagtgtcaccgcgtgt 2940 acatactgtaaattatttattaatggctaaatgcaagtaaagtttggtttttttgttatt 3000 ttcttttaaaaaaaaaaaaaaaaa 3024 <210> 17 <211> 1151 <212> DNA
<213> Homo Sapiens <400> , ggcacgaggagaacaggggcaatgacacatgctccttggcttcctagtcctgatcccttg60 gggttccttgatcctagggtcatctgacttagatccatcctcccttcctctaggcaccag120 gggccatggctggaggtggccacccctcagtcctgtacagattctgtacccactggctgg180 agacccccatgcagctgtctcctgctcttgctgcggggagactgaactcagggctctgct240 cactggctccctcccaatggaagccttctctgggcttcacagcattgaatattcctccag300 aacagcttgttaaatcaaattttttttttttttagatggagtcttgctctgtcgcccagg360 ctggagtgcagtggcacgatcttggctcactgcaaactccgcctcccgggtttaagcgat420 tctcctgcctcggcctcccgaatagctgggactacaggtgctaccacacccagtaatttt480 tgtatttttagtagaaacggggattcactatgttggccaggctagtctcgaactcctgac540 ctcatgatctgtaatcccagcactttgggaggccgaggcaggtggatcacctaagttcgg600 gagtttgagaccagcctgaccaacatggagaaaccccatctctactaaatatacaaaatt660 aggctgggcgcagtggctcacgcctgtaatcccatcactttgggaggctgagacaggtgg720 gtccaaggtcaggagttcaagaccagcctggccaagatggtgaaaccccatctctactaa780 aaacacacacacacacacacaaaaattagccgggtgtggtggtgggcacctgtaatccca840 gctactagggaggctgaggcagagaactgcttgaacccggaaggcggaggttgcagtgag900 ccaagatcgcgccactgcactccaacctggacaacagggcaagactccctcttgaaaata960 taaaaaataaatatacaaaattagccaggtatggtggcgcatgcctgtggtcccggctgc1020 tcgggaggctgaggcaggagaatcacttgaatccgggagggagaggttgcggtgggctga1080 gattgcgccattgcactccagcctaggcagcaagagggaaactccgtctcaaaaaaaaaa1140 aaaaaaaaaaa 1151 <210> 18 <211> 961 <212> DNA
<213> Homo Sapiens <400>

actagtggatccaaagaattttggcacgagcacaaactgggtggcttaaaccacagaaat 60 gtattgtcttacagttctgggggccagaagtccaaaatcaaggtgtcagcaggcccatgc 120 tccatttgaaggcactggggaggggtctgctccaggcctgtctcctggctactggtggct 180 ccatggcttgtgacagcatcactccaatcttcacacagcattctccccgtgtgcctctga 240 gccccagttttcccttacttcaagagactggggcttttgctccatcacctagactggagg 300 gcagtggcatgatcttggctcactgctgccttgaactcctgtgctcaagtgatcctcctg 360 cctcggccatccagaattgctgggactatagatgtgagccacgacacctggcttcaattt 420 cctctttctataaggatatcggtcgtattggattagggcccgtcctactccagtataacc 480 tcatcttagtgaattacatctgccacaactctgtttccaaataaactcacattctgaggt 540 acagggagttaggacctcaacacaggaattttgggggctaaggtaaggagaaaacaagaa 600 gagtttggtgtcacagaagtgagaatgtccggccaggtatggtggctcaagcctgtgatc 660 ctagcactttgggaggccgaggtgggcagatcacctgaggtgagcagttcaagaccagcc 720 tggccaacatggtgaaactctgtctctactaaaaatacaaaaattagctgggcgtggtgg 780 cagacacctataatcccagctatccgggaa.gctgaggcaggagaatcacttgaacccagg 840 agatggaggctgcagttgcagtgagccaagatcacaccactgcactccagcttgggtgac 900 agagggagatgccgtctcaaaaaaaaaaaaaaaaaaaaaaaaactcgagagtacttctag 960 a 961 <210> 19 <211> 1609 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (1468) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1567) <223> n equals a,t,g, or c <400>

cggacgcgtgggggccgagtctggactagtgtactcctctttgcctttgccactggggta 60 cgctccaacgggctggtcagtgttggcttcctcatgcattctcaatgccaaggctttttc 120 tcttctctaacgatgctgaatcctctgagacagctctttaagctgatggcctctctgttt 180 ctgtcggtgttcacacttggccttccctttgccctctttcagtattatgcctacacccaa 240 ttctgtctgccaggctcagcccgccccattcctgagcctttggtacagttagctgtagac 300 aagggctaccggattgcagagggaaatgaaccgccttggtgcttctgggatgttccacta 360 atatacagctatawcyaggatgtctactggaatgttggctttttgaaatactatgagctc 420 aagcaggtgcccaattttctactggctgcaccagtggctatactggttgcctgggcaact 480 tggacatacgtgaccactcacccttggctctgccttacacttgggctgcaaaggagcaag 540 aacaataagaccctagagaagcccgatcttggattcctcagtcctcaggtgtttgtgtac 600 gtggtccacgctgcagtgctgctgctgtttggaggtctgtgcatgcatgttcaggttctc 660 accaggtttttgggctcctccactcctattatgtactggtttccagctcacttgcttcag 720 gatcaagagccgctgttgagatccttaaagactgtgccttggaagcctcttgcagaggac 780 tccccaccaggacaaaaggtccccagaaatcctatcatgggacttttgtatcactggaaa 840 acctgttctccagtcacacgatacattctaggctacttcctgacttactggctcctggga 900 ctactcctacattgcaacttcctgccttggacatgacctggactctccagggacaggttg 960 gaagccaacttaacccaggggtctgaaagtaaaaatacacattggaactgcctctgctgc 1020 cctgggatcattactgtgtccattataatctttctctttctctttgaaagctggtcagga 1080 atgggagaagtgtcagacactagagagccccttctggtcctggctagggcaaattttaga 1140 caactattttctctgtaagtgaagattgtcgtattccaagtctaaaatacacctggatct 1200 gtctagtcaatcaacatagcagagacagtcttaaacctaccattgacctgtgtgtaaatt 1260 taaatgtcaatttattgaagtgtaaatttcatcaaaggcattagctgacaggctggtaac 1320 agtccacacaagatggtataggcctgaacagtgtagtggcagtaataaagtgggaccatt 1380 ttttccaaatgcgcatgttctgatgtttttatgatgtgaggatttattgaaaactcaagt 1440 tcgatactctcagtacatgaaaaaaaanaagaagaaaaaaaaaaagtttttgccgggtcc 1500 tttggttcttcctgggtctgggctcgcctagggacctcgggatggccgcctatcgagggc 1560 tcgcggnggcggtccggtgtttgcccagggtccaaggtctgtccgcttg 1609 <210> 20 <211> 1088 <212> DNA
<213> Homo sapiens <400>

ctactatagggaamgctkktacgcctgcaggtmccggtccggaattcccgggtcgaccca 60 cgcgtccggaggtggggggcagagtccccaggcagttgggattccggggagggaccacag 120 tgctgggaaccacagcactgacgccatacacaccaggggatgtttgctcttgcatggaaa 180 gtgattttcagtgtcatgctgcagaatcccatacgttatccttcagtccttggtataaag 240 tcctccctgctcagtagtcttgtgttggtgatggtttggggaaatgaaaagagtgggccc 300 tgccccacaccaaagtccaggaagggaagaaggagctgtccagcacaggtggggagggga 360 gaagaggggtcctattgggacccagaattcagactgagcaggaaatccaaccaggggttg 420 aggagggattacctgagcttgtatcacttcaacctgcattttagagacacattctaatct 480 cacagatgcagaatttggggatccctggacagctaacacttggggtgcaaatgtttcttg 540 ttgaatgagaggaactgggtgtagtaaatgaaattcttgcatggaagtctgaatacaact 600 ggtggccgcatcccgaccaacagacaagaggacaaactttcaaaaatgctgtgtcttctt 660 ctcatccttcttttagagtctgtctaccagaattccaccatctgaagtccccggggtgca 720 aatcctgtttgctgttaattcttccagctctcacgtatggtggcttgttgtgtcacgtgt 780 tttgcaattttgcattgcgcgttccatctttgtcagagctttatccgtgggaatcggtgt 840 gtctaggctgggagtccgtctctccctggggggtgagcttatacttccacgaggcaccag 900 aagacactccaaacccaagactgcttgaagtttagttcccatcttgcatgkagaataaat 960 tcagatccaaaatattcatgagaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaagggcggc 1020 cgctctagaggatccaagcttacgtacgcgtgcatgcgacgtcatagctcttctatagtg 1080 tcacctaa 1088 <210> 21 <211> 866 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (833) <223> n equals a,t,g, or c <400>

21 gaatacgccagctctaatacgactcactatagggaaagctggtacgcctgcaggtaccgg 60 tccggaattcccgggctcttgatctttcctggtcctggtacggggatgacctctctttta 120 gaggggcgaatggttctctgtgtctcctgcctcctccttcccctgctgttgttgctgaaa 180 cactttaatgggttgatgacaccatatttagctcataatgtatattgtcctatagaatat 240 atttcttttttcccatttcatgaaaaaaatatagaatatatttctatttggttcatcttt 300 gacagttttaagtttatctattcgaggctt.ttgtgtatttcccaaatttatgtgctttat 360 agagcatatactttgccccattagatgcagtggtattgcaatagttatcctcagggaggt 420 aggctgtgtgtgtgtgtgggtgtgagtgtatgtatttgtataaaagaaaatccaatatta 480 catgcatttggccgggcacggtggctcacatttgtaatctcagtgctttgggaggctgag 540 gtggacaggtcatttgaggtcaggagttcgagaccagcctggccaacatggtgaaactct 600 gtctctactaaaaattaaaaaattagccaggcgtggtggtgtacatctgtaatcccagct 660 acctgggaggctgaggcaggagaatctcttgaacctgggaggtggaagctgcagtgagcc 720 aagattgcaccactgcactccaggctgggggacagagcaagactccggtctcttcaaaaa 780 aaaaaaaaaaaaaaaaaaaaaaaaaaaaagggcggccgctctagaggatccangcttacg 840 tacgcgtgcatgcgacgtcatactct 866 <210> 22 <211> 1395 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (922) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (935) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1385) <223> n equals a,t,g, or c <400> 22 gaattcggcacgaggcggcgcgggacctgcagtcgccagggattccctccaggtgacgat 60 gctctggttctccggcgtcggggctctggctgagcgttactgccgccgctcgcctgggat 120 tacgtgctgcgtcttgctgctactcaattgctcgggggtccccatgtctctggcttcctc 180 cttcttgacaggttctgttgcaaaatgtgaaaatgaaggtgaagtcctccagattccatt 240 tatcacagacaacccttgcataatgtgtgtctgcttgaacaaggaagtgacatgtaagag 300 agagaagtgccccgtgctgtcccgagactgtgccctggccatcaagcagaggggagcctg 360 ttgtgaacagtgcaaaggttgcacctatgaaggaaatacctataacagctccttcaaatg 420 gcagagcccggctgagccttgtgttctacgccagtgccaggagggtgttgtcacagagtc 480 tggggtgcgctgtgttgktcattgtaaaaaccctttggagcatctgggaatgtgctgccc 540 cacatgtccaggctgtgtgtttgagggtgtgcagtatcaagaaggggaggaatttcagcc 600 agaaggaagcaaatgtacca.agtgttcctgcactggaggcaggacacaatgtgtgagaga 660 agtctgtcccattctctcctgtccccagcaccttagtcacatacccccaggacagtgctg 720 ccccaaatgtttgggtcagaggaaagtgtttgacctcccttttgggagctgcctctttcg 780 aagtgatgtttatgacaatggatcctcatttctgtacgataactgcacagcttgtacctg 840 cagggactctactgtggtttgcaagaggaagtgctcccaccctggtggctgtgaccaagg 900 ccaggagggctgttgtgaarantgcctcctacgantgcccccagaagacatcaaagtatg 960 caaatttggcaacaagattttccaggatggagagatgtggtcctctatcaattgtaccat 1020 ctgtgcttgtgtgaaaggcaggacggagtgtcscaataagcagtgcattcccatcagtag 1080 ctgcccacagggcaaaattctcaacagaaaaggatgctgtcctatttgcactgaaaagcc 1140 cggcgtttgcacggtgtttggagatccccactacaacacttttgacggtcggacatttaa 1200 ctttcaggggacgtgtcagtacgttttgacaaaagactgctcctcccctgcctcgccctt 1260 ccaggtgctggtgaagaacgacgcccgccggacacgctccttctcgtggaccaagtcggt 1320 ggarctggtgctgggcgaracagggtcagcctgcagcagcacctcaccgtgcgctggtac 1380 ggctngcgcatcgct 1395 <210> 23 <211> 3140 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (3) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (5) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (21) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1628) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1642) <223> n equals a~,t,g, or c <220>
<221> SITE

<222> (1656) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (2094) <223> n equals a,t,g, or c <400>

tgntncgagtcacccaggtgnagaatgaggagaaactgaaggagctagagcagatttgat 60 cccaaactctttctkgttttccttcttggacttatgctatttttttgtggagacttgctg 120 agcagaagtcaaattttctactactctactgggatgactgtgggaattgtggcctctctg 180 ctaatcatcatttttatactatctaagtttatgcctaagaaaagtcccatttacgtcatc 240 ctggtgggaggctggtctttttctctgtacctcattcaactagtttttaaaaatttacaa 300 gagatctggaggtgttactggcagtatcttttaagttatgtcctcacagttggattcatg 360 agttttgcagtatgttacaagtatgggcccttggagaatgaacgaagtatcaacctgctg 420 acctggaccttgcagctgatgggcctgtgtttcatgtattctggcatccagataccacat 480 attgcccttgccattatcatcattgctctttgtactaagaacctggaacaccctattcag 540 tggctgtacatcacctgcagaaaggtgtgtaagggagcagaaaagcctgttccccctcgt 600 ctcctgacagaagaagaatatcggatacaaggagaggtagaaacccgaaaggctttagag 660 gagctccgagaattttgtaacagtccagactgctctgcttggaagactgtttctcgaatc 720 cagtctccaaaaagatttgctgactttgtggaaggctcttcccacctcacgccaaatgaa 780 gtttctgtccatgagcaggagtatggattagggagcattattgcccaggatgaaatctat 840 gaggaagcatcctctgaggaggaggactcatattctcggtgtcctgctatcacacagaac 900 aactttctaacctaggtagtggtcagttatctttacgtggactggcttggtgccttggtc 960 catgttgcatgtgttgtgcaattgctttcaaccctttgaaacagagtgagatagataggg 1020 tagaaattctcctactgaaataagaggcctaaaaaggcctccctttggaaatgggaggtc 1080 tctatgggatccctgaggaaggagagtggataaagtagtgaatgctgggtagttcacttc 1140 ccattggttaagctaacagcccacttttatgtttccagagaaattggatggccacagcta 1200 gcatggcattctagctccttcttgaaagttgattcaatcatggcatttctgtcactggct 1260 ggctctccaaagtaagaactgttgttaagtgcaggaatgcttttagactataggctgcaa 1320 cttccagagagaaatccacaaatctgagcctccttcactccagcttttatttcrgtgact 1380 ttagaataattattgatttaactgttttgggaggaaaatagatttttattgttttgtttt 1440 ttaaatgaatgtcttttaaaaamcwtamcaaactcatgttccagaaccagcaagtgctcc 1500 agagtgacacaccccctaggcccctacatatttattaatatggattatccattaaagccc 1560 caggagctgttgttttaagctttgatttagttctcatacatatgatagaaagtcctattt 1620 gcctttangaacatgcctgtangctcttctgcaggntgagatgttctgggctttttatta 1680 tattcaactttcaattccatcttaaaaaacatttgtwttcttctcttcccattcttcctt 1740 accctgcctttgccctttcaggaagggtcagttcccttacctgtgaactatgtatgttca 1800 gagtagcattattcctgctagctaggagaagtcatcttgtttaggggatttggatgcttt 1860 ttatacgttctccattttcctgtcattgggtcatgttatctttgagttgctatgaaatca 1920 ggaaactgtctccttttcctttcccttcctttgtctacatgctctgtccattcctttcag 1980 ccttttctcaccacccatactcccccaaatctgggtaatttttaagccttgaaactatgt 2040 agtttcttgatacacaatttgtagttatgcagcagccacaatttgcattgccanggaaat 2100 argctccaggttatcttcatgcctctgggtgctcattcagctgtcaagtttccatgaact 2160 tacacttatttatgattgcgtttctgacctgagatgtatgctgcctgttattgcagtagc 2220 attagtttcagattcttttgccattgcaaagtaccccttataaaccagcaatgtcatctg 2280 tgaggaagcaaattctcaagtgtctgtcatttacttggttctttttctttgtggtcttca 2340 cccttataccctggaaaagtctgtaattaccttagccaggaagatagatggtcatggcaa 2400 gcgcacagcaccagacttactggctcaccaagatgatggaaaaaggcagatgatttttta 2460 aaaagccgtaatgactcctttagaccagccatttagcgtggtaattttgaaaggcctagc 2520 tccattgcagacttccaaagggtcagctctgagactgccctccaggtgggcagttgatta 2580 tttccaccagtgttttccagagccttaaactgtcctaagtgacaactacctcagttggca 2640 ggaaagagacatatagtagaaagtgaaaaatgagcagtatttgggcagatgctatgggtt 2700 acagttgaagggtaaaaggaactttacattgggaaacctttatacccttgtgaattatgt 2760 acatggtaaaatgttctctctctacaaagaactattaaaacttctgaaatatactatttt 2820 ttaccttatttatagaaattgagacctagcatatttaagcataagtttattttaaaaaat 2880 aattcaactcgtgcaagtggtctcaggattctctggagattttggtgcctcccctactta 2940 gggaggtgat agcttgccta waagggtgac ttttccygat catgtcttta tttcaatgag 3000 aaagcactgt gaaattgtga aagawtctcc tctttctctg tttaataaac ccccatgaaa 3060 tataaaaaaa aaaaaaaagg gcggccgctc tagaggatcc ctcgaggggc ccaagcttac 3120 gcgtgcatgc gacgtcatag 3140 <210> 24 <211> 2316 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (39) <223> n equals a,t,g, or c <400>

cgggggactctcaacgggaaagacttcctatagggaatnctggtacgcctgcaggtaccg 60 gtccggaattcccgggtcgacccacgcgtccggkgagagaagggggccttatttctcgga 120 aaccagccctgctgccctcttcccatggccacaggagcctccccagtgacggagttgtga 180 gattgggggtttctggtgtcctgacttccgtcccagggatcaccttgggtttcccacgtt 240 gcagagactaactgaaaggacatgagggctttaccctgggaatgctctgctggggcaggt 300 gggtgttagctgcgattctgtgttattttcccatcctcagaggtctgcggggtttccgag 360 aggctcgccgggatttctggcggggggctgagagcctggaggctgccctgacccacaacg 420 cagaggttcccaggcgccgggcccaggaggcagaagaggcaggagctgctttgaggacgg 480 ctcgagctgggtaccggggacgggcactggattatgccctgcagatcaacgtgattgagg 540 acaagaggaagtttgacatcatggartttgtgctgcgtttggtggaggcccagsctaccc 600 atttcagcaggccatgaggagctgagccggctgtcccagtatcgaaaggagctgggcgcc 660 cagttgcaccagctggtcttgaattcagcacgagagaagagggacatggagcagagacac 720 gtgctgctgaaacagaaggagctgggtggggaggagccagaaccaagcttaagagagggg 780 cctggtggcctggtgatggaaggacatctcttcaaacgggccagcaacgcatttaagacc 840 tggagcagacgctggttcaccattcagagcaaccaactggtttaccagaagaagtacaag 900 gaccctgtgactgtggtggtggatgaccttcgtctctgcacagtgaaactctgccctgac 960 tcagaaaggcggttctgctttgaggtggtgtccaccagcaagtcctgcctcctccaggct 1020 gactcagagcgcctcctgcagctgtgggtcagtgctgtgcagagcagcattgcttctgcc 1080 ttcagtcaggctcgccttgatgacagcccccggggtccaggccagggctcaggacacctg 1140 gccataggctctgctgccaccctgggctctggtggaatggccaggggaagggagcctggg 1200 ggagtcgggcacgtggtggcccaggtccagagtgtggatggcaatgcccagtgctgcgac 1260 tgccgggagccagccccggagtgggccagcatcaaccttggtgtcaccctctgcattcag 1320 tgttccggcatccacaggagccttggtgttcacttctccaaagtccggtctctgaccctt 1380 gactcatgggagccagaactagtgaagctcatgtgtgagctgggaaatgtcatcatcaac 1440 cagatctatgaggcccgcgtggaggccatggcagtgaagaaaccagggcccagctgctcc 1500 cggcaggagaaggaggcctggattcacgctaaatacgtggagaagaagttcctgaccaag 1560 ctgcctgagattcgagggcgaagaggtggccgggggcgcccaagggggcagcctcctgtg 1620 cccccaaagccttccatcaggccccggccagggagcttgagatccaagccagagcccccc 1680 tctgaggacctgggaagcctgcaccctggggccctactgtttcgagcgtctgggcatcct 1740 ccatctcttcccaccatggctgatgcccttgcccatggagctgatgtcaactgggtcaat 1800 gggggccaagataatgccacaccgctgatccaggccacagctgctaattctcttctggcc 1860 tgtgagtttctcctccagaacggggcgaacgtgaaccaagcggacagtgcgggccggggc 1920 ccgctgcaccacgcaaccattcttggccacacggggctcgcctgcctgttcctgaaacgg 1980 ggagctgatctgggggctcgagactctgaaggcagggaccctctgaccatcgccatggaa 2040 acagccaacgctgacatcgtcaccctgctacgactggcaaagatgagggaggctgaagcg 2100 gcccaggggcaggcaggagatgagacgtatcttgacatcttccgcgacttctccctcatg 2160 gcgtcagacgacccggagaagctgagccgtcgcagtcatgacctccacacgctgtgaccc 2220 gaggcccacggggcccgcgcctgcctcccttccccgccaccgggccctctgccattaaag 2280 cctccgtgcttcgctcttcaaaaaaaaaaaaaaaaa 2316 IS
<210> 25 <211> 2304 <212> DNA
<213> Homo Sapiens <400>

ctcgaggtcgacggtatcgataagcttgatatcgaattcggcacgagctggtactatcaa 60 aactcaacagtataaggtatggatttagtatacttattaaatgaaagatatcctataata 120 tgacctgtgaatggattaaataaaaggtatgtacagttataaaattccataayaacagac 180 atgcaatttatttttttattttttaacttcttgccttttaataatatctttgaatataga 240 tggtttcaattctccaataacaagatgtagagtggccaaatggtttttcatacatgtttc 300 cctagtccagacacccaagagtatgtccctctcttgggcataagaaaacctagagttata 360 tttagggcatcatgtgtggtgaatagttgaccaagctggcatgctcacaatgaccaaatt 420 atgtttgtcatattttaatccaatacattgatctactatgaactkgtaaataatctattt 480 gcyttctagacacattccmggaagaccagagaaactgacttttatccatatggaataatc 540 cactggcaaatttcttttaggacagaccaaagctgatctataacataatctttacatact 600 trgatccttcccatcagacgagaaagaacatctggggtgaaaggctccattagaagagca 660 atgtgggatggtsctgtsctgacaactggtagttcctgccctacgtctttgtagaggact 720 ggagcccagagcatgccatggtcaactcgtatgttgaagtgaagacctccttcgaggtgt 780 gcatgtgcttattacagatatgccacggtgctaagtaccccaggactmcatgaaaaaata 840 agaatagatgctgtatccatgatcatgaggcacacaggccagagcgccagctgggaatcg 900 cacagcctgggttcaaagcctggctgggccatgaccacctgaatgacctgaggaatggtc 960 tcaggcaaatttgtaaaaagtggagaccctgcctgccagggaggcatgtggtaagaggtg 1020 catccagtcaggtcagggcactgcgtcctctctttggaacctgcggagcaaggctggtgg 1080 cccttgaggccacggcagccatggagaaggcgggcctggctccaggtggcacagaggcac 1140 tggagaggccccggggaagcctggcgcgatctggctggtcctgcgctctgcttccaggtt 1200 ctggccctgtaacctgggggacagggccggccaaaacagggccactgggtgctagccagc 1260 acytgggccaggcgccaggtggaagggctctggcggatcagcytcgcacccccaacagcc 1320 ccacaggggggcccatccagggccacacacttgcccccaggagcaggatatccctgaggc 1380 tagagtccagctggaccggtggaagggtctcaccctttgccctttgactcctcttgtagg 1440 caccctcgctgggctcctaagcactcctccacacactggctctgtcaccagccccatagt 1500 gatgtcataaactcccagatgcccagtgtgcacccagccacagagaagtgggtgacttag 1560 gagyatcctctccgcttctgacccttagtttcgtctgtgcacaactcactcaaaatgggc 1620 aactcgctaagcgtattttgttcctggttctgccgcaggtcctggccatgccatcggcaa 1680 cctgctcgtcttgtccgtgaggccttcccagctggccgggctcaccctgcagctcctgca 1740 cctgtgcctgcccggggaattgtgggccgtttcccactcctcttcaaccgtcagcgacat 1800 yttgggccttyttttccagtcaggtgggacggcgcccctatgaggctgtgtcttatccct 1860 cggaacacgggcaccccacagagggtcctgcgtcctgtggtctggagccccccctcaagg 1920 aagaaacccgtgctgtctcctcacaactccataatgtttggacacctcagccccgtaagg 1980 atcccttgtctcagaggcaagtttaaccttcaacttccttcattagatgatcaggtgatc 2040 ccagccaggctcccgaagacggaggtgagcgcagaagagcccaaagaagcaacagaggtg 2100 aaagaccaggtagagacccaggggcaggaggacaataaaaggggcccctgtagcaatggg 2160 gaagcagcctccacctctaggcccctggagactcagggaaacctcacttcttcctggtac 2220 aatcccaggcccttggagggaaatgtccacctcaagagcttgacagaaaagaatcagact 2280 gacaaggcccaggtgcatgcagtc 2304 <210> 26 <211> 2346 <212> DNA
<213> Homo Sapiens <400> 26 gcccacgcgt ccgcccacgc gtccggtgcg ggtttcggtt ggaggactcg ttggggaggt 60 ggcctgcgct tgtagagact gcatccccga gacgatggcg gagggagata atcgcagcac 120 caacctgctg gctgcagaga ctgcaagtct ggaagaacag ctgcaaggat ggggagaagt 180 gatgctgatg gctgataaag tcctccgatg ggaaagagcc tggtttccac ctgccatcat 240 gggtgtggtt tctttggtgt ttctgattat ctactatcta gatccatctg t'tctgtccgg 300 cgtttcctgttttgttatgtttttgtgcttggctgactaccttgttcccattctagcgcc360 tagaatttttggctccaataaatggaccactgaacaacagcaaagattccatgaaatttg420 cagcaatctagtaaaaactcgacgcagagctgtgggttggtggaaacgcctcttcacact480 aaaggaagaaaaacctaagatgtacttcatgaccatgatcgtttcccttgctgcggttgc540 ttgggtgggacaacaagtccacaacctgcttctcacctacctgatagtgacttccttact600 attgcttcctggactaaaccaacatggaatcattttgaagtacattggaatggccaagag660 ggagataaacaaacttctcaaacaaaaagaaaagaaaaacgaatgattcatctgctttaa720 tcagtgtgattaatgcagcacccattgccccgggaaccgtttctgctgtactatctggat780 actaaaatgttacggaagtagctctttgttctccctcactctgcccttagttaatagaaa840 ttcagactcgccaagtaaggcttcgtgcatagtgtcttcatgtcgcgtatagttgagcgc900 gttcttagcagttggcttcatggacaactcattagtgttttgacttttcttacccagcgt960 taattgaattcttgcttttagacaacttcctttttgtagtggtgaaccttgccctttagt1020 acagttcaagtgaatctggataattgttcatctttgctttagcttagataccatgtagtg1080 gtctgtggctacaggaagctggttctgtctgcttccacagtctgcttaaaaaactgtctg1140 acttcgtgaatatagagaccaagtttaccacttctgatgaagagaccaattaagattcat1200 tcctcattctgtttctttccagtgggagaagagtccccatgaaataagatgaaactgatt1260 ccatgcactagtacatgtaggcttctcccttgtgcaaagcttagcaatttgtaggaaact1320 ttgatctttttgtccaagaaaaggaatgtctgacaggcttaagctttcgtccccttgcac1380 ttagactcgaagttagtaaatccttaaaggctttttaatagcagacttccaaaagattgc1440 atttaggatttctagcatgcttttaatttcagattttcagctgacattagctatagtata1500 cagtaggttaagactcatgtctatgactttcactctaagactggcaaaaggacagcagtc1560 ttctatgtttagtcaatattcatttcagtagaagataatcttatctaatttttgagacca1620 gaataagccttttaaggtaaacctcaaaattatcattttatggtaatactgaccatttta1680 gtcccctaggtttgacatgggagatagtgactacactggtgtctgacttttttcctagag1740 atttctccctgaaaaatacaagggctgttggtgagagcagacttgaggtgatgatagttg1800 gcctctggtctacaaagatttcataactccttggaaagcttcttataatcattcttaact1860 tcttggtagctagaaatttagagtagttgaaatctttaggaatgaacttctgagggccaa1920 aaaatgtgactgacgggaacaattcttaaactgattaactagctgtaatatagttttgtg1980 aatttattgcactgatgttgtaccttgtggtatatctgtccctattaaataagtgttgtt2040 ttctcctctttaatattgctgtgaacagtggtgcccattgtagcatatgtttgatttttt2100 tttattatttcataagaaaactacgttaattttaccttactttcattgtaaataagcctg2160 tcttcctatctggattttttgtgtgcatacatattctactgattaactacttttgcagtt2220 ttaatcctgtattatttcttctactttgttttgtgtaaaaggggaaaaaataaaaaaagc2280 tggaatcttaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaagggc2340 ggccgc 2346 <210> 27 <211> 1672 <212> DNA
<213> Homo Sapiens <400>

gaattcggcacgagtgaacctgtggggcaagggggtcttccctgggccccaagagtgcag 60 agatgcctgggtccgcagcctggcagggcagctgcagctttccccagggagcttccgccc 120 tgccaactctgagagggtggggctcctacttgttgctggctcccactggttctgtggagc 180 atgcagcaccagctgcacctcctcatatgctggggcaagggctctaagtccaacacatca 240 tgcctgggcccagtgctaagctgctccaacatgtggagtctggctctgctggtagtggct 300 ggctctatgggggtagcatacagctctgtggtgatgtacgttttgctctgggtgccgctc 360 ccacttccttcccacttccttccttcaggggcccctgaagcacagcccacaacttgggcc 420 cagtccccacactccgtgtgcaagtgtggcaccamcttgggcccagctaaacctcagggc 480 ccctctttgcctamtcctccatgcctgatcatgttgctgtcctgccgcaggcagctaggt 540 ctggccccatcawggtggctgccggggktgggttcccatggtggcgaactccggggatgc 600 tcccaggggtgggctccagggattgcccacctcaatatctgcacctgatgacccttggtc 660 tgtccctgttccccactgcagctgacatcatggtastgatcacttcamacaggctgccgc 720 tgtcctcacatacaatttataatttaaattagaaagaaacataaataaacgacataagat 780 tttgaatataatctatgtaagtaatttcaaaatatgttttattatcttctccatgctaga 840 gaacattttattagtttatattttaactaattttaaaaacgtaaacataaaattacatgt 900 gttgtcaagctaactcaatcgatgtttgagataaaaattaacctaagaccaaagtatatt 960 attttgatttccttagaggaaaagttatgcagtaattacttgaagagtaattctctctac 1020 mattagcattgcagatgcatccattataccactaaagtacacttaataaataatacttaa 1080 garryyaccagacaccatakggaaaaataaaaataaaaagtcattttgaaaaaagtttag 1140 aaagtaaatcattcctagaatgcttggaaaagtatgtgcctgtgtgccagtatttctgat 1200 cctaagagttatgaatatcttatgaatagttatgaatatcactgatagtgaacctgagtg 1260 taaatgcttgggcttgggktgcagatggactacagatacaaatctaaagtagtgtwccta 1320 gtagcactgtaactcagttaagtacttactttttcatgccctcagtatttttatctgcaa 1380 aataggaataaaatagctaatcatgttatcataagggttaagttatgagatataataact 1440 atgaagtacttaatgggtaaaggaaaaaataagcttagcagttgttgttgttgttgttgt 1500 tgacgaccttatttcaccatatacacttgttagtaactcaaaggggttctccaaagtgaa 1560 atatttaaaaaacgaatgagataaattattattttgttattagaaaatatatttagcaat 1620 ttgtctcactggatcataaaactaattgtggtatgagaacatttataaaaca 1672 <210> 28 <211> 1539 <212> DNA
<213> Homo Sapiens <400>

aaaaacaatgggatgtcatrggataaataatatgagtcaaattttgccagaaagttctgg 60 aaatgtaggacttggttctttttataagtgcttttaaccttttcagactcattaatttca 120 taaaccaatttaagcaattatttctatcagctacagatattttcttcacaaatataaagg 180 gtttgctcccctcccsatcctccagcctccaccaatttgtgcctgtaactgccacttgat 240 agctgctcagtaagactttgtttaaatatcaggttctagtgccaacaaatccagaagtca 300 gaaaaattctaagttacaatcgccagaatatctttcatgcattctattttactttattgt 360 tacagtttaaacaaattcagaaacgtggggtgtttctggctacttgagtaaaacgcactg 420 caaacgcaaagggaacaacgctgtaaacttaaatgatttctgaacagctcaggatcctcc 480 tggaggggatccttttcatctcacatattgattctgaacactatgttagcacctgctccc 540 agaaggtcccttgacactggagcaagtcagaatatcttaatatcatagcttatttaacag 600 caggaaakcaactttaacatagacttgaggtttaggtagatctggtattatctagaaccc 660 aggcttaaattttgcaaaaagtttccaaagagcataagaaagtaaaacagaggtaccaga 720 actgaagacatcattaacaaacaaaatatttccactcctaaccaaactgcaaccctttca 780 gtatagcctgaggtatttttaaacatttgtacaagctaaagatcattcacagtttccaaa 840 gctgctttttactgagatagcttcaaattttcacaatatgtgactttcagacatgttttc 900 atataccagagtctctgccctcagactcatggaaacatcagagatatttagtggcagaaa 960 atggttcttctagtgataagggttctatttcccaattattgtgctattcctaggatgtga 1020 ttaaaaaaatatgaaatttgcaatttgtataaaaggaaaggaaacttgcttaaaattcaa 1080 aaagcaagtccagtctggtggaggtatgtaagatattaaataatagagtcagattctttc 1140 ttgaacattccagatgaacttgacagcagagagttctggttttccaacattttctccatc 1200 ttttcttttccagtggttttactgatctctctttgcttcttactatggatccacgggata 1260 aaacaaagcacagcacctccaataaggggagggactccagcgaggtagaatgccacatca 1320 taggagcccagtttgtcacgaagtaaccctgcaatgggtgggccaacagtcatgggtata 1380 gacatgaatccgagcagaaatccaattgcttgggagacatcctgggcaccaactaactca 1440 aaggctatgggagccataatggaaatgaagcatccatcgaagagacccatgatgaggcac 1500 acagcaatgagggccccaaagatgctacacagaggaatc 1539 <210> 29 <211> 1077 <212> DNA
<213> Homo Sapiens <400> 29 ggcacgagcg gcacgagccc acagcctgca ggaccgaggc ccctgtgtga gggactgggg 60 gtgtgaggag ttgccaggag aagcagaccc tgccaggtgc ccatctcctc tcagcacccc 120 tccccccaac acacttcctg taatggcgtc tccaatgggg agcccattag tcctgcttag 180 ggcagcacctaagtactgtggctgataaatataacatgagcccatcgattagaatcctcc 240 tagtcctacagcagctcggctccctaatggctcccctcccctcagcacactgactaccca 300 cctctcaccccccaccccaggctgattttctggtttacttgaggctgtggggcaaacatg 360 agaggagcagaagtgaggtgatgggtgcaacacccagagcccagtcaaaggggccactgc 420 ttggagacaggctggactggtctgggccctgaggcagcctggaagcccctcggtcagtta 480 ggggcagctaggggaggagccgcctgtgagttcccggggccccggccctgcggtaggaca 540 tggagcaaggaaggcagagggcccctactttctgtgaggcgcaagacacagcacctgccc 600 ttgggggcccttgggggctctcctggtaactcctcagagaaacctgcagtgatgccagct 660 ctggccctgtcaccacgccctcttctccctggcatctcctgcccaagcccccgctgcacc 720 tgtcccagcccagccaaccctctctgtcttcatttcacccgtctccagggcgtccacatt 780 tcctcatcagatcagggttctgagggcaggggctgtgtctcctgcatcagcttagggccc 840 ctaaggggtagggactgcttccccagggtgggggctctggaagtcatgaggcttttcccc 900 ccaaaccaccaaatttggactgagaactgcatagggtagggtctgccgcaccccccgccc 960 ccaccataaggtaatcaaaccaattaggccttcagacttgttaaccatcagtgctgggtg 1020 gaagctgagacagcagagtgagacagctctgctacacctccccccagcctgaaaccc 1077 <210> 30 .

<211>

<212> DNA

<213> Homo Sapiens <220>

<221> SITE

<222> (11) <223> n equals or c a,t,g, <220>

<221> SITE

<222> (12) <223> n equals or c a,t,g, <220>

<221> SITE

<222> (15) <223> n equals or c a,t,g, <220>

<221> SITE

<222> (2782) <223> n equals or c a,t,g, <220>

<221> SITE

<222> (2848) <223> n equals or c a,t,g, <400> 30 ccttttagct nntgnaaaaggctatttaggtgacactatagaaggtacgcctgcaggtac 60 cggtccggaa ttcccgggtcgacccacgcgtccgcccacgcgtccgyyyacgcgtccgcc 120 cacgcgtccg ggctggccctgggcatgggccagggcctcaaagcctggccacggtaccgg 180 gttgtgggct ccgcagacgctgggcagtacaacctggagatcacagatgctgagctctct 240 gacgacgcct cttacgagtgccaggccacggaggccgccctgcgctctcggcgggccaaa 300 ctcaccgtgc tcatccccccagaggacaccaggattgacggaggccctgtgattctactg 360 caggcaggca ccccccacaacctcacatgccgggccttcaatgcgaagcctgctgccacc 420 atcatctggt tccgggacgggacgcagcaggagggcgctgtggccagcacggaattgctg 480 aaggatggga agagggagaccaccgtgagccaactgcttattaaccccacggacctggac 540 atagggcgtg tcttcacttgccgaagcatgaacgaagccatccctagtggcaaggagact 600 tccatcgagctggatgtgcaccaccctcctacagtgaccctgtccattgagccacagacg660 gtgcaggagggtgagcgtgttgtctttacctgccaggccacagccaaccccgagatcttg720 ggctacaggtgggccaaagggggtttcttgattgaagacgcccacgagagtcgctatgag780 acaaatgtggattattcctttttcacggagcctgtgtcttgtgaggttcacaacaaagtg840 ggaagcaccaatgtcagcactttagtaaatgtccactttgctccccggattgtagttgac900 cccaaacccacaaccacagacattggctctgatgtgacccttacctgtgtctgggttggg960 aatccccccctcactctcacctggaccaaaaaggactcaaatatggggcccaggcctcct1020 ggctccccacccgaggctgctctctctgcccaggtcctgagtaacagcaaccagctgctg1080 ctgaagtcggtgactcaggcagacgctggcacctacacctgccgggccatcgtgcctcga1140 atcggagtggctgagcgggaggtgccgctctatgtgaacgggccccccatcatctccagt1200 gaggcagtgcagtatgctgtgaggggtgacggtggcaaggtggagtgtttcattgggagc1260 acaccacccccagaccgcatagcatgggcctggaaggagaacttcttggaggtggggacc1320 ctggaacgctatacagtggagaggaccaactcaggcagtggggtgctatccacgctcacc1380 atcaacaatgtcatggaggccgactttcagactcactacaactgcaccgcctggaacagc1440 ttcgggccaggcacagccatcatccagctggaagagcgagaggtgttacctgtgggcatc1500 atagctggggccaccatcggcgcgagcatcctgctcatcttcttcttcatcgccttggta1560 ttcttcctctaccggcgccgcaaaggcagtcgcaaagacgtgaccctgaggaagctggat1620 atcaaggtggagacagtgaaccgagagccacttacgatgcattctgaccgggaggatgac1680 accgccagcgtctccacagcaacccgggtcatgaaggccatctactcgtcgtttaaggat1740 gatgtggatctgaagcaggacctgcgctgcgacaccatcgacacccgggaggagtatgag1800 atgaaggaccccaccaatggctactacaacgtgcgtgcccatgaagaccgcccgtcttcc1860 agggcagtgctctatgctgactaccgtgcccctggccctgcccgcttcgacggccgcccc1920 tcatcccgtctctcccactccagcggctatgcccagctcaacacctatagccggggccct1980 gcctctgactatggccctgagcccacaccccctggccctgctgccccagctggcactgac2040 acaaccagccagctgtcctacgagaactatgagaagttcaactcccatcccttccctggg2100 gcagctgggtaccccacctaccgactgggctacccccaggccccaccctctggcctggag2160 cggaccccatatgaggcgtatgaccccattggcaagtacgccacagccactcgattctcc2220 tacacctcccagcactcggactacggccagcgattccagcagcgcatgcagactcacgtg2280 taggggccagagcctggctggggcatctctgcggggcagaggagaaggctttcacagctg2340 ttccctgatattcaggggcattgctcattgctcccttctcggaccagccttcttcctccc2400 accatggcaggtggggagcaggtctcccagaaacaccccgtcccgaggatggtgctctgt2460 gcatgccccagcctcctgggcctgcccttccctcttcttcgggaggatgtgtctcttctg2520 acctgcactcttgcctgaccctagaatggggacagggaaagtgaaggttagggaaagcag2580 aggggggcactttttagcattccctttctatcccacccctctgatctcccataagtggaa2640 atgggggtacccagggatgggcaggctttggcctagggacatgaagtatgggagtgggtg2700 gctgwggcacagacaggtggaaaacgggatagcctggcagtccctctgttgctgcattcg2760 tgcctgggtgcctctcttcttnctcaggtactgcagaagggagcgaacagggtactgtcg2820 ctcttgctacagacaagcctggcactgnattcaaatcagcttcattactggatcaaatgc2880 agtaccttgctaccatgggaca 2902 <210> 31 <211> 2508 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (1074) <223> n equals a,t,g, or c <400> 31 ggcacgagca gcgcgcactt accaccatgc agatccaggt cgcggggctg ctgcagtttg 60 ccgtgcccct cttctccact gcagaggaag acctgcttgc aatccagctc ctgctgaact 120 cctcagagtc cagccttcac cagctgaccg ccatggtgga ctgccgaggg ctgcacaagg 180 attatctgga cgctcttgct ggcatctgct acgacggcct ccagggcttg ctgtaccttg 240 gcctcttctc cttcctggcc gccctcgcct tctccaccat gatctgtgca gggccaaggg 300 cctggaagca cttcaccacc agaaacagag actacgatga cattgatgat gatgacccct 360 ttaacccccaagcctggcgcatggcggctcacagtcccccgaggggacagcttcacagct 420 tctgcagctacagcagtggcctgggaagtcagaccagcctgcagcccccggcccagacca 480 tctccaacgcccctgtctccgagtacatgaaccaagccatgctctttggtaggaacccac 540 gctacgagaacgtgccactaatcgggagagcctcccctccgcctacgtactctcccagca 600 tgagagccacctacctgtctgtggcggatgagcacctgaggcactacgggaatcagtttc 660 cagcctaacagactttcgggggttcctgcctcctttttccgttctggtttttaattagtg 720 caaatacaagctgcgtttctttaatagaaaccaaaggcatctggagcccgagaggcctcc 780 tgctgtggcagaggagcagctgggattcccgaccaaagccccagggggtgcagaagactc 840 accacgcgggccagcctctctcttttgccctgctctccacaccagaaatgcccccaggtg 900 cttggctgcctcagaggtaccatccctgagctggctgcctggccctgctcacccctacgc 960 ctcgcccttgccaggaggggagtggcagtgaggagggggccaggtcaggcaccaccatca 1020 agagagctgtgtgttctctctggtcccacaacgatgactctgcctctttgggtncaggcc 1080 ccaggcccaaagagcccagacgacccctctgtcctcgttccctgtcctcgttccctgcag 1140 gtaacatgagaagggctgatcaggagatgctctttaagaagttcgcacccctgctgacac 1200 cagaacagcccaaatcagagttcccagggccagacaggctcttcctgggccacagagggg 1260 aggcatcaggaaagctctgcagtggggggctggtggctccggggctgggggatcacaggc 1320 tggtgaaccccggtgggaacagaggtgaaagcctgccacattccgcctgtctccctaacc 1380 ctccattgcc~tcgcctctattccagaatcaatgctgcagaatgtgttagctgcagatagg 1440 catggtctcaggtatgaacagacactttgaaacgactttaggtctttcttttctccagtg 1500 ttttaaacatgttgattatccaaagaattgaaactcctagcacatccagtttttacaaca 1560 gatttgcagctcattccttaccctggttaggtcactacttttgcagattttgctggcact 1620 gatctggagatctgcagatctggaggagacgggaaggagtcgattcttaaataaggatca 1680 gtgaggcatcctgtcccaagctactgtttggtggggatctgggttcatctcacccacaga 1740 gggaggatctttaagaggagaaaaaagccaagagggaaagccagagttccctgttctagg 1800 ggactagccaaatgcctacatcagctgtcccctccctgttgtctccaagtaagtttgcca 1860 gaaaaggttttagcaaagtgctacaactgtgtctttataggaggataggcctctgccctg 1920 ccccacccccaccacctgtccccacccagtgtcccaggccacaggagcttattggccagg 1980 agggaataatgtcccccaatactgcctgttgagggaccagagttggggtctttggtgctt 2040 ccaacctcctgccaacctggagttcacaacaccagagccccacggcctcgcacactgaag 2100 caggggcgtgcggtgactcggtgcttctgttttggaagaaccacctgtcatcaaaacatg 2160 gacagcagggtgttctcagctcccagcgaagcctccacaacagaatggggccacagggca 2220 gccgggactccctgtctcacctacattaacccatgcatactgtatgccataaactcactt 2280 tggtatatccgcgtcacatgcagagaggaactctgcgacgtcaaagtgttgcttcttaaa 2340 gtttcattattggcaactagagggttgtttttaatgcatggaaactaaacagattcctcg 2400 gggagttcctgaaggaaccaggtgggcaaacctttgcttatatacatgcggcctcacctg 2460 gaagagaaataaaccacttgtactaaaaaaaaaaaaaaaaaaaaaaaa 2508 <210> 32 <211> 2114 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (2108) <223> n equals a,t,g, or c <400>

ggctaggtggcgcgcgggtccggcgggcggttggcttgagcgggaccggagctgaggcag 60 gaagagccggcgccatggtggagaaggaggaggctggcggcggcattagcgaggaggagg 120 cggcacagtatgaccggcagatccgcctgtggggactggaggcccagaaacggctgcggg 180 cctctcgggtgcttcttgtcggcttgaaaggacttggggctgaaattgccaagaatctca 240 tcttggcaggagtgaaaggactgaccatgctggatcacgaacaggtaactccagaagatc 300 ccggagctcagttcttgattcgtactgggtctgttggccgaaatagggctgaagcctctt 360 tggagcgagctcagaatctcaaccccatggtggatgtgaaggtggacactgaggatatag 420 agaagaaaccagagtcatttttcactcaattcgatgctgtgtgtctgacttgctgctcca 480 gggatgtcatagttaaagttgaccagatctgtcacaaaaatagcatcaagttctttacag 540 gagatgtttttggctaccatggatacacatttgccaatctaggagagcatgagtttgtag 600 aggagaaaactaaagttgccaaagttagccaaggagtagaagatgggcccgacaccaaga 660 gagcaaaacttgattcttctgagacaacgatggtcaaaaagaaggtggtcttctgccctg 720 ttaaagaagccctggaggtggactggagcagtgagaaagcaaaggctgctctgaagcgca 780 cgacctccgactactttctccttcaagtgctcttaaagttccgtacagataaaggaagag 840 atcccagttctgatacatatgaggaagattctgagttgttgctccagatacgaaatgatg 900 tgcttgactcactgggtattagtcctgacctgcttcctgaggactttgtcaggtactgct 960 tctccgagatggccccagtgtgtgcggtggttggagggattttggcacaggaaattgtga 1020 aggccctgtctcagcgggaccctcctcacaacaacttcttcttcttcgatggcatgaagg 1080 ggaatgggattgtggagtgccttggccccaagtgaactcaagatttggcagccccagaga 1140 tgccaactgcagcatgcccacctgtattccctgtccccttccttcatgaaggcatctcca 1200 ggcaaggaaaactgaagtcattggcccgatacaaaacatttcctgcaacgaaggaggtgg 1260 tgccgacgtgctgcttcccatcaccagcagctgctcgacaaggggcgcagggtggctgtc 1320 tttgttccagcactgttcaggctgcctgtcatcccgggcctgccagctcccctgagtgat 1380 gagcacttccaagcacccctctgccctttctctgtccttatgctgtcccggcctcgccag 1440 ccctctggggcattgtgggagatgcctgccaggaatgagcaagctctgttgctcgggagc 1500 ctcttgtcaccttcttggacttattccccacctgataccttatagagaaaagtgtgaatt 1560 caggtggagagtaggcccaggccccatgaggcaccagtggaagcacagctccaagttcag 1620 acaggtgcccttagagaggaaaaccatgacaggcaaatgcatttcctctggagtttgaga 1680 ccctgacaaacaacaggtggcatctggtgtgctgttcttgagttttcgtttaggattagt 1740 tgagttccagctgggttttgggagaaaggagatgctaccaagtcttggatgttagggcga 1800 gaccctgcaagttgagtattagagagcttgtctttcaaggcaggttcctggggcttcagg 1860 gctaggagggaggagcctgcccttttaacagaaccccagtcacatgcggctcaagtcact 1920 cagaggctgttgcatttcagggctatgttggtcctttgtttacctcctaaaccacagctg 1980 tttgtgtttcacatatgttgtgaattttccttggttctttttaaaggaatgataataaag 2040 ttacttgctttaggaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 2100 aaaaaaanaaaggg 2114 <210> 33 <211> 2304 <212> DNA

<213> Homoapiens S

<220>

<221> SITE

<222> (2273) <223> n ls a,t,g, equa or c <220>

<221> SITE

<222> (2278) <223> n ls a,t,g, equa or c <400> 33 atgaatacgccagctcgaaattaaccctcactaaagggaacaaaagctggagctccaccg 60 cggtggcggccgctctagaactagtggatcccccgggctgcaggaattcggcacgagctt 120 gtcgtgtggctctgtgtcgacactgtgcgccaccatggccgtgactgcctgtcagggctt 180 ggggttcgtggtttcactgattgggattgcgggcatcattgctgccacctgcatggacca 240 gtggagcacccaagacttgtacaacaaccccgtaacagctgttttcaactaccaggggct 300 gtggcgctcctgtgtccgagagagctctggcttcaccgagtgccggggctacttcaccct 360 gctggggctgccagccatgctgcaggcagtgcgagccctgatgatcgtaggcatcgtcct 420 gggtgccattggcctcctggtatccatctttgccctgaaatgcatccgcattggcagcat 480 ggaggactctgccaaagccaacatgacactgacctccgggatcatgktcattgkctcagg 540 tctttgtgcaattgctggagtgtctgtgtttgccaacatgctggtgactaacttctggat 600 gtccacagctaacatgtacaccggcatgggkgggatggtgcagactgttcagaccaggta 660 cacatttggtgcggctctgttcgtgggctgggtcgctggaggcctcacactaattggggg 720 tgtgatgatgtgcatcgcctgccggggcctggcaccagaagaaaccaactacaaagccgt 780

22 ttcttatcatgcctcaggccacagtgttgcctacaagcctggaggcttcaaggccagcac840 tggctttgggtccaacaccaaaaacaagaagagatacgatggaggtgcccgcacagagga900 cgaggtacaatcttatccttccaagcacgactatgtgtaatgctctaagacctctcagca960 cgggcggaagaaactcccggagagctcacccaaaaaacaaggagatcccatctagatttc1020 ttcttgcttttgactcacagctggaagttagaaaagcctcgatttcatctttggagaggc1080 caaatggtcttagcctcagtctctgtctctaaatattccaccataaaacagctgagttat1140 ttatgaattagaggctatagctcacattttcaatcctctatttctttttttaaatataac1200 tttctactctgatgagagaatgtggttttaatctctctctcacattttgatgatttagac1260 agactccccctcttcctcctagtcaataaacccattgatgatctatttcccagcttatcc1320 ccaagaaaacttttgaaaggaaagagtagacccaaagatgttattttctgctgtttgaat1380 tttgtctccccacccccaacttggctagtaataaacacttactgaagaagaagcaataag1440 agaaagatatttgtaatctctccagcccatgatctcggttttcttacactgtgatcttaa1500 aagttaccaaaccaaagtcattttcagtttgaggcaaccaaacctttctactgctgttga1560 catcttcttattacagcaacaccattctaggagtttcctgagctctccactggagtcctc1620 tttctgtcgcgggtcagaaattgtccctagatgaatgagaaaattattttttttaattta1680 agtcctaaatatagttaaaataaataatgttttagtaaaatgaaaaaaaaaaaaaaaaaa1740 ctcgagggggggcccggtacccaattcgccctatagtgagtcgtattacaattcactggc1800 cgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgc1860 agcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttc1920 ccaacagttgcgcagcctgaatggcgaatggcaaattgtaagcgttaatattttgttaaa1980 attcgcgttaaatttttgttaaatcagctcattttttaaccaataggccgaaatcggcaa2040 aatcccttataaatcaaaagaatagaccgagatagggttgagtgttgttccagtttggaa2100 caagagtccactattaaagaacgkggactccaacgtcaaagggcgaaaaaccgtctatca2160 gggcgatggcccactacgtgaaccattaccctaatcaaggttttttggggtcgaggtgcc2220 gtaaagcactaaatcggaaccctaaagggagccccgattaaagcttgacgggnaaagncg2280 gcaacgtgcgagaaggaaggagaa 2304 <210>34 <211>1323 <212>DNA

<213>Homo Sapiens <220>

<221>SITE

<222>(1133) <223>n equalsa,t,g,or c <220>

<221>SITE

<222>(1183) <223>n equalsa,t,g,or c <220>

<221>SITE

<222>(1202) <223>n equalsa,t,g,or c <220>

<221>SITE

<222>(1282) <223>n equalsa,t,g,or c <220>

<221>SITE

<222>(1302) <223>n equalsa,t,g,or c

23 <220>
<221> SITE
<222> (1321) <223> n equals a,t,g, or c <400>

gtgcatgcgctcgcatcatggcggctgagtgggcttctcgtttctggctttgggctacgc 60 tgctgattcctgcggccgcggtctacgaagaccaagtgggcaagtttgattggagacagc 120 aatatgttgggaaggtcaagtttgcctccttggaattttcccctggatccaagaagttgg 180 ttgtagccacagagaagaatgtgattgcagcattaaattcccgaactggggagatcttgt 240 ggcgccatgttgacaagggcacggcagaaggggctgtggatgccatgctgctgcacggac 300 aggatgtgatcactgtgtccaatggaggccgaatcatgcgttcctgggagactaacatcg 360 ggggcctgaactgggagataaccctggaca.gtggcagtttccaggcacttgggctggttg 420 gcctgcaggagtctgtaaggtacatcgcagtcctgaagaagactacacttgccctccatc 480 acctctccagtgggcacctcaagtgggtggaacatctcccagaaagtgacagcatccact 540 accagatggtgtattcttacggctctggggtggtgtgggccctcggagttgttcccttca 600 gccatgtgaacattgtcaagtttaatgtggaagatggagagattgttcagcaggttaggg 660 tttcaactccgtggctgcagcacctgtctggagcctgtggtgtggtggatgargctgtcc 720 tggtgtgtcctgacccgagctcacgttccctccaaactttggctctggagacggaatggg 780 agttgagacagatcccactgcagtctctcgacttagaatttggaagtggattccaacccc 840 gggtcctgcctacccagcccaacccagtggacgcttcccgggcccagttcttcctgcact 900 tgtccccaagccactatgctctgctgcagtaccattatggaacgctgagtttgcttaaaa 960 acttcccacagactgccctagtgagctttgccaccactggggagaagacggtggctgcag 1020 tcatggcctgtcggaatgaagtgcagaaaactagcagttctgaagatgggtcaatggggg 1080 arcttttcggagaagtctaagttcaaaggactcttctggcttgcttcaatcanacctaca 1140 ccatttaacctaatacctcgtggaagacaggtcggcgggctgnttggacaccacgataac 1200 cnttaagcctggaacagaaccgggacttcggcctgaaccggctggattttccagggggtt 1260 cttgaaaaaaggatgacttcangggcttaccgggcttttggnccaaaccagaaggatcat 1320 ngg 1323 <210> 35 <211> 2737 <212> DNA

<213> Homo Sapiens <220>

<221> SITE

<222> (2730) <223> n equals or c a,t,g, <220>

<221> SITE

<222> (2731) <223> n equals or c a,t,g, <220>

<221> SITE

<222> (2734) <223> n equals or c a,t,g, <400> 35 ggctcggaag ccccggagtcagattaagaatgagatcaacattgacaccctggccagaga 60 tgaattcaac ctccagaagatgatggtgatggtaacagcctcaggcaagctttttggcat 120 tgagagcagc tctggcaccatcctgtggaaacagtatctacccaatgtcaagccagactc 180 ctcctttaaa ctgatggtccagagaactactgctcatttcccccatcccccacagtgcac 240 cctgctggtg aaggacaaggagtcgggaatgagttctctgtatgtcttcaatcccatttt 300 tgggaagtgg agtcaggtagctcccccagtgctgaagcgccccatcttgcagtccttgct 360

24 tctcccagtcatggatcaagactacgccaaggtgttgctgttgatagatgatgaatacaa 420 ggtcacagcttttccagccactcggaatgtcttgcgacagctacatgagcttgccccttc 480 catcttcttctatttggtggatgcagagcagggacggctgtgtggatatcggcttcgaaa 540 ggatctcaccactgagctgagttgggagctgaccattcccccagaagtacagcggatcgt 600 caaggtgaaggggaaacgcagcagtgagcacgttcattcccagggccgtgtgatggggga 660 ccgcagtgtgctctacaagagcctgaaccccaacctgctggccgtggtgacagagagcac 720 agacgcgcaccatgagcgcacctttattggcatcttcctcattgatggcgtcactgggcg 780 tatcattcactcctctgtgcagaagaaagccaaaggccctgtccatatcgtgcattcaga 840 gaactgggtggtgtaccagtactggaacaccaaggctcggcgcaacgagtttaccgtact 900 ggagctctatgagggcactgagcaatacaacgccaccgccttcagctccctggaccgccc 960 ccagctgccccaggtcctccagcagtcctatatcttcccgtcctccatcagtgccatgga 1020 ggccaccatcaccgaacggggcatcaccagccgacacctgctgattggactaccttctgg 1080 agcaattctttcccttcctaaggctttgctggatccccgccgccccgagatcccaacaga 1140 acaaagcagagaggagaacttaatcccgtattctccagatgtacagatacacgcagagcg 1200 attcatcaactataaccagacagtttctcgaatgcgaggtatctacacagctccctcggg 1260 tctggagtccacttgtttggttgtggcctatggtttggacatttaccaaactcgagtcta 1320 cccatccaagcagtttgacgttctgaaggatgactatgactacgtgttaatcagcagcgt 1380 cctctttggcctggtttttgccaccatgatcactaagagactggcacaggtgaagctcct 1440 gaatcgggcctggcgataaagaacaaagactgtgcctaaaagtggagagccaggggagtg 1500 tgggtcagataagcagctacagctgcagtttggtggattggtggagtatgtgtgtgtgtc 1560 agtgctcagctaagaactgtagggaagatggatgaccttcacgcagaactccttttggga 1620 tatacatgatgcagaaaggatcctacatggagagagacagaactctctcagctgacactc 1680 tcagagattcctgatgggctttctcttgaagtccaaaggcgtctgcattgtttcctttct 1740 ttgcccatccatgaatgttctgttttgttttgttttttttaataagaattccggctgatt 1800 tttgtgaggcctgtttaaattgactttactttgccttttgtgtttctcaattttatctag 1860 aaatctttctgactttttccatctcttgcttcaaagtaagaggggaactctccttgccga 1920 ctccaccttataggtacatttggtgttttgcactgggaagaaataggatccatccttagc 1980 tgaggcttgaggactgatccagcctctcatggcttccctccaaagtaacttagggttgag 2040 ggatctatatgtgatgtcaaaacttactttaaacctctagtttcgtgctgtcatttatta 2100 ggctgggccaccaaatctttgtttcaatttatcagaagccaagtgcatactagcgtcttg 2160 tttgttgcccattgcctatacttttcacctgagatgtgtgagttggggccttttaaaaac 2220 tactgaattgtctgagccttgaagacatttccagggagaagagataatctctcatttcac 2280 ccacaggctggtctaatcataacctagttaaagatgtccttgtttaagaaccccattatt 2340 tatttttagtttttaatataaattaacatgtgggtcattatatttctccttaaatgagga 2400 aattttaaattttattgatctaacctttgaagctttaaaaaaggagaaagagggtagggg 2460 tgggaaactggcatactgtgtgtatagcactgccgattggctaggccactgtgtctctgc 2520 tacaaattaaagaaatcctaaaagttttccttggtcatagagttggggaatgacagaatt 2580 tttctttgttgtgaaatgtatgtacagagtagaccatctctagccctgtggtgaaagagg 2640 tacactcgaatgtttgcataaagcaagtgacaaatgaaaaaaaaaaaaaaaaaaaaaccc 2700 cgggggggggcccggtaacccatttggccnnaanggg 2737 <210> 36 <211> 1507 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (486) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1237) <223> n equals a,t,g, or c <400> 36 ggggtcaaccacgagagctggcgtgacagtggccagactgacggcctgacctttggccac60 ctgaagatggtgctgctgtgggcctctgtgctcttcccggcgcccgaggactgggcagag120 ctgcagggcgccgtgtaccgcctgctggtggtgctgctctgttgcctggccacgcggaag180 ctgccccact.tcctccacccgcagcgcaacctgctgcagggcagcggcctggaccttggt240 gccatctaccagcgcgtggagggcttcgccagccagcccgaggcggccctgcgcatccac300 gccacccacctgggccgcagccccccgccgcgcatcggctccgggctcaaggcgctcctg360 cagctgccagccagtgaccccacttactgggccactgcctacttcgacgtcctgctggac420 aagttccaggtcttyaacatccaggataaggaccggatctctgccatgcagagcatcttc480 cagaanaccaggactctgggaggcgaggagagctgagctgggccacctggtctcagccac540 ctgttcttggctccccaacagactctgcactgcaccatgggaggctcctgggatgtttgg600 aagaagaaacgggcttctccttgagggggtagtggagggattttgtccccagcagtggcc660 tctgagagtctttcagtgcctggtggggcagggcaggcctcttggagcacctcctccctg720 ggtcagggcctggatgcaggtgccaagctctccatgtggtgcatgttgacccagccacgt780 ggtgttgtcaagcaaacagcatcggcaggagacctggagctgaggacttggccctgcctg840 cactgtatgccattccttggtgacraaatgctgtatatttggttttgaaaaaatgaatgt900 gctgggtatacacagcagaaagggtactgtccactttttgtatatcagtgtggaaaatat960 ttcccctagaagtagaaaaggctcatgtgctgatggataattttgagtcttctccattct1020 ctgtgaatgacccccttccccaggcatccccacctcctacctcgttcttagagcaaacta1080 aagccaactgagggtgcacacacagccatgagcccacctgcccaggactactcccatctg1140 cttcttcccgtccccgtggaagtggcccctgatatggattgtatctgtatccccacccaa1200 atctcaggtagaattggatctgtgtccccacccaagntytcaggtagacttgtaatcccc1260 agtgttggaggaggggcccagtggaggtgattggatcatgggggtggatttctcccttac1320 tggtctcatgatagtgagtgagttctcagagatctggktttttgagtgtgtggccatccc1380 gtctcttttgcctcttctctttctcggcatgtagaatgctcatctcttcacttcaccaga1440 tgcaagtccgagctccagcaagttccgatagctcggatggagcaataaccctgaggataa1500 aaaagaa 1507 <210>

<211>

<212>
DNA

<213> sapiens Homo <220>

<221>
SITE

<222>
(651) <223>
n equals a,t,g, or c <220>

<221>
SITE

<222>
(750) <223>
n equals a,t,g, or c <400>

tatttaggtgacactatagaaggtacgcctgcaggtaccggtccggaattcccgggtcga 60 cccacgcgtccggcgggggcatcgccgcccgcgcccctctaagtgccgggccgcaagctc 120 caccgcagccgcctgcaagcagcggcgcctcggccctcgacctgcgcgcaaagcctgtgc 180 tggagccgtcctcccgcggcggggaccgggaccggggacccaagccaatcgaaagctcca 240 accatggccatggggctcttccgcgtgtgtctggtggtggtgacggccatcatcaaccac 300 ccgctgctgttcccgcgggagaacgccacagtccccgagaacgaggaggagatcatccgc 360 aagatgcaggcgcaccaggagaagctgcagctggagcagttgcgcctggaggaggaggtg 420 gctcggctggcggccgaaaaggaggcactggagcaggtggcggaggagggcaggcagcag 480 aacgagacacgcgtggcctgggacctctggagcaccctctgcatgatcctcttcctgatg 540 atcgaggtgtggcggcaggaccaccaggaggggccctcacctgagtgcctgggcggtgag 600 gaggatgagctgcctggctggggggcgcccccttgcagggcctcaccctgnccaacaagg 660 cacgcttgccacttttatgagcgctgcatccggggggccacggccgatgcagcccgtacc 720 cggragttcctggaaggcttcgtggatganttgctggaagccctgaggagcctctgcaac 780 cgggacaccgacatggaggtggaggacttcattggcgtggacagcatgtacsagaactgg 840 caggtggaca ggccactgct gtgccacctt tt 872 <210> 38 <211> 1816 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (1807) <223> n equals a,t,g, or c <400>

ggcggccgctctagaactagtggatcccccgggcaggaattcggcacgagcgtgatcatc 60 gggatgctcgtgctcctgctggactttcttggcttggtgcacctgggccagctgctcatc 120 ttccacatctacctgaaggccaagaagatgaccacctttgagtatctcattaataaccgc 180 aaagaagagagttcaaaacatcaagcagtgaggaaagatccatacgtgcaaatggacaaa 240 ggagttctccagcaaggagctggcgccctgggctcatctgcacagggagtcaaagccaag 300 agctccctgctgattcacaagcacttatgtcacttctgcacttcagtaaaccaggatggg 360 gattcgacggcacgggtgcatttgtgagtcccctcagtgtctctgcagcttctatgtgtg 420 gatgaatagtgaagccacatgaggcctggtctgaagcagagaagatccgctcagcatcat 480 gttgaatcccaagcccgcgtctccgtgggctccaggacaatcctatgaaagtgacaccgt 540 cgttcattgttcacatcggggaggagaattccatctgaaaatgcacgtgacttcactgac 600 acccgcgtccgtggcacagccctgtgctgagctccacagacctacagtccatcgcctccc 660 cttggagtgggcccacagctgcagacagtgttgtccaaagaagatgctctgagtagatga 720 atggaacaaactggaaactccagaaacaagcccttctttacattyctaatgtttggctta 780 aatggaaggtgccaagagccaaggccaggaactcccggctgatttgcaggcgcctgtgtc 840 agttctccactcgtgtacacccagacgggggctcgatggcacaggaagcagatgatgccc 900 cgagtatatctacacttgggctgcaacaagaaacaacagagcccatgaaaactgacagtg 960 ctgaaagtgaagactgagattcaggagctsaggtgcccctgtgatccaggtcttcyaccc 1020 tgaaaccccaycctccatcaaggtcctgcctgtagagtctaccttgcaaagcctcctgct 1080 cctacccatgctacaggccaggaaccagagcccatcatctcagaggcccctggatgtcct 1140 tcgaaggaaccaggaccctcagagcccagcatccatctctgtcatcatcttcatcacacc 1200 caaagaagagccagccttgcaggagggtttacatctccaggaagatgggctgccagcaac 1260 tgcagaggatgcagccacctgcttaactgtgctgtccagccagccagccagctgcagggc 1320 ctcttgctgcttaagagctgatgggccgggcatgttggctcacacctgtgagcacagtac 1380 tgggaaatgggagcacagtactaggaaatgggagcacagtactgggaaatgggagcacag 1440 tactgggaaatgggggctcacagcactgcaaaatgggagcacagtattgggaaatgggag 1500 cacagtactgggaagtgggagcacagtactgagaagtgggagcacagtactgagaaatgg 1560 gagtacactactgagaaatgggagcacagtactgggaaatgggcatacagtactggggaa 1620 atgggcatacagtactgaraaatgggarcacagtactgggaaatgggagcacagtactgg 1680 ggaaatgggarcccacagtwctgggaaaagggartttcacattactccggaaattgggar 1740 cmtaktactggggaawtgggagcacrttactggggaaatgggaaacattattactgggga 1800 aaccccngaacctgga 1816 <210> 39 <211> 631 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (337) ' <223> n equals a,t,g, or c <220>

<221> SITE
<222> (338) <223> n equals a,t,g, or c <400> 39 gggaaatgggagctcacagcactgggaaatgggagcayagcaytgggaaatgggagcaca 60 gtactgggaattgggagcacagtactgggaaatgggagctcacagtactgggaaatggga 120 gctcacagtactgggaaatgggagcacagtattcggaaatgggagcacagtactgggaaa 180 tgggagctcacagcactgggaaatggagcatagcattgggaaatgggagcacagtactgg 240 gaaatggaagctcacagtcctgrgaaatgggagcayagtrctgggaaatgggagcacagt 300 actgrgaaatgggagcacagtactgggaaakgggagnncmcagtactgmggaaatgggag 360 cayagtactgggaaatgggagcacagtactgggaaatgggagcatagtactgggaaaccc 420 cagacctggattctgagtttttcagcctagcccagacttcttatcttagtagacaaaaag 480 agtcaataccagagaaccagaggcatcctctgtattttaatgaactctgcattttaatct 540 gtttagtagtcattttttaaaagataatcagttttccaaatatatctataagttactacg 600 tgcaaaaaaaaaaaaaaaaaaaaaaactcga 631 <210>

<211>

<212>
DNA

<213> sapiens Homo <220>

<221>
SITE

<222>
(2) <223> als a,t,g,or c n equ <220>

<221>
SITE

<222>
(31) <223> als a,t,g,or c n equ <220>

<221>
SITE

<222>
(87) <223> als a,t,g,or c n equ <220>

<221>
SITE

<222>
(92) <223> als a,t,g,or c n equ <400>

cnaaacgcttcatttttaggcgtttgggtcngttctccactcgtgtacacccagacgggg 60 gctcgatggcacaggaagcagatgatnccccntgtatatctacacttgggctgcaacaag 120 aaacaacagagcccatgaaaactgacagtgctgaaagtgaagactgagattcaggagctc 180 aggtgcccctgtgatccaggtcttctaccctgaaaccccaccctccatcaaggtcctgcc 240 tgtagagtctaccttgcaaagcctcctgctcctacccatgctacaggccaggaaccagag 300 cccatcatctcagaggcccctggatgtccttcgaaggaaccaggaccctcagagcccagc 360 atccatctctgtcatcatcttcatcacacccaaagaagagccagccttgcaggagggttt 420 acatctccaggaagatgggctgccagcaactgcagaggatgcagccacctgcttaactgt 480 gctgtccagccagccagccagctgcagggcctcttgctgcttaagagctgatgggccggg 540 catgttggctcacacctgtgagcacagtactgggaaatgggagcacagtactaggaaatg 600 ggagcacagtactgggaaatgggagcacagtactgggaaatgggggctcacaggactgca 660 aaaatgggagccagtattgggaaaatgggaagcacagtacttggggaagtggggagcaca 720 gtactgagaagtgggagcacagwactgagaaatgggagtacactactgagaaatgggagc 780 acagtactgggaaatgggcatacagtactgggaaatgggcatacagtactgagaaatggg 840 agcacagtactgggaaatgggagcacagtactgggaaatgggagcccacagtactgggaa 900 aggggagttcacagtactcggaaatgggagcatagtactgggaaatgggagcacagtact 960 gggaaatgggagcatagtactgggaaaccccagacctggattctgagtttttcagcctag 1020 cccagacttcttatcttagtagacaaaaagagtcaataccagagaaccagaggcatcctc 1080 tgtattttaatgaactctgcattttaatctgtttagtagtcattttttaaaagataatca 1140 gttttccaaatatatctataagttactacgtgaaaaaaaaaaaaaa 1186 <210> 41 <211> 1516 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (7) <223> n equals a,t,g, or c <400> 41 ttgggcnccaaaaatcaacgggacttttccaaaatgtcgtaccaactccgccccattgac60 gcaaatggccggtaggcgtgtacggtggaaggtctatataagcagagctcgtttagtgaa120 ccgtcagatcgcctggagacgccatccacgctgttttgacctccatagaagacaccggga180 ccgatccagcctccggactctagcctaggccgcgggacggataacaatttcacacaggaa240 acagctatgaccactagscttttgcaaaaagctatttaggtgacactatagaaggtacgc300 ctgcaggtaccggtccggaattcccgggtcgacccacgcgtccggagcgaggctgccctt360 tcttcgcagcgtgatttattttcttctttttttctgaactcttcttccagggagaggcta420 gtggtaacaggccgagctggatggatgggtatggggagaggggcaggacgttcagccctg480 ggattctggccgaccctcgccttccttctctgcagcttccccgcagccacctccccgtgc540 aagatcctcaagtgcaactctgagttctggagcgccacgtcgggcagccacgccccagcc600 tcagacgacacccccgagttctgtgcagccttgcgcagctacgccctgtgcacgcggcgg660 acggcccgcacctgccggggtgacctggcctaccactcggccgtccatggcatagaggac720 ctcatgagccagcacaactgctccaaggatggccccacctcgcagccacgcctgcgcacg780 ctcccaccggccggagacagccaggagcgctcggacagccccgagatctgccattacgag840 aagagctttcacaagcactcggscacccccaactacacgcactgtggcctcttcggggac900 ccacacctcaggactttcaccgaccgcttycagacctgcaaggtgcagggcgcctggccg960 ctcatcgacaataattacctgaacgtgcaggtcaccaacacgcctgtgctgcccggctca1020 gcggccactgccaccagcaagctcaccatcatcttcaagaacttccaggagtgtgtggac.1080 cagaaggtgtaccaggctgagatggacgagctcccggccgccttcgtggatggctctaag1140 aacggtggggacaagcacggggccaacagcctgaagatcactgagaaggtgtcaggccag1200 cacgtggagatccaggccaagtacatcggcaccaccatcgtggtgcgccaggtgggccgc1260 tacctgacctttgccgtccgcatgccagaggaagtggtcaatgctgtggaggactgggac1320 agccagggtctctacctctgcctgcggggctgccccctcaaccagcagatcgacttycag1380 gccttccacaccaatgctgarggcaccggtgcccgcargctggcagccgscagcctggac1440 ccacagsccccgagamcttyccatacsagacaggccgtggccaagtgcaaggagaagctg1500 ccggtggaggacctgt 1516 <210> 42 <211> 803 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (462) <223> n equals a,t,g, or c <220>

<221> SITE

<222> (696) <223> n equals a,t,g,or c <220>

<221> SITE

<222> (746) <223> n equals a,t,g,or c <220>

<221> SITE

<222> (761) <223> n equals a,t,g,or c <220>

<221> SITE

<222> (770) <223> n equals a,t,g,or c <220>

<221> SITE

<222> (780) <223> n equals a,t,g,or c <220>

<221> SITE

<222> (788) <223> n equals a,t,g,or c <400> 42 gactaccttt tgcawaagctatttaggtgacactatagaa ggtacgcctg caggtaccgg60 tccggaattc ccgggtcgacccacgcgtccggagcgaggc tgccctttct tcgcagcgtg120 atttattttc ttctttttttctgaactcttcttccaggga gaggctagtg gtaacaggcc180 gagctggatg gatgggtatggggagaggggcaggacgttc agccctggga ttctggccga240 ccctcgcctt ccttctctgcagcttccccgcagccacctc cccgtgcaag atcctcaagt300 gcaactctga gttctggagcgccacgtcgggcagccacgc cccagcctca gacgacaccc360 ccgagttctg tgcagccttgcgcagctacgccctgtgcac gcggcggacg gcccgcacct420 gccggggtga cctggcctaccactcggccgtccatggcat anaggacctc atgagccagc480 acaactgctc caaggatggccccacctcgcagccacgcct gcgcacgctc ccaccggcgg540 agacagccag gagcgctcggacagccccgagatctgccat tacgagaaga gctttcacaa600 gcactcggsc acccccaactacacgcactgtggcctcttc ggggacccac accttcagga660 ctttcaccga ccgctttcagacctgcaaggtgcaanggcg cctggccgct catcgacaat720 taattacctg aacgtgcaggtcaccnacacgcctgtgctt nccggctcan cggccactgn780 caaccaanca aggcttaccatca 803 <210> 43 <211> 1553 <212> DNA

<213> Homo Sapiens <220>
<221> SITE
<222> (23) <223> n equals a,t,g, or c <220>
<221> SITE

<222> (41) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (42) <223> n equals a,t,.g, or c <400>

tgcacgcgtaagcttgggccccncgagggatcctctagagnngtcgcscttttttttttt 60 ttttttaaaagaaaataacaaaaaaaccaaactttacttgcatttagccattaataaata 120 atttacagtatgtacacgcggtgacactccacacaggtcgcagacacgtggacgcagtga 180 ggggtccccacttccaagcagaacgtgagcaaacacaaccaaaataaagtgcttcacttt 240 ttacttccaacatagggaccaactaaaaccagcagggagagggaggggccgggccccgag 300 cgccctcccacccgcccacacacggttctggcagagggagcaggacctcttggagggagg 360 gggagacacacgttctgcagcctcctggccagtgtatgagaggtccctggggaacagcag 420 aggggctgccgatgggtgggtagggtgtggagctgtagggctgctggcaggatcaccctc 480 cagggcctggctgcacttttgggagtgaagcctggctttgtgctgtgggcatagcagcac 540 aggtgggactgttctgggcccctggatgagcaccctgggctgaaatgaggcgggagccag 600 cggcaagtctctcctggctgtgtggccacagggcttaagggaagagattagaggttgaag 660 accctgccagggatgctcagagctgagagtcctaagaccctgctcctctccccagactcc 720 agacgtggatggaccgggctgggggctcttggggttgtgaggaggaagcagaggatcagg 780 gcaggccagctggccccatccactgggtctcctgccacaccctggggcccaggcacccct 840 gcccaggagctcttgcatctgcaggagggagaggaacagaagtcaaggatctcttctctg 900 tttagacatttagaaccaagcacttcgactttctaagcacagggtggaaggggcaggggc 960 ccaggccagagacgggcagctgagcaggtgcctgaaaaccgggacctgcactggtaccca 1020 cagcagccaggtgaacatgggcagggcagccacaggccagaaggcaccaggcgggcacga 1080 cctactggccaaaaggttccagcagtctgctaaggtgccaagcagcgccccggggttgcc 1140 ctgcctsgggccaagccccaaggatgcctccctgtgtggctggggtgtccccgctggcct 1200 agggggatcgagtgtgctctcgtgtaagagtgtgtgcgtgcgtgtggcgggcacaggggg 1260 ccccacggatcggcgagcagcagtcggccgggcctttcagtgcattgcgagggggaagga 1320 gctgactctgacggttcccagtgtgtctctggtgggggtggggggcttcagcctgagggg 1380 atgtttcacacattcacactgcaggggcggggcgaggggagctgctctccctctcacaca 1440 gctctactgtcaaacatcacggcaccagtcaccacaaccttgtcacgtgcactagaaggg 1500 gaggggtccatgcctgagcccttggcagcaggcggtccctggcgttctgcggg 1553 <210> 44 <211> 368 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (365) <223> n equals a,t,g, or c <400>

ggagaacaggggcaatgacacatgctccttggcttcctagtcctgatcccttggggttcc 60 ttgatcctagggtcatctgacttagatccatcctcccttcctctaggcaccaggggccat 120 ggctggaggtggccacccctcagtcctgtacagattctgtacccactggctggagacccc 180 catgcagctgtctcctgctcttgctgcggggagactgaactcagggctctgctcactggc 240 tccctcccaatggaagccttctctgggcttcacagcattgaatattcctccagaacagct 300 tgttaaatcaaattttttttttttttagatggagycttgctytgkcgcccaggctggaat 360 gcacnggc 368 <210> 45 <211> 1447 <212> DNA

<213> Homoapiens S

<220>

<221> SITE

<222> (4) <223> n ls a,t,g, equa or c <220>

<221> SITE

<222> (887) <223> n ls a,t,g, equa or c <220>

<221> SITE

<222> (903) <223> n ls a,t,g, equa or c <220>

<221> SITE

<222> (1033) <223> n ls a,t,g, equa or c <220>

<221> SITE

<222> (1137) <223> n ls a,t,g, equa or c <220>

<221> SITE

<222> (1371) <223> n equals a-,t,g, or c <220>

<221> SITE

<222> (1413) <223> n ls a,t,g, equa or c <400> 45 tccnttgggaataaccctcactaaagggaacaaaagctggagctcgcgcgcctgcaggtc 60 gacactagtggatccaaagaattttggcacgagcacaaactgggtggcttaaaccacaga 120 aatgtattgtcttacagttctgggggccagaagtccaaaatcaaggtgtcagcaggccca 180 tgctccatttgaaggcactggggaggggtctgctccaggcctgtctcctggctactggtg 240 gctccatggcttgtgacagcatcactccaatcttcacacagcattctccccgtgtgcctc 300 tgagccccagttttcccttacttcaagagactggggcttttgctccatcacctagactgg 360 agggcagtggcatgatcttggctcactgctgccttgaactcctgtgctcaagtgatcctc 420 ctgcctcggccatccagaattgctgggactatagatgtgagccacgacacctggcttcaa 480 tttcctctttctataaggatatcggtcgtattggattagggcccgtcctactccagtata 540 acctcatcttagygaattacatctgccacaactctgtttccaaataaactcacattctga 600 ggtacagggagttaggacctcaacacaggaattttgggggctaaggtaaggagaaaacaa 660 gaagagtttggtgtcacagaagtgagaatgtccggccaggtatggtggctcaagcctgtg 720 atcctagcactttgggaggccgaggtgggcagatcacctgaggtgagcagttcaagacca 780 gcctggccaacatggtgaaactctgtctctactaaaaatacaaaaattagctgggccgtg 840 gtggcagacacctataatcccagctatccgggaagctgaggcagganaatcacttgaacc 900 cangagatggaggctgcagttgcagtgagccaagatcacaccactgcactccagcttggg 960 tgacagagggagatgccgtctcaaaaaaaaaaaaaaaaaaaaaaaaactcgagagtactt 1020 ctagagcggccgngggcccatcgattttccacccgggtggggtaccaggtaagtgtaccc 1080 aattcgccctatagtgagtcgtattacaattcactggccgtcgttttacaacgtcgngac 1140 tgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagc 1200 tggcgtaatagcgaaaaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaat 1260 ggcgaatggagatccaatttttaagtgtataatgtgttaaactactgattctaattgttt 1320 gtgtattttagattcacagtcccaaggctcatttcaggcccctcagtcctnacagtctgt 1380 tcatgatcataatcagccataccacatttgtanaggttttacttgctttaaaaaaccttc 1440 cacacct 1447 <210> 46 <211> 425 <212> DNA

<213> Homo Sapiens <220>

<221> SITE

<222> (352) <223> n equals a,t,g,or c <220>

<221> SITE

<222> (362) <223> n equals a,t,g,or c <220>

<221> SITE

<222> (369) <223> n equals a,t,g,or c <220>

<221> SITE

<222> (415) <223> n equals a,t,g,or c <400> 46 ggcacgagca caaactgggtggcttaaaccacagaaatgtattgtcttac agttctgggg60 gccagaagtc caaaatcaaggtgtcagcaggcccatgctccatttgaagg cactggggag120 gggtctgctc caggcctgtctcctggctactggtggctccatggcttgtg acagcatcac180 tccaatcttc acacagcattctccccgtgtgcctctgagccccagttttc ccttacttca240 agagactggg gcttttgctccatcacctagactggagggcagtggcatga tcttggytca300 ctgctgcctt gaactcctgtgctcaagtgatcctcctgcctcggccatcc anaattgctg360 gnactatana tgatgagccacgacacccttggcttcaatttcctctttct ataangatat420 cgctc 425 <210> 47 <211> 1530 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (1389) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1488) <223> n equals a,t,g, or c <400> 47 ggtgttggcttcctcatgcattctcaatgccaaggctttttctcttctctaacgatgctg60 aatcctctgagacagctctttaagctgatggcctctctgtttctgtcggtgttcacactt120 ggccttccctttgccctctttcagtattatgcctacacccaattctgtctgccaggctca180 gcccgccccattcctgagcctttggtacagttagctgtagacaagggctaccggattgca240 gagggaaatgaaccgcttggtgcttctgggatgttccactaatatacagctatawcyagg300 atgtctactggaatgttggctttttgaaatactatgagctcaagcaggtgcccaattttc360 tactggctgcaccagtggctatactggttgcctgggcaacttggacatacgtgaccactc420 acccttggctctgccttacacttgggctgcaaaggagcaagaacaataagaccctagaga480 agcccgatcttggattcctcagtcctcaggtgtttgtgtacgtggtccacgctgcagtgc540 tgctgctgtttggaggtctgtgcatgcatgttcaggttctcaccaggtttttgggctcct600 ccactcctattatgtactggtttccagctcacttgcttcaggatcaagagccgctgttga660 gatccttaaagactgtgccttggaagcctcttgcagaggactccccaccaggacaaaagg720 tccccagaaatcctatcatgggacttttgtatcactggaaaacctgttctccagtcacac780 gatacattctaggctacttcctgacttactggctcctgggactactcctacattgcaact840 tcctgccttggacatgacctggactctccagggacaggttggaagccaacttaacccagg900 ggtctgaaagtaaaaatacacattggaactgcctctgctgccctgggatcattactgtgt960 ccattataatctttctctttctctttgaaagctggtcaggaatgggagaagtgtcagaca1020 ctagagagccccttctggtcctggctagggcaaattttagacaactattttctctgtaag1080 tgaagattgtcgtattccaagtctaaaatacacctggatctgtctagtcaatcaacatag1140 cagagacagtcttaaacctaccattgacctgtgtgtaaatttaaatgtcaatttattgaa1200 gtgtaaatttcatcaaaggcattagctgacaggctggtaacagtccacacaagatggtat1260 aggcctgaacagtgtagtggcagtaataaagtgggaccattttttccaaatgcgcatgtt1320 ctgatgtttttatgatgtgaggatttattgaaaactcaagttcgatactctcagtacatg1380 aaaaaaaana.agaagaaaaaaaaaaagtttttgccgggtcctttggttcttcctgggtct1440 gggctcgcctagggacctcgggatggccgcctatcgagggctcgcggnggcggtccggtg1500 tttgcccagggtccaaggtctgtccgcttg 1530 <210> 48 <211> 100 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (80) <223> n equals a,t,g, or c <400> 48 gccaccgcgg tggcggccgc tctagaacta gtggatcccc cgggctgcag gaattcggca 60 cgaggtgttg gcttcctcan gcattctcaa tgccaaggct 100 <210> 49 <211> 550 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (18) <223> n equals a,t,g, or c <220>
<221> SITE

<222> (528) <223> n equals a,t,g, or c <400> 49 gggaacgcttttacgccngcaggtcccggtccggaattcccgggtcgacccacgcgtccg 60 gaggtgkkgggcagagtccccakkcagttgggattccggggagggaccacagtgctggga 120 accacagcactgacgccatacacaccaggggatgtttgctcttgcatggaaagtgatttt 180 cagtgtcatgctgcagaatcccatacgttatccttcagtccttggtataaagtcctccct 240 gctcagtagtcttgtgttggtgatggtttggggaaatgaaaagagtgggccctgccccac 300 accaaagtccaggaagggaagaaggagctgtccagcacaggtggggaggggagaaraggg 360 gtcctattgggacccagaattcagactgagcaggaaatccaaccaggggttgaggaggga' 420 ttacctgagcttgtatcacttcaacctgcattttagagacacattctaatctcacagatg 480 cagaatttggggatccctggacagctaacacttgggggtgcaaatgtncttgttgaatga 540 gaggaactgg 550 <210> 50 <211> 866 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (833) <223> n equals a,t,g, or c <400> 50 gaatacgccagctctaatacgactcactatagggaaagctggtacgcctgcaggtaccgg 60 tccggaattcccgggctcttgatctttcctggtcctggtacggggatgacctctctttta 120 gaggggcgaatggttctctgtgtctcctgcctcctccttcccctgctgttgttgctgaaa 180 cactttaatgggttgatgacaccatatttagctcataatgtatattgtcctatagaatat 240 atttcttttttcccatttcatgaaaaaaatatagaatatatttctatttggttcatcttt 300 gacagttttaagtttatctattcgaggcttttgtgtatttcccaaatttatgtgctttat 360 agagcatatactttgccccattagatgcagtggtattgcaatagttatcctcagggaggt 420 aggctgtgtgtgtgtgtgggtgtgagtgtatgtatttgtataaaagaaaatccaatatta 480 catgcatttggccgggcacggtggctcacatttgtaatctcagtgctttgggaggctgag 540 gtggacaggtcatttgaggtcaggagttcgagaccagcctggccaacatggtgaaactct 600 gtctctactaaaaattaaaaaattagccaggcgtggtggtgtacatc.tgtaatcccagct 660 acctgggaggctgaggcaggagaatctcttgaacctgggaggtggaagctgcagtgagcc 720 aagattgcaccactgcactccaggctgggggacagagcaagactccggtctcttcaaaaa 780 aaaaaaaaaaaaaamaaamaaraaaaaaagggcggccgctctagaggatccangcttacg 840 tacgcgtgcatgcgacgtcatactct 866 <210> 51 <211> 732 <212> DNA
<213> Homo Sapiens <400> 51 cttgatctttcctggtcctggtacggggatgacctctcttttagaggggcgaatggttct 60 ctgtgtctcctgcctcctccttcccctgctgttgttgctgaaacactttaatgggttgat 120 gacaccatatttagctcataatgtatattgtcctatagaatatatttcttttttcccatt 180 tcatgaaaaaaatatagaatatatttctatttggttcatctttgacagttttaagtttat 240 ctattcgaggcttttgtgtatttcccaaatttatgtgctttatagagcatatactttgcc 300 ccattagatgcagtggtattgcaatagttatcctcagggaggtaggctgtgtgtgtgtgt 360 gggtgtgagtgtatgtatttgtataaaagaaaatccaatattacatgcatttggccgggc 420 acggtggctcacatttgtaatctcagtgctttgggaggctgaggtggacaggtcatttga 480 ggtcaggagttcgagaccagcctggccaacatggtgaaactctgtctcta ctaaaaatta540 aaaaattagccaggcgtggtggtgtacatctgtaatcccagctacctggg aggctgaggc600 aggagaatctcttgaacctgggaggtggaagctgcagtgagccaagattg caccactgca660 ctccaggctgggggacagagcaagactccggtctcttcaaaaaaaaaaaa aaaaaaaaaa720 aaaaaaaaaaas 732 <210>

<211>

<212>
DNA

<213> Sapiens Homo <220>

<221>
SITE

<222>
(584) <223> als a,t,g,or c n equ <220>

<221>
SITE

<222>
(595) <223> als a,t,g,or c n equ <400>

gaattcggcacgaggcggcgcgggacctgcagtcgccagggattccctccaggtgacgat 60 gctctggttctccggcgtcggggctctggctgagcgttactgccgccgctcgcctgggat 120 tacgtgctgcgtcttgctgctactcaattgctcgggggtccccatgtctctggcttcctc 180 cttcttgacaggttctgttgcaaaatgtgaaaatgaaggtgaagtcctccagattccatt 240 tatcacagacaacccttgcataatgtgtgtctgcttgaacaaggaagtgacatgtaagag 300 agagaagtgccccgtgctgtcccgagactgtgccctggccatcaagcagaggggagcctg 360 ttgtgaacaktgcaaaggttgcacctatgaaggaaatacctataacagctccttcaaatg 420 gcagagcccggctgagccttgtgttctacgccagtgccaggarggtgttgtcacagagtc 480 tggggtgcgctgtgttgktcattgtaaaaamcctttggagcatctgggaatgtgctgccc 540 cacatgtccaggctgtgtgtttgagggtgtgcagtatcaagaangggaggaattncagcc 600 agaa 604 <210> 53 <211> 854 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (805) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (818) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (829) <223> n equals a,t,g, or c <220>
<221> SITE

<222> (832) <223> n equals a,t,g, or c <400> 53 tcgagttttttttttttttttttttttttgcttttacatcttaagccctttattgactac 60 aatgcagaacattttattttaagacacagtgggttttgtttttgttgatgttttcaccaa 120 ttcaactgaagacgaaagtcaagacaatcaaatggtaactagtagcagcctatcagtaaa 180 tgagggcaagtatagagactgttctttggactgaggttaaatcaattagtcaataaaggc 240 ttttccactgtctaataattataacatattaacagtcgccaaatagtgttggatgggact 300 cctctagaaataactaaagcctttcattttatacatgaaatagccacaaaatgtagatgg 360 gttacatcaactcattggatttgcccatcttaaattactctgagattcagagaaatcaaa 420 atttcatcatatgttgtgcaacagtttcttcaaatactttctcatgctcaataaaattaa 480 catggcatccagacaccctttaatttcaacacatgacatataaatggtcacatccatcat 540 acacacatagatgattagaagacttgaggactacgcaaggatggcaagaaatcacttgat 600 tcttcggttatgccctagtgacaaatatatatttcttggcaaccagccaatcaattttgg 660 gaataatgttagcccatgacctaatacgttctaacagcctatcactcgctttttcatgca 720 ccatttagggccgggtcaccaatctcctgctgttcttttcccaggcatacattcctttct 780 ttgggaaacctggaaatgccccccnggataaatggggncccttctcccnggnggaaccac 840 cattttccccactc 854 <210>

<211>

<212>
DNA

<213> Sapiens Homo <220>

<221>
SITE

<222>
(341) <223> als a,t,g,or c n equ <220>

<221>
SITE

<222>
(354) <223> als a,t,g,or c n equ <220>

<221>
SITE

<222>
(804) <223> als a,t,g,or c n equ <400>

aaggggaggaatttcagccagaaggaagcaaatgtaccaagtgttcctgcactggaggca 60 ggacacaatgtgtgagagaagtctgtcccattctctcctgtccccagcaccttagtcaca 120 tacccccaggacagtgctgccccaaatgtttgggtcagaggaaagtgtttgacctccctt 180 ttgggagctgcctctttcgaagtgatgtttatgacaatggatcctcatttctgtacgata 240 actgcacagcttgtacctgcagggactctactgtggtttgcaagaggaagtgctcccacc 300 ctggtggctgtgaccaaggccaggagggctgttgtgaarantgcctcctacgantgcccc 360 cagaagacatcaaagtatgcaaatttggcaacaagattttccaggatggagagatgtggt 420 cctctatcaattgtaccatctgtgcttgtgtgaaaggcaggacggagtgtcscaataagc 480 agtgcattcccatcagtagctgcccacagggcaaaattctcaacagaaaaggatgctgtc 540 ctatttgcactgaaaagcccggcgtttgcacggtgtttggagatccccactacaacactt 600 ttgacggtcggacatttaactttcaggggacgtgtcagtacgttttgacaaaagactgct 660 cctcccctgcctcgcccttccaggtgctggtgaagaacgacgcccgccggacacgctcct 720 tctcgtggaccaagtcggtggarctggtgctgggcgaracagggtcagcctgcagcagca 780 cctcaccgtgcgctggtacggctngcgcatcgct 814 <210> 55 <211> 322 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (314) <223> n equals a,t,g, or c <400> 55 ggaggaatgaaagtggcggtctcgccggcagttggtcccgggccctggggctcgggagtc 60 gggggcggtgggacagtgcggctactcttgatcctctccggctgcttggtctacggcaca 120 gctgaaactgatgtaaatgtggtcatgcttcaggaatcccaagtttgtgaaaagcgtgcc 180 agccaacaattctgttacacaaatgtgcttatcccaaaatggcatgatatatggacacgg 240 atacagmtccgartamatagttccagattggttcgagtcacccaggtggarraatgarga 300 rgaaactgaaggancttagaac 322 <210> 56 <211> 3140 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (3) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (5) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (21) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1628) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1642) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (1656) <223> n equals a,t,g, or c <220>
<221> SITE
<222> (2094) <223> n equals a,t,g, or c <400> 56 tgntncgagtcacccaggtgnagaatgaggagaaactgaaggagctagagcagatttgat 60 cccaaactctttctkgttttccttcttggacttatgctatttttttgtggagacttgctg 120 agcagaagtcaaattttctactactctactgggatgactgtgggaattgtggcctctctg 180 ctaatcatcatttttatactatctaagtttatgcctaagaaaagtcccatttacgtcatc 240 ctggtgggaggctggtctttttctctgtacctcattcaactagtttttaaaaatttacaa 300 gagatctggaggtgttactggcagtatcttttaagttatgtcctcacagttggattcatg 360 agttttgcagtatgttacaagtatgggcccttggagaatgaacgaagtatcaacctgctg 420 acctggaccttgcagctgatgggcctgtgtttcatgtattctggcatccagataccacat 480 attgcccttgccattatcatcattgctctttgtactaagaacctggaacaccctattcag 540 tggctgtacatcacctgcagaaaggtgtgtaagggagcagaaaagcctgttccccctcgt 600 ctcctgacagaagaagaatatcggatacaaggagaggtagaaacccgaaaggctttagag 660 gagctccgagaattttgtaacagtccagactgctctgcttggaagactgtttctcgaatc 720 cagtctccaaaaagatttgctgactttgtggaaggctcttcccacctcacgccaaatgaa 780 gtttctgtccatgagcaggagtatggattagggagcattattgcccaggatgaaatctat 840 gaggaagcatcctctgaggaggaggactcatattctcggtgtcctgctatcacacagaac 900 aactttctaacctaggtagtggtcagttatctttacgtggactggcttggtgccttggtc 960 catgttgcatgtgttgtgcaattgctttcaaccctttgaaacagagtgagatagataggg 1020 tagaaattctcctactgaaataagaggcctaaaaaggcctccctttggaaatgggaggtc 1080 tctatgggatccctgaggaaggagagtggataaagtagtgaatgctgggtagttcacttc 1140 ccattggttaagctaacagcccacttttatgtttccagagaaattggatggccacagcta 1200 gcatggcattctagctccttcttgaaagttgattcaatcatggcatttctgtcactggct 1260 ggctctccaaagtaagaactgttgttaagtgcaggaatgcttttagactataggctgcaa 1320 cttccagagagaaatccacaaatctgagcctccttcactccagcttttatttcrgtgact 1380 ttagaataattattgatttaactgttttgggaggaaaatagatttttattgttttgtttt 1440 ttaaatgaatgtcttttaaaaamcwtamcaaactcatgttccagaaccagcaagtgctcc 1500 agagtgacacaccccctaggcccctacatatttattaatatggattatccattaaagccc 1560 caggagctgttgttttaagctttgatttagttctcatacatatgatagaaagtcctattt 1620 gcctttangaacatgcctgtangctcttctgcaggntgagatgttctgggctttttatta 1680 tattcaactttcaattccatcttaaaaaacatttgtwttcttctcttcccattcttcctt 1740 accctgcctttgccctttcaggaagggtcagttcccttacctgtgaactatgtatgttca 1800 gagtagcattattcctgctagctaggagaagtcatcttgtttaggggatttggatgcttt 1860 ttatacgttctccattttcctgtcattgggtcatgttatctttgagttgctatgaaatca 1920 ggaaactgtctccttttcctttcccttcctttgtctacatgctctgtccattcctttcag 1980 ccttttctcaccacccatactcccccaaatctgggtaatttttaagccttgaaactatgt 2040 agtttcttgatacacaatttgtagttatgcagcagccacaatttgcattgccanggaaat 2100 argctccaggttatcttcatgcctctgggtgctcattcagctgtcaagtttccatgaact 2160 tacacttatttatgattgcgtttctgacctgagatgtatgctgcctgttattgcagtagc 2220 attagtttcagattcttttgccattgcaaagtaccccttataaaccagcaatgtcatctg 2280 tgaggaagcaaattctcaagtgtctgtcatttacttggttctttttctttgtggtcttca 2340 cccttataccctggaaaagtctgtaattaccttagccaggaagatagatggtcatggcaa 2400 gcgcacagcaccagacttactggctcaccaagatgatggaaaaaggcagatgatttttta 2460 aaaagccgtaatgactcctttagaccagccatttagcgtggtaattttgaaaggcctagc 2520 tccattgcagacttccaaagggtcagctctgagactgccctccaggtgggcagttgatta 2580 tttccaccagtgttttccagagccttaaactgtcctaagtgacaactacctcagttggca 2640 ggaaagagacatatagtagaaagtgaaaaatgagcagtatttgggcagatgctatgggtt 2700 acagttgaagggtaaaaggaactttacattgggaaacctttatacccttgtgaattatgt 2760 acatggtaaaatgttctctctctacaaagaactattaaaacttctgaaatatactatttt 2820 ttaccttatttatagaaattgagacctagcatatttaagcataagtttattttaaaaaat 2880 aattcaactcgtgcaagtggtctcaggattctctggagattttggtgcctcccctactta 2940 gggaggtgatagcttgcctawaagggtgacttttccygatcatgtctttatttcaatgag 3000 aaagcactgtgaaattgtgaaagawtctcctctttctctgtttaataaacccccatgaaa 3060 tataaaaaaaaaaaaaaagggcggccgctctagaggatccctcgaggggcccaagcttac 3120 gcgtgcatgcgacgtcatag 3140 <210> 57 <211> 1402 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (1335) <223> n equals a,t,g, or c <400> 57 ccgggccccccctcgagggtcatttttttttttttttttttttaacctttcaaaatattt60 ttatcattgtgaattggtaattgtattacagtcagtatcctttggggaaaaaagtctcaa120 aatttaacttcccttgttgaagaagctattttacagtgccatattattcattaagcatta180 attaaagaacagcaggataattagtaggatcatcaatattacaagaaacattagatcgct240 caagaaggaaataatttaactagaaacactttaaattttctcaagtagtgcagctgatag300 attagggagaaaaacaacaactccgcttaggggatatagactttaaatcattgtttagaa360 ataggtacttctgtatttattttgttaagttaaaataatagggctcaattaggctgagta420 actccctagaggcacagttgtaatagctggtctcagagaaggcaaaccctcctagttctc480 atcagaaggaaatgatctgtcattagtcctaggtaactgattaatatctcacagtggact540 ggaaatcgctgaatgttctctgcaagttttatactcaaggtgtgaattcataagcaacca600 tcctatttgaaagttagccattgcttagggttctctattagaatcttatcaagcacattc660 tcagttatgcctctcagcaacattttaggaacaacattggtgagtatatgagaatagccg720 tgacctccatatttcatcagccgggttttcgtatgtatgtcatgtgctaccagaattcga780 tcttcacagccctcttccaccaggagacgcacccttctaattcttttgttatcatcaggc840 atgtcaatatctgggccgagttggtaatgaagtagttcagtaccaaagagatcatattcc900 aagtagcagccaagttgagcaaactccaagagctctttcttatcaagaatagtcctatcc960 aggtgtgacatgactgttttggagatgtctgcgcctgcttcttgcaatattcggataatc1020 tgaaatggtgccctggagctccgtccaggatggataataacaggacaaccaagctgagcc1080 tgggcatgagctgtggcctggagaacctttctttcactctcagtcaaaggccaggagcaa1140 ccaatttctccaataatgccacacttgatactggttccatcagctccatggagaatttca1200 ttcataaggacatcggtaagctcgggtacaacgcgctgccttctccctgggaacttccgg1260 caggccaacttccctggcctttttagtctcccctgtgccctcccacttcgccctctggtt1320 ctcctcgtgccgaantcctgcagcccgggggatccactagttctagagcggccgccaccg1380 cggtggagctccagctttggas 1402 <210> 58 <211> 2306 <212> DNA
<213> Homo Sapiens <400> 58 ggcacgagactgcatgcacctgggccttgtcagtctgattcttttctgtcaagctcttga 60 ggtggacatttccctccaagggcctgggattgtaccaggaagaagtgaggtttccctgag 120 tctccaggggcctagaggtggaggctgcttccccattgctacaggggccccttttattgt 180 cctcctgcccctgggtctctacctggtctttcacctctgttgcttctttgggctcttctg 240 cgctcacctccgtcttcgggagcctggctgggatcacctgatcatctaatgaaggaagtt 300 gaaggttaaacttgcctctgagacaagggatccttacggggctgaggtgtccaaacatta 360 tggagttgtgaggagacagcacgggtttcttccttgagggggggctccagaccacaggac 420 gcaggaccctctgtggggtgcccgtgttccgagggataagacacagcctcataggggcgc 480 cgtcccacctgactggaaaagaaggcccaagatgtcgctgacggttgaagaggagtggga 540 aacggcccacaattccccgggcaggcacaggtgcaggagctgcagggtgagcccggccag 600 ctgggaaggcctcacggacaagacgagcaggttgccgatggcatggccaggacctgcggc 660 agaaccaggaacaaaatacgcttagcgagttgcccattttgagtgagttgtgcacagacg 720 aaactaagggtcagaagcggagaggatactcctaagtcacccacttctctgtggctgggt 780 gcacactgggcatctgggagtttatgacatcactatggggctggtgacagagccagtgtg 840 tggaggagtgcttaggagcccagcgagggtgcctacaagaggagtcaaagggcaaagggt 900 gagacccttccaccggtccagctggactctagcctcagggatatcctgctcctgggggca 960 agtgtgtggccctggatgggcccccctgtggggctgttgggggtgcgaggctgatccgcc 1020 agagcccttccacctggcgcctggcccaggtgctggctagcacccagtggccctgttttg 1080 gccggccctgtcccccaggttacagggccagaacctggaagcagagcgcaggaccagcca 1140 gatcgcgccaggcttccccggggcctctccagtgcctctgtgccacctggagccaggccc 1200 gccttctccatggctgccgtggcctcaagggccaccagccttgctccgcaggttccaaag 1260 agaggacgcagtgccctgacctgactggatgcacctcttaccacatgcctccctggcagg 1320 cagggtctccactttttacaaatttgcctgagaccattcctcaggtcattcaggtggtca 1380 tggcccagccaggctttgaacccaggctgtgcgattccacagctggcgctctggcctgtg 1440 tgcctcatgatcatggatacagcatctattcttattttttcatgtagtcctggggtactt 1500 agcaccgtggcatatctgtaataagcacatgcacacctcgaaggaggtcttcacttcaac 1560 atacgagttgaccatggcatgctctgggctccagtcctctacaaagacgtagggcaggaa 1620 ctaccagttgtcaggacaggaccatcccacattgctcttctaatggagcctttcacccca 1680 gatgttctttctcgtctgatgggaaggatccaagtatgtaaagattatgttatagatcag 1740 ctttggtctgtcctaaaagaaatttgccagtggattattccatatggataaaagtcagtt 1800 tctctggtcttcctggaatgtgtctagaaagcaaatagattatttacaagttcatagtag 1860 atcaatgtattggattaaaatatgacaaacataatttggtcattgtgagcatgccagctt 1920 ggtcaactattcaccacacatgatgccctaaatataactctaggttttcttatgcccaag 1980 agagggacatactcttgggtgtctggactagggaaacatgtatgaaaaaccatttggcca 2040 ctctacatcttgttattggagaattgaaaccatctatattcaaagatattattaaaaggc 2100 aagaagttaaaaaataaaaaaataaattgcatgtctgttgttatggaattttataactgt 2160 acataccttttatttaatccattcacaggtcatattataggatatctttcatttaataag 2220 tatactaaatccataccttatactgttgagttttgatagtaccagctcgtgccgaattcg 2280 atatcaagcttatcgataccgtcgac 2306 <210> 59 <211> 391 <212> DNA
<213> Homo sapiens <400>

gactgcatgcacctgggccttgtcagtctgattcttttctgtcaagctcttgaggtggac 60 atttccctccaagggcctgggattgtaccaggaagaagtgaggtttccctgagtctccag 120 gggcctagaggtggaggctgcttccccattgctacaggggccccttttattgtcctcctg 180 cccctgggtctctacctggtctttcacctctgttgcttctttgggctcttctgcgctcam 240 ctccgtcttcgggagcctggctgggatcacctgatcatctaatgaaggaagttgaaggtt 300 aaamttgcctctgagacaagggrtcccttacggggctgaggtgtccaaacattatggagt 360 tgtgaggagacagcacgggtttcttccttga 391 <210> 60 <211> 769 <212> DNA
<213> Homo sapiens <400>

ctcgaggtcgacggtatcgataagcttgatatcgaattcggcacgagctggtactatcaa 60 aactcaacagtataaggtatggatttagtatacttattaaatgaaagatatcctataata 120 tgacctgtgaatggattaaataaaaggtatgtacagttataaaattccataayaacagac 180 atgcaatttatttttttattttttaacttcttgccttttaataatatctttgaatataga 240 tggtttcaattctccaataacaagatgtagagtggccaaatggtttttcatacatgtttc 300 cctagtccagacacccaagagtatgtccctctcttgggcataagaaaacctagagttata 360 tttagggcatcatgtgtggtgaatagttgaccaagctggcatgctcacaatgaccaaatt 420 atgtttgtcatattttaatccaatacattgatctactatgaacttgtaaataatctattt 480 gctttctagacacattccmggaagaccagagaaactgacttttatccatatggaataatc 540 cactggcaaatttcttttaggacagaccaaagctgatctataacataatctttacatact 600 trgatccttcccatcagacgagaaagaacatctggggtgaaaggctccattagaagagca 660 atgtgggatggtgctgtgctgacaactggtagttcctgccctacgtctttgtagaggact 720 ggagcccaga gcatgccatg gtcaactcgt atgttgaagt gaagacctc 769 <210> 61 <211> 2338 <212> DNA
<213> Homo sapiens <220>
<221> SITE
<222> (4) <223> n equals a,t,g, or c <400>

ctcnctatggggcgaatgggkaccggccsccctttttttttttttttttaagattccagc 60 tttttttattttttccccttttacacaaaacaaagtagaagaaataatacaggattaaaa 120 ctgcaaaagtagttaatcagtagaatatgtatgcacacaaaaaatccagataggaagaca 180 ggcttatttacaatgaaagtaaggtaaaattaacgtagttttcttatgaaataataaaaa 240 aaaatcaaacatatgctacaatgggcaccactgttcacagcaatattaaagaggagaaaa 300 caacacttatttaatagggacagatataccacaaggtacaacatcagtgcaataaattca 360 caaaactatattacagctagttaatcagtttaagaattgttcccgtcagtcacatttttt 420 ggccctcagaagttcattcctaaagatttcaactactctaaatttctagctaccaagaag 480 ttaagaatgattataagaagctttccaaggagttatgaaatctttgtagaccagaggcca 540 actatcatcacctcaagtctgctctcaccaacagcccttgtatttttcagggagaaatct 600 ctaggaaaaaagtcagacaccagtgtagtcactatctcccatgtcaaacctaggggacta 660 aaatggtcagtattaccataaaatgataattttgaggtttaccttaaaaggcttattctg 720 gtctcaaaaattagataagattatcttctactgaaatgaatattgactaaacatagaaga 780 ctgctgtccttttgccagtcttagagtgaaagtcatagacatgagtcttaacctactgta 840 tactatagctaatgtcagctgaaaatctgaaattaaaagcatgctagaaatcctaaatgc 900 aatcttttggaagtctgctattaaaaagcctttaaggatttactaacttcgagtctaagt 960 gcaaggggacgaaagcttaagcctgtcagacattccttttcttggacaaaaagatcaaag 1020 tttcctacaaattgctaagctttgcacaagggagaagcctacatgtactagtgcatggaa 1080 tcagtttcatcttatttcatggggactcttctcccactggaaagaaacagaatgaggaat 1140 gaatcttaattggtctcttcatcagaagtggtaaacttggtctctatattcacgaagtca 1200 gacagttttttaagcagactgtggaagcagacagaaccagcttcctgtagccacagacca 1260 ctacatggtatctaagctaaagcaaagatgaacaattatccagattcacttgaactgtac 1320 taaagggcaaggttcaccactacaaaaaggaagttgtctaaaagcaagaattcaattaac 1380 gctgggtaagaaaagtcaaaacactaatgagttgtccatgaagccaactgctaagaacgc 1440 gctcaactatacgcgacatgaagacactatgcacgaagccttacttggcgagtctgaatt 1500 tctattaactaagggcagagtgagggagaacaaagagctacttccgtaacattttagtat 1560 ccagatagtacagcagaaacggttcccggggcaatgggtgctgcattaatcacactgatt 1620 aaagcagatgaatcattcgtttttcttttctttttgtttgagaagtttgtttatctccct 1680 cttggccattccaatgtacttcaaaatgattccatgttggtttagtccaggaagcaatag 1740 taaggaagtcactatcaggtaggtgagaagcaggttgtggacttgttgtcccacccaagc 1800 aaccgcagcaagggaaacgatcatggtcatgaagtacatcttaggtttttcttcctttag 1860 tgtgaagaggcgtttccaccaacccacagctctgcgtcgagtttttactagattgctgca 1920 aatttcatggaatctttgctgttgttcagtggtccatttattggagccaaaaattctagg 1980 cgctagaatgggaacaaggtagtcagccaagcacaaaaacataacaaaacaggaaacgcc 2040 ggacagaacagatggatctagatagtagataatcagaaacaccaaagaaaccacacccat 2100 gatggcaggtggaaaccaggctctttcccatcggaggactttatcagccatcagcatcac 2160 ttctccccatccttgcagctgttcttccagacttgcagtctctgcagccagcaggttggt 2220 gctgcgattatctccctccgccatcgtctcggggatgcagtctctacaagcgcaggccac 2280 ctccccaacgagtcctccaaccgaaacccgcaccggacgcgtgggcggacgcgtgggc 2338 <210> 62 <211> 560 <212> DNA

<213> Homo Sapiens <220>
<221> SITE
<222> (517) <223> n equals a,t,g, or c <400> 62 tcgagtttttttttttttttttcattattgtttttattttgatgatgtgatgattactgc 60 atttcaaggccaaggctattttattgactgcaacctattaaaatgtaagagagtaataaa 120 aaaaagagaataatattaattaattcagactatatctctacatagagctggaaaaatatt 180 ataaaaaatatattttgcagttaaattgagtatctttcatatagataagctagatcacat 240 atatttaatgaagcaatataactagcattattcactagtagttctgttaccaaatacaat 300 cttaagtaatttaatgaattccagtgtcatcagtctggtgacattttaaaatttaaattc 360 caatacactttgattacagagtccaaaattatctttgctactaaaaaataacattttata 420 tcataagagcacgtctgtgataaacattgaaacgtgtattattgaaaaggagttattctt 480 agcattttctcctacgtgatctaccctttcttattcntctcgttctcttaacatattcct 540 tktaaawtaacgtagactat 560 <210> 63 <211> 685 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (303) <223> n equals a,t,g, or c <400> 63 cggcacgagcggcacgagcccacagcctgcaggaccgaggcccctgtgtgagggactggg 60 ggtgtgaggagttgccaggagaagcagaccctgccaggtgcccatctcctctcagcaccc 120 ctccccccaacacacttcctgtaatggcgtctccaatggggagcccattagtcctgctta 180 gggcagcacctaagtactgtggctgataaatataacatgagcccatcgattagaatcctc 240 ctagtcctacagcagctcggctccctaatggctcccctcccctcagcacactgaytaccc 300 acntctcaccccccamcccaggctgattttctggtttacttgaggctgtggggcaaacat 360 gagaggagcagaagtgaggtgatgggtgcaacacccagagcccagtcaaaggggccactg 420 cttggagacaggctggactggtytgggccctgaggcagcctggaagcccctcggtcagtt 480 aggggcagctaggggaggaccgcctgtgagttcccggggcccggcctgcggtargacatg 540 garcaaggaargcagarggccctactttytgtgrrgcscaagacakcactgcccttgggg 600 gccttgggggtctcctggtaatctcaaaaaaactgcaatgatccactctggcctgttacc 660 acgcctcttctcctgggattccttg 685 <210> 64 <211> 604 <212> DNA
<213> Homo Sapiens <400> 64 ccgttagacg ccagttcgaa attaaccctc actaaaggga acaaaagctg gagctccacc 60 gcggtggcgg ccgctctaga actagtggat cccccgggct gcaggaattc ggcacgagca 120 gcgcgcactt accaccatgc agatccaggt cgcggggctg ctgcagtttg ccgtgcccct 180 cttctccact gcagaggaag acctgcttgc aatccagctc ctgctgaact cctcagagtc 240 cagccttcac cagctgaccg ccatggtgga ctgccgaggg ctgcacaagg attatctgga 300 cgctcttgct ggcatctgct acgacggcct ccagggcttg ctgtaccttg gcctcttctc 360 cttcctggcc gccctcgcct tctccaccat gatctgtgca gggccaaggg cctggaagca 420 cttcaccacc agaaacagag actacgatga cattgatgat gatgacccct ttaaccccca 480 agcctggcgc atggcggctc acagtccccc gaggggacag cttcacagct tctgcagcta 540 cagcagtggc ctgggaagtc agaccagcct gcagcccccg gcccagacca tctccaacgc 600 ccct 604 <210> 65 <211> 1520 <212> DNA

<213> Homoapiens s <220>

<221> SITE

<222> (716) <223> n ls a,t,g,or c equa <220>

<221> SITE

<222> (748) <223> n or c equals a,t,g, <220>

<221> SITE

<222> (1398) <223> n or c equals a,t,g, <220>

<221> SITE

<222> (1410) <223> n or c equals a,t,g, <220>

<221> SITE

<222> (1424) <223> n or c equals a,t,g, <220>

<221> SITE

<222> (1432) <223> n or c equals a,t,g, <400> 65 tcgacccacgcgtccgctcagagaatgtccagagctaagtctgagcaaagggccagcttt 60 tgcttttggggctggggtacaggcagattcctgttggagagcatttattttacttttcaa 120 gtgtgggctttctggaaggtaatgggaaactgcaaggtgtcttgcaagcaggtgtttggc 180 ttatggtgatgtggttcccagggtggcttttgggcaggggcagggcggggtcctccagga 240 gctgtctgtcggtgctgagctcagtctgcttccctaccactcagcccactgcttggatgg 300 ttccctctgtcactctcatcttacactgtccaggttttagcactgaaagtcatatgtgtg 360 ggaagcctctcagtcccaggccaaccagaacagttggtcgccccgtgtctaacatacctg 420 tctaactgtctccagggcatggctcatcagcgtggctgtggcaccctctccctgccttct 480 cttgtctccttcactgtcaagtctgggaaagggctcctggtgggtgtccgatgaggcaca 540 gctgggtgacaaagatggctccagctctggttggggcgtgcacctcatggtgacacctcc 600 ctctgtgggacctccgagggtcaaggaaggaggcaccttccatcctaggtttaggattca 660 ttgcacttacacaggattccccgtggcagatcacctgtgttccagargctgtgtanttca 720 aggtgtgtgtgaggcagttgcatggtangggggcacctataactgggggtaagtctgtaa 780 tggtgcaatagatgatgttagaacagtgggggtccccctccctctcaggcctcaaaagca 840 caaccaccagarctggtgggtggcactgggccatggggagacctgaccagcaaggctccc 900 tctttcctctgtccaggccacccagcctgaratacagtgctggtcactctaaggaatgct 960 gtggcccccaagggacaccccttgggtggctcctcccwggcttctcactgckggctggca 1020 rgcargctggtgggcgggcaarcgtaatattacccctaagataaatgaagaagagtttca 1080 gcccttattggattatagtttttattgaactttataaattccaattagcagacccctggg 1140 caataagcgacacacagcccgtggctcgctttatagcgtgattatgaccctgtttatcct 1200 cccctgaacgagaagtattaatggaggaaacaaaggaaagggactcaaaaatacataaaa 1260 acagatattggtgacacagagataaacagacaggagtcagggagagaaaacaatctccgg 1320 gcaggcaagggggaacagagaatcagagattctaagaacccatgtgctcagaccaccgtg 1380 agcttggatgtccttctnggctcctcccanggcacaaacccctnagctcctnaagctgct 1440 ggggctgaagagacctccctcccaagcctactcaggatcttgctggggtggatgtgatta 1500 taatgaaatcaggtgccacc 1520 <210> 66 <211> 596 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (595) <223> n equals a,t,g, or c <400> 66 ggccgcccttttttttttttttttaatcttccactgaagcctccaagaaagatgacctgc 60 agaagggggaggaaggcgcaaacaaagccagacaaagtgaggagggcacgccatccttga 120 gcacacttctttctcctgggagatgcagcaagtcgggcaagggtttcagtccagaagagc 180 agtaccaccacccacctggctggctcaggaccccccagctcacccccacacctctctagg 240 gacagcttcttcccagctgctgttctgggtcatggctgttagaaatttaaagctgatgaa 300 cgcaaaccagagtatccaggaagaaatgaaaagcaggtggggttggagggcattcccagc 360 tgtggccaggcccttggaggctgcccatctgtctgtgggctctcaggwyctccaggtgga 420 agctcctgggagacagatcttacttggttcaattcaaactcaaggattcttaatcttatg 480 ctttcagtcccctggaggarggggcagaatgcatgatcagttttcagagcttggacaacc 540 ccctgtacctataggcaaaattacttgtgcgtgcatgtgtacataatgcatgtgna 596 <210> 67 <211> 2093 <212> DNA
<213> Homo Sapiens <220>
<221> SITE
<222> (2080) <223> n equals a,t,g, or c <400> 67 ggctaggtggcgcgcgggtccggcgggcggttggcttgagcgggaccggagctgaggcag 60 gaagagccggcgccatggtggagaaggaggaggctggcggcggcattagcgaggaggagg 120 cggcacagtatgaccggcagatccgcctgtggggactggaggcccagaaacggctgcggg 180 cctctcgggtgcttcttgtcggcttgaaaggacttggggctgaaattgccaagaatctca 240 tcttggcaggagtgaaaggactgaccatgctggatcacgaacaggtaactccagaagatc 300 ccggagctcagttcttgattcgtactgggtctgttggccgaaatagggctgaagcctctt 360 tggagcgagctcagaatctcaaccccatggtggatgtgaaggtggacactgaggatatag 420 agaagaaaccagagtcatttttcactcaattcgatgctgtgtgtctgacttgctgctcca 480 gggatgtcatagttaaagttgaccagatctgtcacaaaaatagcatcaagttctttacag 540 gagatgtttttggctaccatggatacacatttgccaatctaggagagcatgagtttgtag 600 aggagaaaactaaagttgccaaagttagccaaggagtagaagatgggcccgacaccaaga 660 gagcaaaacttgattcttctgagacaacgatggtcaaaaagaaggtggtcttctgccctg 720 ttaaagaagccctggaggtggactggagcagtgagaaagcaaaggctgctctgaagcgca 780 cgacctccgactactttctccttcaagtgctcttaaagttccgtacagataaaggaagag 840 atcccagttctgatacatatgaggaagattctgagttgttgctccagatacgaaatgatg 900 tgcttgactcactgggtattagtcctgacctgcttcctgaggactttgtcaggtactgct 960 tctccgagatggccccagtgtgtgcggtggttggagggattttggcacaggaaattgtga 1020 aggccctgtctcagcgggaccctcctcacaacaacttcttcttcttcgatggcatgaagg 1080 ggaatgggattgtggagtgccttggccccaagtgaactcaagatttggcagccccagaga 1140 tgccaactgcagcatgcccacctgtattccctgtccccttccttcatgaaggcatctcca 1200 ggcaaggaaaactgaagtcattggcccgatacaaaacatttcctgcaacgaaggaggtgg 1260 tgccgacgtgctgcttcccatcaccagcagctgctcgacaaggggcgcagggtggctgtc 1320 tttgttccagcactgttcaggctgcctgtcatcccgggcctgccagctcccctgagtgat 1380 gagcacttccaagcacccctctgccctttctctgtccttatgctgtcccggcctcgccag 1440 ccctctggggcattgtgggagatgcctgccaggaatgagcaagctctgttgctcgggagc 1500 ctcttgtcaccttcttggacttattccccacctgataccttatagagaaaagtgtgaatt 1560 caggtggagagtaggcccaggccccatgaggcaccagtggaagcacagctccaagttcag 1620 acaggtgcccttagagaggaaaaccatgacaggcaaatgcatttcctctggagtttgaga 1680 ccctgacaaacaacaggtggcatctggtgtgctgttcttgagttttcgtttaggattagt 1740 tgagttccagctgggttttgggagaaaggagatgctaccaagtcttggatgttagggcga 1800 gaccctgcaagttgagtattagagagcttgtctttcaaggcaggttcctggggcttcagg 1860 gctaggagggaggagcctgcccttttaacagaaccccagtcacatgcggctcaagtcact 1920 cagaggctgttgcatttcagggctatgttggtcctttgtttacctcctaaaccacagctg 1980 tttgtgtttcacatatgttgtgaattttccttggttctttttaaaggaatgataataaag 2040 ttacttgctttaggaaaaaaaaaaaaaaaaatcgccgganggggggggccccg 2093 <210> 68 <211> 261 <212> PRT
<213> Homo Sapiens <400> 68 Met Ala Val Thr Ala Cys Gln Gly Leu Gly Phe Val Val Ser Leu Ile Gly Ile Ala Gly Ile Ile Ala Ala Thr Cys Met Asp Gln Trp Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala Val Phe Asn Tyr Gln Gly Leu Trp Arg Ser Cys Val Arg Glu Ser Ser Gly Phe Thr Glu Cys Arg Gly Tyr Phe Thr~Leu Leu Gly Leu Pro Ala Met Leu Gln Ala Val Arg Ala Leu Met Ile Val Gly Ile Val Leu Gly Ala Ile Gly Leu Leu Val Ser Ile Phe Ala Leu Lys Cys Ile Arg Ile Gly Ser Met Glu Asp Ser Ala Lys Ala Asn Met Thr Leu Thr Ser Gly Ile Met Phe Ile Val Ser Gly Leu Cys Ala Ile Ala Gly Val Ser Val Phe Ala Asn Met Leu Val Thr Asn Phe Trp Met Ser Thr Ala Asn Met Tyr Thr Gly Met Gly Gly Met Val Gln Thr Val Gln Thr Arg Tyr Thr Phe Gly Ala Ala Leu Phe Val Gly Trp Val Ala Gly Gly Leu Thr Leu Ile Gly Gly Val Met Met Cys Ile Ala Cys Arg Gly Leu Ala Pro Glu Glu Thr Asn Tyr Lys Ala Val Ser Tyr His.Ala Ser Gly His Ser Val Ala Tyr Lys Pro Gly Gly Phe Lys Ala Ser Thr Gly Phe Gly Ser Asn Thr Lys Asn Lys Lys Arg Tyr Asp Gly Gly Ala Arg Thr Glu Asp Glu Val Gln Ser Tyr Pro Ser Lys His Asp Tyr Val <210> 69 <211> 27 <212> PRT
<213> Homo sapiens <400> 69 Trp Ser Pro Leu Val Trp Leu Trp Pro Met Val Trp Thr Phe Thr Lys Leu Glu Ser Thr His Pro Ser Ser Leu Thr Phe <210> 70 <211> 149 <212> PRT
<213> Homo Sapiens <400> 70 Met Val Leu Leu Trp Ala Ser Val Leu Phe Pro Ala Pro Glu Asp Trp Ala Glu Leu Gln Gly Ala Val Tyr Arg Leu Leu Val Val Leu Leu Cys Cys Leu Ala Thr Arg Lys Leu Pro His Phe Leu His Pro Gln Arg Asn Leu Leu Gln Gly Ser Gly Leu Asp Leu Gly Ala Ile Tyr Gln Arg Val Glu Gly Phe Ala Ser Gln Pro Glu Ala Ala Leu Arg Ile His Ala Thr His Leu Gly Arg Ser Pro Pro Pro Arg Ile Gly Ser Gly Leu Lys Ala Leu Leu Gln Leu Pro Ala Ser Asp Pro Thr Tyr Trp Ala Thr Ala Tyr Phe Asp Val Leu Leu Asp Lys Phe Gln Val Phe Asn Ile Gln Asp Lys Asp Arg Ile Ser Ala Met Gln Ser Ile Phe Gln Lys Thr Arg Thr Leu Gly Gly Glu Glu Ser <210> 71 <211> 547 <212> PRT
<213> Homo sapiens <400> 71 Met Ala Met Gly Leu Phe Arg Val Cys Leu Val Val Val Thr Ala Ile Ile Asn His Pro Leu Leu Phe Pro Arg Glu Asn Ala Thr Val Pro Glu Asn Glu Glu Glu Ile Ile Arg Lys Met Gln Ala His Gln Glu Lys Leu Gln Leu Glu Gln Leu Arg Leu Glu Glu Glu Val Ala Arg Leu Ala Ala Glu Lys Glu Ala Leu Glu Gln Val Ala Glu Glu Gly Arg Gln Gln Asn Glu Thr Arg Val Ala Trp Asp Leu Trp Ser Thr Leu Cys Met Ile Leu Phe Leu Met Ile Glu Val Trp Arg Gln Asp His Gln Glu Gly Pro Ser Pro Glu Cys Leu Gly Gly Glu Glu Asp Glu Leu Pro Gly Leu Gly Gly Ala Pro Leu Gln Gly Leu Thr Leu Pro Asn Lys Ala Thr Leu Gly His Phe Tyr Glu Arg Cys Ile Arg Gly Ala Thr Ala Asp Ala Ala Arg Thr Arg Glu Phe Leu Glu Gly Phe Val Asp Asp Leu Leu Glu Ala Leu Arg Ser Leu Cys Asn Arg Asp Thr Asp Met Glu Val Glu Asp Phe Ile Gly Val Asp Ser Met Tyr Glu Asn Trp Gln Val Asp Arg Pro Leu Leu Cys His Leu Phe Val Pro Phe Thr Pro Pro Glu Pro Tyr Arg Phe His Pro Glu Leu Trp Cys Ser Gly Arg Ser Val Pro Leu Asp Arg Gln Gly Tyr Gly Gln Ile Lys Val Val Arg Ala Asp Gly Asp Thr Leu Ser Cys Ile Cys Gly Lys Thr Lys Leu Gly Glu Asp Met Leu Cys Leu Leu His Gly Arg Asn Ser Met Ala Pro Pro Cys Gly Asp Met Glu Asn Leu Leu Cys Ala Thr Asp Ser Leu Tyr Leu Asp Thr Met Gln Val Met Lys Trp Phe Gln Thr Ala Leu Thr Arg Ala Trp Lys Gly Ile Ala His Lys Tyr Glu Phe Asp Leu Ala Phe Gly Gln Leu Asp Ser Pro Gly Ser Leu Lys Ile Lys Phe Arg Ser Gly Lys Phe Met Pro Phe Asn Leu Ile Pro Val Ile Gln Cys Asp Asp Ser Asp Leu Tyr Phe Val Ser His Leu Pro Arg Glu Pro Ser Glu Gly Thr Pro Ala Ser Ser Thr Asp Trp Leu Leu Ser Phe Ala Val Tyr Glu Arg His Phe Leu Arg Thr Thr Leu Lys Ala Leu Pro Glu Gly Ala Cys His Leu Ser Cys Leu Gln Ile Ala Ser Phe Leu Leu Ser Lys Gln Ser Arg Leu Thr Gly Pro Ser Gly Leu Ser Ser Tyr His Leu Lys Thr Ala Leu Leu His Leu Leu Leu Leu Arg Gln Ala Ala Asp Trp Lys Ala Gly Gln Leu Asp Ala Arg Leu His Glu Leu Leu Cys Phe Leu Glu Lys Ser Leu Leu Gln Lys Lys Leu His His Phe Phe Ile Gly Asn Arg Lys Val Pro Glu Ala Met Gly Leu Pro Glu Ala Val Leu Arg Ala Glu Pro Leu Asn Leu Phe Arg Pro Phe Val Leu Gln Arg Ser Leu Tyr Arg Lys Thr Leu Asp Ser Phe Tyr Glu Met Leu Lys Asn Ala Pro Ala Leu Ile Ser Glu Tyr Ser Leu His Val Pro Ser Asp Gln Pro Thr Pro Lys Ser <210> 72 <211> 245 <212> PRT
<213> Homo sapiens <400> 72 Met Leu Gln Ala Arg Asn Gln Ser Pro Ser Ser Gln Arg Pro Leu Asp Val Leu Arg Arg Asn Gln Asp Pro Gln Ser Pro Ala Ser Ile Ser Val Ile Ile Phe Ile Thr Pro Lys Glu Glu Pro Ala Leu Gln Glu Gly Leu His Leu Gln Glu Asp Gly Leu Pro Ala Thr Ala Glu Asp Ala Ala Thr Cys Leu Thr Val Leu Ser Ser Gln Pro Ala Ser Cys Arg Ala Ser Cys Cys Leu Arg Ala Asp Gly Pro Gly Met Leu Ala His Thr Cys Glu His Ser Thr Gly Lys Trp Glu His Ser Thr Arg Lys Trp Glu His Ser Thr Gly Lys Trp Glu His Ser Thr Gly Lys Trp Gly Leu Thr Ala Leu Gln Asn Gly Ser Thr Val Leu Gly Asn Gly Ser Thr Val Leu Gly Ser Gly Ser Thr Val Leu Arg Ser Gly Ser Thr Val Leu Arg Asn Gly Ser Thr Leu Leu Arg Asn Gly Ser Thr Val Leu Gly Asn Gly His Thr Val Leu Gly Asn Gly His Thr Val Leu Arg Asn Gly Ser Thr Val Leu Gly Asn Gly Ser Thr Val Leu Gly Asn Gly Ser Pro Gln Tyr Trp Glu Arg Gly Val His Ser Thr Arg Lys Trp Glu His Ser Thr Gly Lys Trp Glu His Ser Thr Gly Lys Trp Glu His Ser Thr Gly Lys Pro Gln Thr Trp Ile Leu Ser Phe Ser Ala <210> 73 <211> 434 <212> PRT
<213> Homo sapiens <400> 73 Met Gly Met Gly Arg Gly Ala Gly Arg Ser Ala Leu Gly Phe Trp Pro Thr Leu Ala Phe Leu Leu Cys Ser Phe Pro Ala Ala Thr Ser Pro Cys Lys Ile Leu Lys Cys Asn Ser Glu Phe Trp Ser Ala Thr Ser Gly Ser His Ala Pro Ala Ser Asp Asp Thr Pro Glu Phe Cys Ala Ala Leu Arg Ser Tyr Ala Leu Cys Thr Arg Arg Thr Ala Arg Thr Cys Arg Gly Asp Leu Ala Tyr His Ser Ala Val His Gly Ile Glu Asp Leu Met Ser Gln His Asn Cys Ser Lys Asp Gly Pro Thr Ser Gln Pro Arg Leu Arg Thr Leu Pro Pro Ala Gly Asp Ser Gln Glu Arg Ser Asp Ser Pro Glu Ile Cys His Tyr Glu Lys Ser Phe His Lys His Ser Ala Thr Pro Asn Tyr Thr His Cys Gly Leu Phe Gly Asp Pro His Leu Arg Thr Phe Thr Asp Arg Phe Gln Thr Cys Lys Val Gln Gly Ala Trp Pro Leu Ile Asp Asn Asn Tyr Leu Asn Val Gln Val Thr Asn Thr Pro Val Leu Pro Gly Ser Ala Ala Thr Ala Thr Ser Lys Leu Thr Ile Ile Phe Lys Asn Phe Gln Glu Cys Val Asp Gln Lys Val Tyr Gln Ala Glu Met Asp Glu Leu Pro Ala Ala Phe Val Asp Gly Ser Lys Asn Gly Gly Asp Lys His Gly Ala Asn Ser Leu Lys Ile Thr Glu Lys Val Ser Gly Gln His Val Glu Ile Gln Ala Lys Tyr Ile Gly Thr Thr Ile Val Val Arg Gln Val Gly Arg Tyr Leu Thr Phe Ala Val Arg Met Pro Glu Glu Val Val Asn Ala Val 275 ' 280 285 Glu Asp Trp Asp Ser Gln Gly Leu Tyr Leu Cys Leu Arg Gly Cys Pro Leu Asn Gln Gln Ile Asp Phe Gln Ala Phe His Thr Asn Ala Glu Gly Thr Gly Ala Arg Arg Leu Ala Ala Ala Ser Pro Ala Pro Thr Ala Pro Glu Thr Phe Pro Tyr Glu Thr Ala Val Ala Lys Cys Lys Glu Lys Leu Pro Val Glu Asp Leu Tyr Tyr Gln Ala Cys Val Phe Asp Leu Leu Thr Thr Gly Asp Val Asn Phe Thr Leu Ala Ala Tyr Tyr Ala Leu Glu Asp Val Lys Met Leu His Ser Asn Lys Asp Lys Leu His Leu Tyr Glu Arg Thr Arg Asp Leu Pro Gly Arg Ala Ala Ala Gly Leu Pro Leu Ala Pro Arg Pro Leu Leu Gly Ala Leu Val Pro Leu Leu Ala Leu Leu Pro Val Phe Cys <210> 74 <211> 94 <212> PRT
<213> Homo Sapiens <400> 74 Met Leu Leu Gly Phe Leu Val Leu Ile Pro Trp Gly Ser Leu Ile Leu Gly Ser Ser Asp Leu Asp Pro Ser Ser Leu Pro Leu Gly Thr Arg Gly His Gly Trp Arg Trp Pro Pro Leu Ser Pro Val Gln Ile Leu Tyr Pro Leu Ala Gly Asp Pro His Ala Ala Val Ser Cys Ser Cys Cys Gly Glu Thr Glu Leu Arg Ala Leu Leu Thr Gly Ser Leu Pro Met Glu Ala Phe Ser Gly Leu His Ser Ile Glu Tyr Ser Ser Arg Thr Ala Cys <210> 75 <211> 135 <212> PRT
<213> Homo Sapiens <400> 75 Met Leu His Leu Lys Ala Leu Gly Arg Gly Leu Leu Gln Ala Cys Leu Leu Ala Thr Gly Gly Ser Met Ala Cys Asp Ser Ile Thr Pro Ile Phe Thr Gln His Ser Pro Arg Val Pro Leu Ser Pro Ser Phe Pro Leu Leu Gln Glu Thr Gly Ala Phe Ala Pro Ser Pro Arg Leu Glu Gly Ser Gly Met Ile Leu Ala His Cys Cys Leu Glu Leu Leu Cys Ser Ser Asp Pro Pro Ala Ser Ala Ile Gln Asn Cys Trp Asp Tyr Arg Cys Glu Pro Arg His Leu Ala Ser Ile Ser Ser Phe Tyr Lys Asp Ile Gly Arg Ile Gly Leu Gly Pro Val Leu Leu Gln Tyr Asn Leu Ile Leu Val Asn Tyr Ile Cys His Asn Ser Val Ser Lys <210> 76 <211> 280 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (94) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (95) <223> Xaa equals any of the naturally occurring L-amino acids <400> 76 Met His Ser Gln Cys Gln Gly Phe Phe Ser Ser Leu Thr Met Leu Asn Pro Leu Arg Gln Leu Phe Lys Leu Met Ala Ser Leu Phe Leu Ser Val Phe Thr Leu Gly Leu Pro Phe Ala Leu Phe Gln Tyr Tyr Ala Tyr Thr Gln Phe Cys Leu Pro Gly Ser Ala Arg Pro Ile Pro Glu Pro Leu Val Gln Leu Ala Val Asp Lys Gly Tyr Arg Ile Ala Glu Gly Asn Glu Pro Pro Trp Cys Phe Trp Asp Val Pro Leu Ile Tyr Ser Tyr Xaa Xaa Asp Val Tyr Trp Asn Val Gly Phe Leu Lys Tyr Tyr Glu Leu Lys Gln Val Pro Asn Phe Leu Leu Ala Ala Pro Val Ala Ile Leu Val Ala Trp Ala Thr Trp Thr Tyr Val Thr Thr His Pro Trp Leu Cys Leu Thr Leu Gly Leu Gln Arg Ser Lys Asn Asn Lys Thr Leu Glu Lys Pro Asp Leu Gly Phe Leu Ser Pro Gln Val Phe Val Tyr Val Val His Ala Ala Val Leu Leu Leu Phe Gly Gly Leu Cys Met His Val Gln Val Leu Thr Arg Phe Leu Gly Ser Ser Thr Pro Ile Met Tyr Trp Phe Pro Ala His Leu Leu Gln Asp Gln Glu Pro Leu Leu Arg Ser Leu Lys Thr Val Pro Trp Lys Pro Leu Ala Glu Asp Ser Pro Pro Gly Gln Lys Val Pro Arg Asn Pro Ile Met Gly Leu Leu Tyr His Trp Lys Thr Cys Ser Pro Val Thr Arg Tyr Ile Leu Gly Tyr Phe Leu Thr Tyr Trp Leu Leu Gly Leu Leu Leu His Cys Asn Phe Leu Pro Trp Thr <210> 77 <211> 105 <212> PRT
<213> Homo Sapiens <400> 77 Met Phe Ala Leu Ala Trp Lys Val Ile Phe Ser Val Met Leu Gln Asn Pro Ile Arg Tyr Pro Ser Val Leu Gly Ile Lys Ser Ser Leu Leu Ser Ser Leu Val Leu Val Met Val Trp Gly Asn Glu Lys Ser Gly Pro Cys Pro Thr Pro Lys Ser Arg Lys Gly Arg Arg Ser Cys Pro Ala Gln Val Gly Arg Gly Glu Glu Gly Ser Tyr Trp Asp Pro Glu Phe Arg Leu Ser Arg Lys Ser Asn Gln Gly Leu Arg Arg Asp Tyr Leu Ser Leu Tyr His Phe Asn Leu His Phe Arg Asp Thr Phe <210> 78 <211> 92 <212> PRT
<213> Homo Sapiens <400> 78 Met Thr Ser Leu Leu Glu Gly Arg Met Val Leu Cys Val Ser Cys Leu Leu Leu Pro Leu Leu Leu Leu Leu Lys His Phe Asn Gly Leu Met Thr Pro Tyr Leu Ala His Asn Val Tyr Cys Pro Ile Glu Tyr Ile Ser Phe Phe Pro Phe His Glu Lys Asn Ile Glu Tyr Ile Ser Ile Trp Phe Ile Phe Asp Ser Phe Lys Phe Ile Tyr Ser Arg Leu Leu Cys Ile Ser Gln Ile Tyr Val Leu Tyr Arg Ala Tyr Thr Leu Pro His <210> 79 <211> 445 <212> PRT
<213> Homo Sapiens <220>

<221> SITE
<222> (147) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (288) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (293) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (332) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (443) <223> Xaa equals any of the naturally occurring L-amino acids <400> 79 Met Leu Trp Phe Ser Gly Val Gly Ala Leu Ala Glu Arg Tyr Cys Arg Arg Ser Pro Gly Ile Thr Cys Cys Val Leu Leu Leu Leu Asn Cys Ser Gly Val Pro Met Ser Leu Ala Ser Ser Phe Leu Thr Gly Ser Val Ala Lys Cys Glu Asn Glu Gly Glu Val Leu Gln Ile Pro Phe Ile Thr Asp Asn Pro Cys Ile Met Cys Val Cys Leu Asn Lys Glu Val Thr Cys Lys Arg Glu Lys Cys Pro Val Leu Ser Arg Asp Cys Ala Leu Ala Ile Lys Gln Arg Gly Ala Cys Cys Glu Gln Cys Lys Gly Cys Thr Tyr Glu Gly Asn Thr Tyr Asn Ser Ser Phe Lys Trp Gln Ser Pro Ala Glu Pro Cys Val Leu Arg Gln Cys Gln Glu Gly Val Val Thr Glu Ser Gly Val Arg Cys Val Xaa His Cys Lys Asn Pro Leu Glu His Leu Gly Met Cys Cys Pro Thr Cys Pro Gly Cys Val Phe Glu Gly Val Gln Tyr Gln Glu Gly Glu Glu Phe Gln Pro Glu Gly Ser Lys Cys Thr Lys Cys Ser Cys Thr Gly Gly Arg Thr Gln Cys Val Arg Glu Val Cys Pro Ile Leu Ser Cys Pro Gln His Leu Ser His Ile Pro Pro Gly Gln Cys Cys Pro Lys Cys Leu Gly Gln Arg Lys Val Phe Asp Leu Pro Phe Gly Ser Cys Leu Phe Arg Ser Asp Val Tyr Asp Asn Gly Ser Ser Phe Leu Tyr Asp Asn Cys Thr Ala Cys Thr Cys Arg Asp Ser Thr Val Val Cys Lys Arg Lys Cys Ser His Pro Gly Gly Cys Asp Gln Gly Gln Glu Gly Cys Cys Glu Xaa Cys Leu Leu Arg Xaa Pro Pro Glu Asp Ile Lys Val Cys Lys Phe Gly Asn Lys Ile Phe Gln Asp Gly Glu Met Trp Ser Ser Ile Asn Cys Thr Ile Cys Ala Cys Val Lys Gly Arg Thr Glu Cys Xaa Asn Lys Gln Cys Ile Pro Ile Ser Ser Cys Pro Gln Gly Lys Ile Leu Asn Arg Lys Gly Cys Cys Pro Ile Cys Thr Glu Lys Pro Gly Val Cys Thr Val Phe Gly Asp Pro His Tyr Asn Thr Phe Asp Gly Arg Thr Phe Asn Phe Gln Gly Thr Cys Gln Tyr Val Leu Thr Lys Asp Cys Ser Ser Pro Ala Ser Pro Phe Gln Val Leu Val Lys Asn Asp Ala Arg Arg Thr Arg Ser Phe Ser Trp Thr Lys Ser Val Glu Leu Val Leu Gly Glu Thr Gly Ser Ala Cys 420 425 . 430 Ser Ser Thr Ser Pro Cys Ala Gly Thr Ala Xaa Ala Ser <210> 80 <211> 273 <212> PRT
<213> Homo sapiens <400> 80 Met Leu Phe Phe Cys Gly Asp Leu Leu Ser Arg Ser Gln Ile Phe Tyr Tyr Ser Thr Gly Met Thr Val Gly Ile Val Ala Ser Leu Leu Ile Ile Ile Phe Ile Leu Ser Lys Phe Met Pro Lys Lys Ser Pro Ile Tyr Val Ile Leu Val Gly Gly Trp Ser Phe Ser Leu Tyr Leu Ile Gln Leu Val Phe Lys Asn Leu Gln Glu Ile Trp Arg Cys Tyr Trp Gln Tyr Leu Leu Ser Tyr Val Leu Thr Val Gly Phe Met Ser Phe Ala Val Cys Tyr Lys Tyr Gly Pro Leu Glu Asn Glu Arg Ser Ile Asn Leu Leu Thr Trp Thr Leu Gln Leu Met Gly Leu Cys Phe Met Tyr Ser Gly Ile Gln Ile Pro His Ile Ala Leu Ala Ile Ile Ile Ile Ala Leu Cys Thr Lys Asn Leu Glu His Pro Ile Gln Trp Leu Tyr Ile Thr Cys Arg Lys Val Cys Lys Gly Ala Glu Lys Pro Val Pro Pro Arg Leu Leu Thr Glu Glu Glu Tyr Arg Ile Gln Gly Glu Val Glu Thr Arg Lys Ala Leu Glu Glu Leu Arg Glu Phe Cys Asn Ser Pro Asp Cys Ser Ala Trp Lys Thr Val Ser Arg Ile Gln Ser Pro Lys Arg Phe Ala Asp Phe Val Glu Gly Ser Ser His Leu Thr Pro Asn Glu Val Ser Val His Glu Gln Glu Tyr Gly Leu Gly Ser Ile Ile Ala Gln Asp Glu Ile Tyr Glu Glu Ala Ser Ser Glu Glu Glu Asp Ser Tyr Ser Arg Cys Pro Ala Ile Thr Gln Asn Asn Phe Leu Thr <210> 81 <211> 503 <212> PRT

<213> Homo sapiens <400> 81 Met Glu Gln Arg His Val Leu Leu Lys Gln Lys Glu Leu Gly Gly Glu Glu Pro Glu Pro Ser Leu Arg Glu Gly Pro Gly Gly Leu Val Met Glu Gly His Leu Phe Lys Arg Ala Ser Asn Ala Phe Lys Thr Trp Ser Arg Arg Trp Phe Thr Ile Gln Ser Asn Gln Leu Val Tyr Gln Lys Lys Tyr Lys Asp Pro Val Thr Val Val Val Asp Asp Leu Arg Leu Cys Thr Val Lys Leu Cys Pro Asp Ser Glu Arg Arg Phe Cys Phe Glu Val Val Ser Thr Ser Lys Ser Cys Leu Leu Gln Ala Asp Ser Glu Arg Leu Leu Gln Leu Trp Val Ser Ala Val Gln Ser Ser Ile Ala Ser Ala Phe Ser Gln Ala Arg Leu Asp Asp Ser Pro Arg Gly Pro Gly Gln Gly Ser Gly His Leu Ala Ile Gly Ser Ala Ala Thr Leu Gly Ser Gly Gly Met Ala Arg Gly Arg Glu Pro Gly Gly Val Gly His Val Val Ala Gln Val Gln Ser Val Asp Gly Asn Ala Gln Cys Cys Asp Cys Arg Glu Pro Ala Pro Glu Trp Ala Ser Ile Asn Leu Gly Val Thr Leu Cys Ile Gln Cys Ser Gly Ile His Arg Ser Leu Gly Val His Phe Ser Lys Val Arg Ser Leu Thr Leu Asp Ser Trp Glu Pro Glu Leu Val Lys Leu Met Cys Glu Leu Gly Asn Val Ile Ile Asn Gln Ile Tyr Glu Ala Arg Val Glu Ala Met Ala Val Lys Lys Pro Gly Pro Ser Cys Ser Arg Gln Glu Lys Glu Ala Trp Ile His Ala Lys Tyr Val Glu Lys Lys Phe Leu Thr Lys Leu Pro Glu Ile Arg Gly Arg Arg Gly Gly Arg Gly Arg Pro Arg Gly Gln Pro Pro Val Pro Pro Lys Pro Ser Ile Arg Pro Arg Pro Gly Ser Leu Arg Ser Lys Pro Glu Pro Pro Ser Glu Asp Leu Gly Ser Leu His Pro Gly Ala Leu Leu Phe Arg Ala Ser Gly His Pro Pro Ser Leu Pro Thr Met Ala Asp Ala Leu Ala His Gly Ala Asp Val Asn Trp Val Asn Gly Gly Gln Asp Asn Ala Thr Pro Leu Ile Gln Ala Thr Ala Ala Asn Ser Leu Leu Ala Cys Glu Phe Leu Leu Gln Asn Gly Ala Asn Val Asn Gln Ala Asp Ser Ala Gly Arg Gly Pro Leu His His Ala Thr Ile Leu Gly His Thr Gly Leu Ala Cys Leu Phe Leu Lys Arg Gly Ala Asp Leu Gly Ala Arg Asp Ser Glu Gly Arg Asp Pro Leu Thr Ile Ala Met Glu Thr Ala Asn Ala Asp Ile Val Thr Leu Leu Arg Leu Ala Lys Met Arg Glu Ala Glu Ala Ala Gln Gly Gln Ala Gly Asp Glu Thr Tyr Leu Asp Ile Phe Arg Asp Phe Ser Leu Met Ala Ser Asp Asp Pro Glu Lys Leu Ser Arg Arg Ser His Asp Leu His Thr Leu <210> 82 <211> 230 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (63) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (66) <223> Xaa equals any of the naturally occurring L-amino acids <400> 82 Met Gly Asn Ser Leu Ser Val Phe Cys Ser Trp Phe Cys Arg Arg Ser Trp Pro Cys His Arg Gln Pro Ala Arg Leu Val Arg Glu Ala Phe Pro Ala Gly Arg Ala His Pro Ala Ala Pro Ala Pro Val Pro Ala Arg Gly Ile Val Gly Arg Phe Pro Leu Leu Phe Asn Arg Gln Arg His Xaa Gly Pro Xaa Phe Pro Val Arg Trp Asp Gly Ala Pro Met Arg Leu Cys Leu Ile Pro Arg Asn Thr Gly Thr Pro Gln Arg Val Leu Arg Pro Val Val Trp Ser Pro Pro Ser Arg Lys Lys Pro Val Leu Ser Pro His Asn Ser Ile Met Phe Gly His Leu Ser Pro Val Arg Ile Pro Cys Leu Arg Gly Lys Phe Asn Leu Gln Leu Pro Ser Leu Asp Asp Gln Val Ile Pro Ala Arg Leu Pro Lys Thr Glu Val Ser Ala Glu Glu Pro Lys Glu Ala Thr Glu Val Lys Asp Gln Val Glu Thr Gln Gly Gln Glu Asp Asn Lys Arg Gly Pro Cys Ser Asn Gly Glu Ala Ala Ser Thr Ser Arg Pro Leu Glu Thr Gln Gly Asn Leu Thr Ser Ser Trp Tyr Asn Pro Arg Pro Leu Glu Gly Asn Val His Leu Lys Ser Leu Thr Glu Lys Asn Gln Thr Asp Lys Ala Gln Val His Ala Val <210> 83 <211> 155 <212> PRT
<213> Homo sapiens <400> 83 Met Gly Val Val Ser Leu Val Phe Leu Ile Ile Tyr Tyr Leu Asp Pro Ser Val Leu Ser Gly Val Ser Cys Phe Val Met Phe Leu Cys Leu Ala Asp Tyr Leu Val Pro Ile Leu Ala Pro Arg Ile Phe Gly Ser Asn Lys Trp Thr Thr Glu Gln Gln Gln Arg Phe His Glu Ile Cys Ser Asn Leu Val Lys Thr Arg Arg Arg Ala Val Gly Trp Trp Lys Arg Leu Phe Thr Leu Lys Glu Glu Lys Pro Lys Met Tyr Phe Met Thr Met Ile Val Ser Leu Ala Ala Val Ala Trp Val Gly Gln Gln Val His Asn Leu Leu Leu Thr Tyr Leu Ile Val Thr Ser Leu Leu Leu Leu Pro Gly Leu Asn Gln His Gly Ile Ile Leu Lys Tyr Ile Gly Met Ala Lys Arg Glu Ile Asn Lys Leu Leu Lys Gln Lys Glu Lys Lys Asn Glu <210> 84 <211> 155 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (92) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (105) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (125) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (130) <223> Xaa equals any of the naturally occurring L-amino acids <400> 84 Met Gln His Gln Leu His Leu Leu Ile Cys Trp Gly Lys Gly Ser Lys Ser Asn Thr Ser Cys Leu Gly Pro Val Leu Ser Cys Ser Asn Met Trp Ser Leu Ala Leu Leu Val Val Ala Gly Ser Met Gly Val Ala Tyr Ser Ser Val Val Met Tyr Val Leu Leu Trp Val Pro Leu Pro Leu Pro Ser His Phe Leu Pro Ser Gly Ala Pro Glu Ala Gln Pro Thr Thr Trp Ala Gln Ser Pro His Ser Val Cys Lys Cys Gly Thr Xaa Leu Gly Pro Ala Lys Pro Gln Gly Pro Ser Leu Pro Xaa Pro Pro Cys Leu Ile Met Leu Leu Ser Cys Arg Arg Gln Leu Gly Leu Ala Pro Ser Xaa Trp Leu Pro Gly Xaa Gly Ser His Gly Gly Glu Leu Arg Gly Cys Ser Gln Gly Trp Ala Pro Gly Ile Ala His Leu Asn Ile Cys Thr <210> 85 <211> 140 <212> PRT
<213> Homo Sapiens <400> 85 Ile Pro Leu Cys Ser Ile Phe Gly Ala Leu Ile Ala Val Cys Leu Ile Met Gly Leu Phe Asp Gly Cys Phe Ile Ser Ile Met Ala Pro Ile Ala Phe Glu Leu Val Gly Ala Gln Asp Val Ser Gln Ala Ile Gly Phe Leu Leu Gly Phe Met Ser Ile Pro Met Thr Val Gly Pro Pro Ile Ala Gly Leu Leu Arg Asp Lys Leu Gly Ser Tyr Asp Val Ala Phe Tyr Leu Ala Gly Val Pro Pro Leu Ile Gly Gly Ala Val Leu Cys Phe Ile Pro Trp Ile His Ser Lys Lys Gln Arg Glu Ile Ser Lys Thr Thr Gly Lys Glu Lys Met Glu Lys Met Leu Glu Asn Gln Asn Ser Leu Leu Ser Ser Ser Ser Gly Met Phe Lys Lys Glu Ser Asp Ser Ile Ile <210> 86 <211> 25 <212> PRT
<213> Homo Sapiens <400> 86 Met Ser Pro Ser Ile Arg Ile Leu Leu Val Leu Gln Gln Leu Gly Ser Leu Met Ala Pro Leu Pro Ser Ala His <210> 87 <211> 712 <212> PRT
<213> Homo Sapiens <400> 87 Met Gly Gln Gly Leu Lys Ala Trp Pro Arg Tyr Arg Val Val Gly Ser Ala Asp Ala Gly Gln Tyr Asn Leu Glu Ile Thr Asp Ala Glu Leu Ser Asp Asp Ala Ser Tyr Glu Cys Gln Ala Thr Glu Ala Ala Leu Arg Ser Arg Arg Ala Lys Leu Thr Val Leu Ile Pro Pro Glu Asp Thr Arg Ile Asp Gly Gly Pro Val Ile Leu Leu Gln Ala Gly Thr Pro His Asn Leu Thr Cys Arg Ala Phe Asn Ala Lys Pro Ala Ala Thr Ile Ile Trp Phe Arg Asp Gly Thr Gln Gln Glu Gly Ala Val Ala Ser Thr Glu Leu Leu Lys Asp Gly Lys Arg Glu Thr Thr Val Ser Gln Leu Leu Ile Asn Pro Thr Asp Leu Asp Ile Gly Arg Val Phe Thr Cys Arg Ser Met Asn Glu Ala Ile Pro Ser Gly Lys Glu Thr Ser Ile Glu Leu Asp Val His His Pro Pro Thr Val Thr Leu Ser Ile Glu Pro Gln Thr Val Gln Glu Gly Glu Arg Val Val Phe Thr Cys Gln Ala Thr Ala Asn Pro Glu Ile Leu Gly Tyr Arg Trp Ala Lys Gly Gly Phe Leu Ile Glu Asp Ala His Glu Ser Arg Tyr Glu Thr Asn Val Asp Tyr Ser Phe Phe Thr Glu Pro Val Ser Cys Glu Val His Asn Lys Val Gly Ser Thr Asn Val Ser Thr Leu Val Asn Val His Phe Ala Pro Arg Ile Val Val Asp Pro Lys Pro Thr Thr Thr Asp Ile Gly Ser Asp Val Thr Leu Thr Cys Val Trp Val Gly Asn Pro Pro Leu Thr Leu Thr Trp Thr Lys Lys Asp Ser Asn Met Gly Pro Arg Pro Pro Gly Ser Pro Pro Glu Ala Ala Leu Ser Ala Gln Val Leu Ser Asn Ser Asn Gln Leu Leu Leu Lys Ser Val Thr Gln Ala Asp Ala Gly Thr Tyr Thr Cys Arg Ala Ile Val Pro Arg Ile Gly Val Ala Glu Arg Glu Val Pro Leu Tyr Val Asn Gly Pro Pro Ile Ile Ser Ser Glu Ala Val Gln Tyr Ala Val Arg Gly Asp Gly Gly Lys Val Glu Cys Phe Ile Gly Ser Thr Pro Pro Pro Asp Arg Ile Ala Trp Ala Trp Lys Glu Asn Phe Leu Glu Val Gly Thr Leu Glu Arg Tyr Thr Val Glu Arg Thr Asn Ser Gly Ser Gly Val Leu Ser Thr Leu Thr Ile Asn Asn Val Met Glu Ala Asp Phe Gln Thr His Tyr Asn Cys Thr Ala Trp Asn Ser Phe Gly Pro Gly Thr Ala Ile Ile Gln Leu Glu Glu Arg Glu Val Leu Pro Val Gly Ile Ile Ala Gly Ala Thr Ile Gly Ala Ser Ile Leu Leu Ile Phe Phe Phe Ile Ala Leu Val Phe Phe Leu Tyr Arg Arg Arg Lys Gly Ser Arg Lys Asp Val Thr Leu Arg Lys Leu Asp Ile Lys Val Glu Thr Val Asn Arg Glu Pro Leu Thr Met His Ser Asp Arg Glu Asp Asp Thr Ala Ser Val Ser Thr Ala Thr Arg Val Met Lys Ala Ile Tyr Ser Ser Phe Lys Asp Asp Val Asp Leu Lys Gln Asp Leu Arg Cys Asp Thr Ile Asp Thr Arg Glu Glu Tyr Glu Met Lys Asp Pro Thr Asn Gly Tyr Tyr Asn Val Arg Ala His Glu Asp Arg Pro Ser Ser Arg Ala Val Leu Tyr Ala Asp Tyr Arg Ala Pro Gly Pro Ala Arg Phe Asp Gly Arg Pro Ser Ser Arg Leu Ser His Ser Ser Gly Tyr Ala Gln Leu Asn Thr Tyr Ser Arg Gly Pro Ala Ser Asp Tyr Gly Pro Glu Pro Thr Pro Pro Gly Pro Ala Ala Pro Ala Gly Thr Asp Thr Thr Ser Gln Leu Ser Tyr Glu Asn Tyr Glu Lys Phe Asn Ser His Pro Phe Pro Gly Ala Ala Gly Tyr Pro Thr Tyr Arg Leu Gly Tyr Pro Gln Ala Pro Pro Ser Gly Leu Glu Arg Thr Pro Tyr Glu Ala Tyr Asp Pro Ile Gly Lys Tyr Ala Thr Ala Thr Arg Phe Ser Tyr Thr Ser Gln His Ser Asp Tyr Gly Gln Arg Phe Gln Gln Arg Met Gln Thr His Val <210> 88 <211> 213 <212> PRT
<213> Homo sapiens <400> 88 Met Gln Ile Gln Val Ala Gly Leu Leu Gln Phe Ala Val Pro Leu Phe Ser Thr Ala Glu Glu Asp Leu Leu Ala Ile Gln Leu Leu Leu Asn Ser Ser Glu Ser Ser Leu His Gln Leu Thr Ala Met Val Asp Cys Arg Gly Leu His Lys Asp Tyr Leu Asp Ala Leu Ala Gly Ile Cys Tyr Asp Gly Leu Gln Gly Leu Leu Tyr Leu Gly Leu Phe Ser Phe Leu Ala Ala Leu Ala Phe Ser Thr Met Ile Cys Ala Gly Pro Arg Ala Trp Lys His Phe Thr Thr Arg Asn Arg Asp Tyr Asp Asp Ile Asp Asp Asp Asp Pro Phe Asn Pro Gln Ala Trp Arg Met Ala Ala His Ser Pro Pro Arg Gly Gln Leu His Ser Phe Cys Ser Tyr Ser Ser Gly Leu Gly Ser Gln Thr Ser Leu Gln Pro Pro Ala Gln Thr Ile Ser Asn Ala Pro Val Ser Glu Tyr Met Asn Gln Ala Met Leu Phe Gly Arg Asn Pro Arg Tyr Glu Asn Val Pro Leu Ile Gly Arg Ala Ser Pro Pro Pro Thr Tyr Ser Pro Ser Met Arg Ala Thr Tyr Leu Ser Val Ala Asp Glu His Leu Arg His Tyr Gly Asn Gln Phe Pro Ala <210> 89 <211> 346 <212> PRT
<213> Homo sapiens <400> 89 Met Val Glu Lys Glu Glu Ala Gly Gly Gly Ile Ser Glu Glu Glu Ala Ala Gln Tyr Asp Arg Gln Ile Arg Leu Trp Gly Leu Glu Ala Gln Lys Arg Leu Arg Ala Ser Arg Val Leu Leu Val Gly Leu Lys Gly Leu Gly Ala Glu Ile Ala Lys Asn Leu Ile Leu Ala Gly Val Lys Gly Leu Thr Met Leu Asp His Glu Gln Val Thr Pro Glu Asp Pro Gly Ala Gln Phe Leu Ile Arg Thr Gly Ser Val Gly Arg Asn Arg Ala Glu Ala Ser Leu Glu Arg Ala Gln Asn Leu Asn Pro Met Val Asp Val Lys Val Asp Thr Glu Asp Ile Glu Lys Lys Pro Glu Ser Phe Phe Thr Gln Phe Asp Ala Val Cys Leu Thr Cys Cys Ser Arg Asp Val Ile Val Lys Val Asp Gln Ile Cys His Lys Asn Ser Ile Lys Phe Phe Thr Gly Asp Val Phe Gly Tyr His Gly Tyr Thr Phe Ala Asn Leu Gly Glu His Glu Phe Val Glu 165 170 1'75 Glu Lys Thr Lys Val Ala Lys Val Ser Gln Gly Val Glu Asp Gly Pro Asp Thr Lys Arg Ala Lys Leu Asp Ser Ser Glu Thr Thr Met Val Lys Lys Lys Val Val Phe Cys Pro Val Lys Glu Ala Leu Glu Val Asp Trp Ser Ser Glu Lys Ala Lys Ala Ala Leu Lys Arg Thr Thr Ser Asp Tyr Phe Leu Leu Gln Val Leu Leu Lys Phe Arg Thr Asp Lys Gly Arg Asp Pro Ser Ser Asp Thr Tyr Glu Glu Asp Ser Glu Leu Leu Leu Gln Ile Arg Asn Asp Val Leu Asp Ser Leu Gly Ile Ser Pro Asp Leu Leu Pro Glu Asp Phe Val Arg Tyr Cys Phe Ser Glu Met Ala Pro Val Cys Ala Val Val Gly Gly Ile Leu Ala Gln Glu Ile Val Lys Ala Leu Ser Gln Arg Asp Pro Pro His Asn Asn Phe Phe Phe Phe Asp Gly Met Lys Gly Asn Gly Ile Val Glu Cys Leu Gly Pro Lys <210> 90 <211> 261 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (125) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (127) <223> Xaa equals any of the naturally occurring L-amino acids <400> 90 Met Ala Val Thr Ala Cys Gln Gly Leu Gly Phe Val Val Ser Leu Ile Gly Ile Ala Gly Ile Ile Ala Ala Thr Cys Met Asp Gln Trp Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala Val Phe Asn Tyr Gln Gly Leu Trp Arg Ser Cys Val Arg Glu Ser Ser Gly Phe Thr Glu Cys Arg Gly Tyr Phe Thr Leu Leu Gly Leu Pro Ala Met Leu Gln Ala Val Arg Ala Leu Met Ile Val Gly Ile Val Leu Gly Ala Ile Gly Leu Leu Val Ser Ile Phe Ala Leu Lys Cys Ile Arg Ile Gly Ser Met Glu Asp Ser Ala Lys Ala Asn Met Thr Leu Thr Ser Gly Ile Met Xaa Ile Xaa Ser Gly Leu Cys Ala Ile Ala Gly Val Ser Val Phe Ala Asn Met Leu Val Thr Asn Phe Trp Met Ser Thr Ala Asn Met Tyr Thr Gly Met Gly Gly Met Val Gln Thr Val Gln Thr Arg Tyr Thr Phe Gly Ala Ala Leu Phe Val Gly Trp Val Ala Gly Gly Leu Thr Leu Ile Gly Gly Val Met Met Cys Ile Ala Cys Arg Gly Leu Ala Pro Glu Glu Thr Asn Tyr Lys Ala Val Ser Tyr His Ala Ser Gly His Ser Val Ala Tyr Lys Pro Gly Gly Phe Lys Ala Ser Thr Gly Phe Gly Ser Asn Thr Lys Asn Lys Lys Arg Tyr Asp Gly Gly Ala Arg Thr Glu Asp Glu Val Gln Ser Tyr Pro Ser Lys His Asp Tyr Val <210> 91 <211> 360 <212> PRT

<213> Homo Sapiens <400> 91 Met Ala Ala Glu Trp Ala Ser Arg Phe Trp Leu Trp Ala Thr Leu Leu Ile Pro Ala Ala Ala Val Tyr Glu Asp Gln Val Gly Lys Phe Asp Trp Arg Gln Gln Tyr Val Gly Lys Val Lys Phe Ala Ser Leu Glu Phe Ser Pro Gly Ser Lys Lys Leu Val Val Ala Thr Glu Lys Asn Val Ile Ala Ala Leu Asn Ser Arg Thr Gly Glu Ile Leu Trp Arg His Val Asp Lys Gly Thr Ala Glu Gly Ala Val Asp Ala Met Leu Leu His Gly Gln Asp Val Ile Thr Val Ser Asn Gly Gly Arg Ile Met Arg Ser Trp Glu Thr Asn Ile Gly Gly Leu Asn Trp Glu Ile Thr Leu Asp Ser Gly Ser Phe Gln Ala Leu Gly Leu Val Gly Leu Gln Glu Ser Val Arg Tyr Ile Ala Val Leu Lys Lys Thr Thr Leu Ala Leu His His Leu Ser Ser Gly His Leu Lys Trp Val Glu His Leu Pro Glu Ser Asp Ser Ile His Tyr Gln Met Val Tyr Ser Tyr Gly Ser Gly Val Val Trp Ala Leu Gly Val Val Pro Phe Ser His Val Asn Ile Val Lys Phe Asn Val Glu Asp Gly Glu Ile Val Gln Gln Val Arg Val Ser Thr Pro Trp Leu Gln His Leu Ser Gly Ala Cys Gly Val Val Asp Glu Ala Val Leu Val Cys Pro Asp Pro Ser Ser Arg Ser Leu Gln Thr Leu Ala Leu Glu Thr Glu Trp Glu Leu Arg Gln Ile Pro Leu Gln Ser Leu Asp Leu Glu Phe Gly Ser Gly Phe Gln Pro Arg Val Leu Pro Thr Gln Pro Asn Pro Val Asp Ala Ser Arg Ala Gln Phe Phe Leu His Leu Ser Pro Ser His Tyr Ala Leu Leu Gln Tyr His Tyr Gly Thr Leu Ser Leu Leu Lys Asn Phe Pro Gln Thr Ala Leu Val Ser Phe Ala Thr Thr Gly Glu Lys Thr Val Ala Ala Val Met Ala Cys Arg Asn Glu Val Gln Lys Thr Ser Ser Ser Glu Asp Gly Ser Met Gly Glu Leu Phe Gly Glu Val <210> 92 <211> 485 <212> PRT
<213> Homo Sapiens <400> 92 Ala Arg Lys Pro Arg Ser Gln Ile Lys Asn Glu Ile Asn Ile Asp Thr Leu Ala Arg Asp Glu Phe Asn Leu Gln Lys Met Met Val Met Val Thr Ala Ser Gly Lys Leu Phe Gly Ile Glu Ser Ser Ser Gly Thr Ile Leu Trp Lys Gln Tyr Leu Pro Asn Val Lys Pro Asp Ser Ser Phe Lys Leu Met Val Gln Arg Thr Thr Ala His Phe Pro His Pro Pro Gln Cys Thr Leu Leu Val Lys Asp Lys Glu Ser Gly Met Ser Ser Leu Tyr Val Phe Asn Pro Ile Phe Gly Lys Trp Ser Gln Val Ala Pro Pro Val Leu Lys Arg Pro Ile Leu Gln Ser Leu Leu Leu Pro Val Met Asp Gln Asp Tyr Ala Lys Val Leu Leu Leu Ile Asp Asp Glu Tyr Lys Val Thr Ala Phe Pro Ala Thr Arg Asn Val Leu Arg Gln Leu His Glu Leu Ala Pro Ser Ile Phe Phe Tyr Leu Val Asp Ala Glu Gln Gly Arg Leu Cys Gly Tyr Arg Leu Arg Lys Asp Leu Thr Thr Glu Leu Ser Trp Glu Leu Thr Ile Pro Pro Glu Val Gln Arg Ile Val Lys Val Lys Gly Lys Arg Ser Ser Glu His Val His Ser Gln Gly Arg Val Met Gly Asp Arg Ser Val Leu Tyr Lys Ser Leu Asn Pro Asn Leu Leu Ala Val Val Thr Glu Ser Thr Asp Ala His His Glu Arg Thr Phe Ile Gly Ile Phe Leu Ile Asp Gly Val Thr Gly Arg Ile Ile His Ser Ser Val Gln Lys Lys Ala Lys Gly Pro Val His Ile Val His Ser Glu Asn Trp Val Val Tyr Gln Tyr Trp Asn Thr Lys Ala Arg Arg Asn Glu Phe Thr Val Leu Glu Leu Tyr Glu Gly Thr Glu Gln Tyr Asn Ala Thr Ala Phe Ser Ser Leu Asp Arg Pro Gln Leu Pro Gln Val Leu Gln Gln Ser Tyr Ile Phe Pro Ser Ser Ile Ser Ala Met Glu Ala Thr Ile Thr Glu Arg Gly Ile Thr Ser Arg His Leu Leu Ile Gly Leu Pro Ser Gly Ala Ile Leu Ser Leu Pro Lys Ala Leu Leu Asp Pro Arg Arg Pro Glu Ile Pro Thr Glu Gln Ser Arg Glu Glu Asn Leu Ile Pro Tyr Ser Pro Asp Val Gln Ile His Ala Glu Arg Phe Ile Asn Tyr Asn Gln Thr Val Ser Arg Met Arg Gly Ile Tyr Thr Ala Pro Ser Gly Leu Glu Ser Thr Cys Leu Val Val Ala Tyr Gly Leu Asp Ile Tyr Gln Thr Arg Val Tyr Pro Ser Lys Gln Phe Asp Val Leu Lys Asp Asp Tyr Asp Tyr Val Leu Ile Ser Ser Val Leu Phe Gly Leu Val Phe Ala Thr Met Ile Thr Lys Arg Leu Ala Gln Val Lys Leu Leu Asn Arg Ala Trp Arg <210> 93 <211> 149 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (140) <223> Xaa equals any of the naturally occurring L-amino acids <400> 93 Met Val Leu Leu Trp Ala Ser Val Leu Phe Pro Ala Pro Glu Asp Trp Ala Glu Leu Gln Gly Ala Val Tyr Arg Leu Leu Val Val Leu Leu Cys Cys Leu Ala Thr Arg Lys Leu Pro His Phe Leu His Pro Gln Arg Asn Leu Leu Gln Gly Ser Gly Leu Asp Leu Gly Ala Ile Tyr Gln Arg Val Glu Gly Phe Ala Ser Gln Pro Glu Ala Ala Leu Arg Ile His Ala Thr His Leu Gly Arg Ser Pro Pro Pro Arg Ile Gly Ser Gly Leu Lys Ala Leu Leu Gln Leu Pro Ala Ser Asp Pro Thr Tyr Trp Ala Thr Ala Tyr Phe Asp Val Leu Leu Asp Lys Phe Gln Val Phe Asn Ile Gln Asp Lys Asp Arg Ile Ser Ala Met Gln Ser Ile Phe Gln Xaa Thr Arg Thr Leu Gly Gly Glu Glu Ser <210> 94 <211> 209 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (136) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (161) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE

<222> (169) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (197) <223> Xaa equals any of the naturally occurring L-amino acids <400> 94 Met Ala Met Gly Leu Phe Arg Val Cys Leu Val Val Val Thr Ala Ile Ile Asn His Pro Leu Leu Phe Pro Arg Glu Asn Ala Thr Val Pro Glu Asn Glu Glu Glu Ile Ile Arg Lys Met Gln Ala His Gln Glu Lys Leu Gln Leu Glu Gln Leu Arg Leu Glu Glu Glu Val Ala Arg Leu Ala Ala Glu Lys Glu Ala Leu Glu Gln Val Ala Glu Glu Gly Arg Gln Gln Asn Glu Thr Arg Val Ala Trp Asp Leu Trp Ser Thr Leu Cys Met Ile Leu Phe Leu Met Ile Glu Val Trp Arg Gln Asp His Gln Glu Gly Pro Ser Pro Glu Cys Leu Gly Gly Glu Glu Asp Glu Leu Pro Gly Trp Gly Ala Pro Pro Cys Arg Ala Ser Pro Xaa Pro Thr Arg His Ala Cys His Phe Tyr Glu Arg Cys Ile Arg Gly Ala Thr Ala Asp Ala Ala Arg Thr Arg Xaa Phe Leu Glu Gly Phe Val Asp Xaa Leu Leu Glu Ala Leu Arg Ser Leu Cys Asn Arg Asp Thr Asp Met Glu Val Glu Asp Phe Ile Gly Val Asp Ser Met Tyr Xaa Asn Trp Gln Val Asp Arg Pro Leu Leu Cys His Leu <210> 95 <211> 107 <212> PRT
<213> Homo sapiens <400> 95 Met Leu Val Leu Leu Leu Asp Phe Leu Gly Leu Val His Leu Gly Gln Leu Leu Ile Phe His Ile Tyr Leu Lys Ala Lys Lys Met Thr Thr Phe Glu Tyr Leu Ile Asn Asn Arg Lys Glu Glu Ser Ser Lys His Gln Ala Val Arg Lys Asp Pro Tyr Val Gln Met Asp Lys Gly Val Leu Gln Gln Gly Ala Gly Ala Leu Gly Ser Ser Ala Gln Gly Val Lys Ala Lys Ser Ser Leu Leu Ile His Lys His Leu Cys His Phe Cys Thr Ser Val Asn Gln Asp Gly Asp Ser Thr Ala Arg Val His Leu <210> 96 <211> 6 <212> PRT
<213> Homo Sapiens <400> 96 Val Phe Gln Ile Tyr Leu <210> 97 <211> 6 <212> PRT
<213> Homo Sapiens <400> 97 Val Phe Gln Ile Tyr Leu <210> 98 <211> 357 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (140) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (325) <223> Xaa equals any of the naturally occurring L-amino acids <220>

<221> SITE
<222> (329) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (335) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (338) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (339) <223> Xaa equals any of the naturally occurring L-amino acids <400> 98 Met Gly Met Gly Arg Gly Ala Gly Arg Ser Ala Leu Gly Phe Trp Pro Thr Leu Ala Phe Leu Leu Cys Ser Phe Pro Ala Ala Thr Ser Pro Cys Lys Ile Leu Lys Cys Asn Ser Glu Phe Trp Ser Ala Thr Ser Gly Ser His Ala Pro Ala Ser Asp Asp Thr Pro Glu Phe Cys Ala Ala Leu Arg Ser Tyr Ala Leu Cys Thr Arg Arg Thr Ala Arg Thr Cys Arg Gly Asp Leu Ala Tyr His Ser Ala Val His Gly Ile Glu Asp Leu Met Ser Gln His Asn Cys Ser Lys Asp Gly Pro Thr Ser Gln Pro Arg Leu Arg Thr Leu Pro Pro Ala Gly Asp Ser Gln Glu Arg Ser Asp Ser Pro Glu Ile Cys His Tyr Glu Lys Ser Phe His Lys His Ser Xaa Thr Pro Asn Tyr Thr His Cys Gly Leu Phe Gly Asp Pro His Leu Arg Thr Phe Thr Asp Arg Phe Gln Thr Cys Lys Val Gln Gly Ala Trp Pro Leu Ile Asp Asn Asn Tyr Leu Asn Val Gln Val Thr Asn Thr Pro Val Leu Pro Gly Ser Ala Ala Thr Ala Thr Ser Lys Leu Thr Ile Ile Phe Lys Asn Phe Gln Glu Cys Val Asp Gln Lys Val Tyr Gln Ala Glu Met Asp Glu Leu Pro Ala Ala Phe Val Asp Gly Ser Lys Asn Gly Gly Asp Lys His Gly Ala Asn Ser Leu Lys Ile Thr Glu Lys Val Ser Gly Gln His Val Glu Ile Gln Ala Lys Tyr Ile Gly Thr Thr Ile Val Val Arg Gln Val Gly Arg Tyr Leu Thr Phe Ala Val Arg Met Pro Glu Glu Val Val Asn Ala Val Glu Asp Trp Asp Ser Gln Gly Leu Tyr Leu Cys Leu Arg Gly Cys Pro Leu Asn Gln Gln Ile Asp Phe Gln Ala Phe His Thr Asn Ala Glu Gly Thr Gly Ala Arg Xaa Leu Ala Ala Xaa Ser Leu Asp Pro Gln Xaa Pro Arg Xaa Xaa His Thr Arg Gln Ala Val Ala Lys Cys Lys Glu Lys Leu Pro Val Glu Asp Leu <210> 99 <211> 179 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (91) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (140) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (169) <223> Xaa equals any of the naturally occurring L-amino acids <400> 99 Met Gly Met Gly Arg Gly Ala Gly Arg Ser Ala Leu Gly Phe Trp Pro Thr Leu Ala Phe Leu Leu Cys Ser Phe Pro Ala Ala Thr Ser Pro Cys Lys Ile Leu Lys Cys Asn Ser Glu Phe Trp Ser Ala Thr Ser Gly Ser His Ala Pro Ala Ser Asp Asp Thr Pro Glu Phe Cys Ala Ala Leu Arg Ser Tyr Ala Leu Cys Thr Arg Arg Thr Ala Arg Thr Cys Arg Gly Asp Leu Ala Tyr His Ser Ala Val His Gly Ile Xaa Asp Leu Met Ser Gln His Asn Cys Ser Lys Asp Gly Pro Thr Ser Gln Pro Arg Leu Arg Thr Leu Pro Pro Ala Glu Thr Ala Arg Ser Ala Arg Thr Ala Pro Arg Ser Ala Ile Thr Arg Arg Ala Phe Thr Ser Thr Arg Xaa Pro Pro Thr Thr Arg Thr Val Ala Ser Ser Gly Thr His Thr Phe Arg Thr Phe Thr Asp Arg Phe Gln Thr Cys Lys Val Gln Xaa Arg Leu Ala Ala His Arg Gln Leu Ile Thr <210> 100 <211> 213 <212> PRT
<213> Homo sapiens <400> 100 Leu Leu Phe Leu Ser Leu Leu Gln Met Gln Glu Leu Leu Gly Arg Gly Ala Trp Ala Pro Gly Cys Gly Arg Arg Pro Ser Gly Trp Gly Gln Leu Ala Cys Pro Asp Pro Leu Leu Pro Pro His Asn Pro Lys Ser Pro Gln Pro Gly Pro Ser Thr Ser Gly Val Trp Gly Glu Glu Gln Gly Leu Arg Thr Leu Ser Ser Glu His Pro Trp Gln Gly Leu Gln Pro Leu Ile Ser Ser Leu Lys Pro Cys Gly His Thr Ala Arg Arg Asp Leu Pro Leu Ala Pro Ala Ser Phe Gln Pro Arg Val Leu Ile Gln Gly Pro Arg Thr Val Pro Pro Val Leu Leu Cys Pro Gln His Lys Ala Arg Leu His Ser Gln Lys Cys Ser Gln Ala Leu Glu Gly Asp Pro Ala Ser Ser Pro Thr Ala Pro His Pro Thr His Pro Ser Ala Ala Pro Leu Leu Phe Pro Arg Asp Leu Ser Tyr Thr Gly Gln Glu Ala Ala Glu Arg Val Ser Pro Pro Pro Ser Lys Arg Ser Cys Ser Leu Cys Gln Asn Arg Val Trp Ala Gly Gly Arg Ala Leu Gly Ala Arg Pro Leu Pro Leu Pro Ala Gly Phe Ser Trp Ser Leu Cys Trp Lys <210> 101 <211> 94 <212> PRT
<213> Homo sapiens <400> 101 Met Leu Leu Gly Phe Leu Val Leu Ile Pro Trp Gly Ser Leu Ile Leu Gly Ser Ser Asp Leu Asp Pro Ser Ser Leu Pro Leu Gly Thr Arg Gly His Gly Trp Arg Trp Pro Pro Leu Ser Pro Val Gln Ile Leu Tyr Pro Leu Ala Gly Asp Pro His Ala Ala Val Ser Cys Ser Cys Cys Gly Glu Thr Glu Leu Arg Ala Leu Leu Thr Gly Ser Leu Pro Met Glu Ala Phe Ser Gly Leu His Ser Ile Glu Tyr Ser Ser Arg Thr Ala Cys <210> 102 <211> 135 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (125) <223> Xaa equals any of the naturally occurring L-amino acids <400> 102 Met Leu His Leu Lys Ala Leu Gly Arg Gly Leu Leu Gln Ala Cys Leu Leu Ala Thr Gly Gly Ser Met Ala Cys Asp Ser Ile Thr Pro Ile Phe Thr Gln His Ser Pro Arg Val Pro Leu Ser Pro Ser Phe Pro Leu Leu Glri Glu Thr Gly Ala Phe Ala Pro Ser Pro Arg Leu Glu Gly Ser Gly Met Ile Leu Ala His Cys Cys Leu Glu Leu Leu Cys Ser Ser Asp Pro Pro Ala Ser Ala Ile Gln Asn Cys Trp Asp Tyr Arg Cys Glu Pro Arg His Leu Ala Ser Ile Ser Ser Phe Tyr Lys Asp Ile Gly Arg Ile Gly Leu Gly Pro Val Leu Leu Gln Tyr Asn Leu Ile Leu Xaa Asn Tyr Ile Cys His Asn Ser Val Ser Lys <210> 103 <211> 92 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (68) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (86) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (90) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (92) <223> Xaa equals any of the naturally occurring L-amino acids <400> 103 Met Leu His Leu Lys Ala Leu Gly Arg Gly Leu Leu Gln Ala Cys Leu Leu Ala Thr Gly Gly Ser Met Ala Cys Asp Ser Ile Thr Pro Ile Phe Thr Gln His Ser Pro Arg Val Pro Leu Ser Pro Ser Phe Pro Leu Leu Gln Glu Thr Gly Ala Phe Ala Pro Ser Pro Arg Leu Glu Gly Ser Gly Met Ile Leu Xaa His Cys Cys Leu Glu Leu Leu Cys Ser Ser Asp Pro Pro Ala Ser Ala Ile Xaa Asn Cys Trp Xaa Tyr Xaa <210> 104 <211> 75 <212> PRT
<213> Homo Sapiens <400> 104 Met Leu Asn Pro Leu Arg Gln Leu Phe Lys Leu Met Ala Ser Leu Phe Leu Ser Val Phe Thr Leu Gly Leu Pro Phe Ala Leu Phe Gln Tyr Tyr Ala Tyr Thr Gln Phe Cys Leu Pro Gly Ser Ala Arg Pro Ile Pro Glu Pro Leu Val Gln Leu Ala Val Asp Lys Gly Tyr Arg Ile Ala Glu Gly Asn Glu Pro Leu Gly Ala Ser Gly Met Phe His <210> 105 <211> 28 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (3) <223> Xaa equals any of the naturally occurring L-amino acids <400> 105 Glu Cys Xaa Arg Lys Pro Thr Pro Arg Ala Glu Phe Leu Gln Pro Gly Gly Ser Thr Ser Ser Arg Ala Ala Ala Thr Ala Val <210> 106 <211> 105 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (69) <223> Xaa equals any of the naturally occurring L-amino acids <400> 106 Met Phe Ala Leu Ala Trp Lys Val Ile Phe Ser Val Met Leu Gln Asn Pro Ile Arg Tyr Pro Ser Val Leu Gly Ile Lys Ser Ser Leu Leu Ser Ser Leu Val Leu Val Met Val Trp Gly Asn Glu Lys Ser Gly Pro Cys Pro Thr Pro Lys Ser Arg Lys Gly Arg Arg Ser Cys Pro Ala Gln Val Gly Arg Gly Glu Xaa Gly Ser Tyr Trp Asp Pro Glu Phe Arg Leu Ser Arg Lys Ser Asn Gln Gly Leu Arg Arg Asp Tyr Leu Ser Leu Tyr His Phe Asn Leu His Phe Arg Asp Thr Phe <210> 107 <211> 92 <212> PRT
<213> Homo sapiens <400> 107 Met Thr Ser Leu Leu Glu Gly Arg Met Val Leu Cys Val Ser Cys Leu Leu Leu Pro Leu Leu Leu Leu Leu Lys His Phe Asn Gly Leu Met Thr Pro Tyr Leu Ala His Asn Val Tyr Cys Pro Ile Glu Tyr Ile Ser Phe Phe Pro Phe His Glu Lys Asn Ile Glu Tyr Ile Ser Ile Trp Phe Ile Phe Asp Ser Phe Lys Phe Ile Tyr Ser Arg Leu Leu Cys Ile Ser Gln Ile Tyr Val Leu Tyr Arg Ala Tyr Thr Leu Pro His <210> 108 <211> 92 <212> PRT
<213> Homo.sapiens <400> 108 Met Thr Ser Leu Leu Glu Gly Arg Met Val Leu Cys Val Ser Cys Leu Leu Leu Pro Leu Leu Leu Leu Leu Lys His Phe Asn Gly Leu Met Thr Pro Tyr Leu Ala His Asn Val Tyr Cys Pro Ile Glu Tyr Ile Ser Phe Phe Pro Phe His Glu Lys Asn Ile Glu Tyr Ile Ser Ile Trp Phe Ile Phe Asp Ser Phe Lys Phe Ile Tyr Ser Arg Leu Leu Cys Ile Ser Gln Ile Tyr Val Leu Tyr Arg Ala Tyr Thr Leu Pro His <210> 109 <211> 182 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (104) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (147) <223> Xaa equals any of the riaturally occurring L-amino acids <220>
<221> SITE
<222> (151) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (176) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (179) <223> Xaa equals any of the naturally occurring L-amino acids <400> 109 Met Leu Trp Phe Ser Gly Val Gly Ala Leu Ala Glu Arg Tyr Cys Arg Arg Ser Pro Gly Ile Thr Cys Cys Val Leu Leu Leu Leu Asn Cys Ser Gly Val Pro Met Ser Leu Ala Ser Ser Phe Leu Thr Gly Ser Val Ala Lys Cys Glu Asn Glu Gly Glu Val Leu~Gln Ile Pro Phe Ile Thr Asp Asn Pro Cys Ile Met'Cys Val Cys Leu Asn Lys Glu Val Thr Cys Lys Arg Glu Lys Cys Pro Val Leu Ser Arg Asp Cys Ala Leu Ala Ile Lys Gln Arg Gly Ala Cys Cys Glu Xaa Cys Lys Gly Cys Thr Tyr Glu Gly Asn Thr Tyr Asn Ser Ser Phe Lys Trp Gln Ser Pro Ala Glu Pro Cys Val Leu Arg Gln Cys Gln Glu Gly Val Val Thr Glu Ser Gly Val Arg Cys Val Xaa His Cys Lys Xaa Pro Leu Glu His Leu Gly Met Cys Cys Pro Thr Cys Pro Gly Cys Val Phe Glu Gly Val Gln Tyr Gln Glu Xaa Glu Glu Xaa Gln Pro Glu <210> 110 <211> 47 <212> PRT
<213> Homo Sapiens <400> 110 Ser Ser Ser Leu Leu Ile Ile Tyr Val Cys Met Met Asp Val Thr Ile Tyr Met Ser Cys Val Glu Ile Lys Gly Cys Leu Asp Ala Met Leu Ile Leu Leu Ser Met Arg Lys Tyr Leu Lys Lys Leu Leu His Asn Ile <210> 111 <211> 96 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (81) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (83) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (84) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (96) <223> Xaa equals any of the naturally occurring L-amino acids <400> 111 Met Lys Val Ala Val Ser Pro Ala Val Gly Pro Gly Pro Trp Gly Ser Gly Val Gly Gly Gly Gly Thr Val Arg Leu Leu Leu Ile Leu Ser Gly Cys Leu Val Tyr Gly Thr Ala Glu Thr Asp Val Asn Val Val Met Leu Gln Glu Ser Gln Val Cys Glu Lys Arg Ala Ser Gln Gln Phe Cys Tyr Thr Asn Val Leu Ile Pro Lys Trp His Asp Ile Trp Thr Arg Ile Gln Xaa Arg Xaa Xaa Ser Ser Arg Leu Val Arg Val Thr Gln Val Glu Xaa <210> 112 <211> 273 <212> PRT
<213> Homo Sapiens <400> 112 Met Leu Phe Phe Cys Gly Asp Leu Leu Ser Arg Ser Gln Ile Phe Tyr Tyr Ser Thr Gly Met Thr Val Gly Ile Val Ala Ser Leu Leu Ile Ile Ile Phe Ile Leu Ser Lys Phe Met Pro Lys Lys Ser Pro Ile Tyr Val Ile Leu Val Gly Gly Trp Ser Phe Ser Leu Tyr Leu Ile Gln Leu Val Phe Lys Asn Leu Gln Glu Ile Trp Arg Cys Tyr Trp Gln Tyr Leu Leu Ser Tyr Val Leu Thr Val Gly Phe Met Ser Phe Ala Val Cys Tyr Lys Tyr Gly Pro Leu Glu Asn Glu Arg Ser Ile Asn Leu Leu Thr Trp Thr Leu Gln Leu Met Gly Leu Cys Phe Met Tyr Ser Gly Ile Gln Ile Pro His Ile Ala Leu Ala Ile Ile Ile Ile Ala Leu Cys Thr Lys Asn Leu Glu His Pro Ile Gln Trp Leu Tyr Ile Thr Cys Arg Lys Val Cys Lys Gly Ala Glu Lys Pro Val Pro Pro Arg Leu Leu Thr Glu Glu Glu Tyr Arg Ile Gln Gly Glu Val Glu Thr Arg Lys Ala Leu Glu Glu Leu Arg Glu Phe Cys Asn Ser Pro Asp Cys Ser Ala Trp Lys Thr Val Ser Arg Ile Gln Ser Pro Lys Arg Phe Ala Asp Phe Val Glu Gly Ser Ser His Leu Thr Pro Asn Glu Val Ser Val His Glu Gln Glu Tyr Gly Leu Gly Ser Ile Ile Ala Gln Asp Glu Ile Tyr Glu Glu Ala Ser Ser Glu Glu Glu Asp Ser Tyr Ser Arg Cys Pro Ala Ile Thr Gln Asn Asn Phe Leu Thr <210> 113 <211> 265 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (22) <223> Xaa equals any of the naturally occurring L-amino acids <400> 113 Pro Lys Leu Glu Leu His Arg Gly Gly Gly Arg Ser Arg Thr Ser Gly Ser Pro Gly Leu Gln Xaa Phe Gly Thr Arg Arg Thr Arg Gly Arg Ser Gly Arg Ala Gln Gly Arg Leu Lys Arg Pro Gly Lys Leu Ala Cys Arg Lys Phe Pro Gly Arg Arg Gln Arg Val Val Pro Glu Leu Thr Asp Val Leu Met Asn Glu Ile Leu His Gly Ala Asp Gly Thr Ser Ile Lys Cys Gly Ile Ile Gly Glu Ile Gly Cys Ser Trp Pro Leu Thr Glu Ser Glu Arg Lys Val Leu Gln Ala Thr Ala His Ala Gln Ala Gln Leu Gly Cys Pro Val Ile Ile His Pro Gly Arg Ser Ser Arg Ala Pro Phe Gln Ile Ile Arg Ile Leu Gln Glu Ala Gly Ala Asp Ile Ser Lys Thr Val Met Ser His Leu Asp Arg Thr Ile Leu Asp Lys Lys Glu Leu Leu Glu Phe Ala Gln Leu Gly Cys Tyr Leu Glu Tyr Asp Leu Phe Gly Thr Glu Leu Leu His Tyr Gln Leu Gly Pro Asp Ile Asp Met Pro Asp Asp Asn Lys Arg Ile Arg Arg Val Arg Leu Leu Val Glu Glu Gly Cys Glu Asp Arg Ile Leu Val Ala His Asp Ile His Thr Lys Thr Arg Leu Met Lys Tyr Gly Gly His Gly Tyr Ser His Ile Leu Thr Asn Val Val Pro Lys Met Leu Leu Arg Gly Ile Thr Glu Asn Val Leu Asp Lys Ile Leu Ile Glu Asn Pro Lys Gln Trp Leu Thr Phe Lys <210> 114 <211> 91 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (78) <223> Xaa equals any of the naturally occurring L-amino acids <400> 114 Met His Leu Gly Leu Val Ser Leu Ile Leu Phe Cys Gln Ala Leu Glu Val Asp Ile Ser Leu Gln Gly Pro Gly Ile Val Pro Gly Arg Ser Glu Val Ser Leu Ser Leu Gln Gly Pro Arg Gly Gly Gly Cys Phe Pro Ile Ala Thr Gly Ala Pro Phe Ile Val Leu Leu Pro Leu Gly Leu Tyr Leu Val Phe His Leu Cys Cys Phe Phe Gly Leu Phe Cys Ala Xaa Leu Arg Leu Arg Glu Pro Gly Trp Asp His Leu Ile Ile <210> 115 <211> 81 .
<212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (56) <223> Xaa equals any of the naturally occurring L-amino acids <400> 115 Gly Leu His Phe Asn Ile Arg Val Asp His Gly Met Leu Trp Ala Pro Val Leu Tyr Lys Asp Val Gly Gln Glu Leu Pro Val Val Ser Thr Ala Pro Ser His Ile Ala Leu Leu Met Glu Pro Phe Thr Pro Asp Val Leu Ser Arg Leu Met Gly Arg Ile Xaa Val Cys Lys Asp Tyr Val Ile Asp Gln Leu Trp Ser Val Leu Lys Glu Ile Cys Gln Trp Ile Ile Pro Tyr Gly <210> 116 <211> 234 <212> PRT
<213> Homo Sapiens <400> 116 Pro Thr Arg Pro Pro Thr Arg Pro Val Arg Val Ser Val Gly Gly Leu Val Gly Glu Val Ala Cys Ala Cys Arg Asp Cys Ile Pro Glu Thr Met Ala Glu Gly Asp Asn Arg Ser Thr Asn Leu Leu Ala Ala Glu Thr Ala Ser Leu Glu Glu Gln Leu Gln Gly Trp Gly Glu Val Met Leu Met Ala Asp Lys Val Leu Arg Trp Glu Arg Ala Trp Phe Pro Pro Ala Ile Met Gly Val Val Ser Leu Val Phe Leu Ile Ile Tyr Tyr Leu Asp Pro Ser Val Leu Ser Gly Val Ser Cys Phe Val Met Phe Leu Cys Leu Ala Asp Tyr Leu Val Pro Ile Leu Ala Pro Arg Ile Phe Gly Ser Asn Lys Trp Thr Thr Glu Gln Gln Gln Arg Phe His Glu Ile Cys Ser Asn Leu Val 130 135 ' 140 Lys Thr Arg Arg Arg Ala Val Gly Trp Trp Lys Arg Leu Phe Thr Leu Lys Glu Glu Lys Pro Lys Met Tyr Phe Met Thr Met Ile Val Ser Leu 165 170 ' 175 Ala Ala Val Ala Trp Val Gly Gln Gln Val His Asn Leu Leu Leu Thr Tyr Leu Ile Val Thr Ser Leu Leu Leu Leu Pro Gly Leu Asn Gln His Gly Ile Ile Leu Lys Tyr Ile Gly Met Ala Lys Arg Glu Ile Asn Lys Leu Leu Lys Gln Lys Glu Lys Lys Asn Glu <210> 117 <211> 42 <212> PRT
<213> Homo Sapiens <400> 117 ' Tyr Thr Phe Gln Cys Leu Ser Gln Thr Cys Ser Tyr Asp Ile Lys Cys Tyr Phe Leu Val Ala Lys Ile Ile Leu Asp Ser Val Ile Lys Val Tyr Trp Asn Leu Asn Phe Lys Met Ser Pro Asp <210> 118 <211> 25 <212> PRT
<213> Homo sapiens <400> 118 Met Ser Pro Ser Ile Arg Ile Leu Leu Val Leu Gln Gln Leu Gly Ser Leu Met Ala Pro Leu Pro Ser Ala His <210> 119 <211> 156 <212> PRT
<213> Homo sapiens <400> 119 Met Gln Ile Gln Val Ala Gly Leu Leu Gln Phe Ala Val Pro Leu Phe Ser Thr Ala Glu Glu Asp Leu Leu Ala Ile Gln Leu Leu Leu Asn Ser Ser Glu Ser Ser Leu His Gln Leu Thr Ala Met Val Asp Cys Arg Gly Leu His Lys Asp Tyr Leu Asp Ala Leu Ala Gly Ile Cys Tyr Asp Gly Leu Gln Gly Leu Leu Tyr Leu Gly Leu Phe Ser Phe Leu Ala Ala Leu Ala Phe Ser Thr Met Ile Cys Ala Gly Pro Arg Ala Trp Lys His Phe Thr Thr Arg Asn Arg Asp Tyr Asp Asp Ile Asp Asp Asp Asp Pro Phe Asn Pro Gln Ala Trp Arg Met Ala Ala His Ser Pro Pro Arg Gly Gln Leu His Ser Phe Cys Ser Tyr Ser Ser Gly Leu Gly Ser Gln Thr Ser Leu Gln Pro Pro Ala Gln Thr Ile Ser Asn Ala Pro <210> 120 <211> 42 <212> PRT
<213> Homo sapiens <400> 120 Met Val Pro Ser Val Thr Leu Ile Leu His Cys Pro Gly Phe Ser Thr Glu Ser His Met Cys Gly Lys Pro Leu Ser Pro Arg Pro Thr Arg Thr Val Gly Arg Pro Val Ser Asn Ile Pro Val <210> 121 <211> 89 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (17) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (47) <223> Xaa equals any of the naturally occurring L-amino acids <400> 121 Val Gln Gly Val Val Gln Ala Leu Lys Thr Asp His Ala Phe Cys Pro Xaa Leu Gln Gly Thr Glu Ser Ile Arg Leu Arg Ile Leu Glu Phe Glu Leu Asn Gln Val Arg Ser Val Ser Gln Glu Leu Pro Pro Gly Xaa Pro Glu Ser Pro Gln Thr Asp Gly Gln Pro Pro Arg Ala Trp Pro Gln Leu Gly Met Pro Ser Asn Pro Thr Cys Phe Ser Phe Leu Pro Gly Tyr Ser 65 , 70 75 80 Gly Leu Arg Ser Ser Ala Leu Asn Phe <210> 122 <211> 370 <212> PRT
<213> Homo Sapiens <400> 122 Leu Gly Gly Ala Arg Val Arg Arg Ala Val Gly Leu Ser Gly Thr Gly Ala Glu Ala Gly Arg Ala Gly Ala Met Val Glu Lys Glu Glu Ala Gly Gly Gly Ile Ser Glu Glu Glu Ala Ala Gln Tyr Asp Arg Gln Ile Arg Leu Trp Gly Leu Glu Ala Gln Lys Arg Leu Arg Ala Ser Arg Val Leu Leu Val Gly Leu Lys Gly Leu Gly Ala Glu Ile Ala Lys Asn Leu Ile Leu Ala Gly Val Lys Gly Leu Thr Met Leu Asp His Glu Gln Val Thr Pro Glu Asp Pro Gly Ala Gln Phe Leu Ile Arg Thr Gly Ser Val Gly Arg Asn Arg Ala Glu Ala Ser Leu Glu Arg Ala Gln Asn Leu Asn Pro Met Val Asp Val Lys Val Asp Thr Glu Asp Ile Glu Lys Lys Pro Glu Ser Phe Phe Thr Gln Phe Asp Ala Val Cys Leu Thr Cys Cys Ser Arg Asp Val Ile Val Lys Val Asp Gln Ile Cys His Lys Asn Ser Ile Lys Phe Phe Thr Gly Asp Val Phe Gly Tyr His Gly Tyr Thr Phe Ala Asn Leu Gly Glu His Glu Phe Val Glu Glu Lys Thr Lys Val Ala Lys Val Ser Gln Gly Val Glu Asp Gly Pro Asp Thr Lys Arg Ala Lys Leu Asp Ser Ser Glu Thr Thr Met Val Lys Lys Lys Val Val Phe Cys Pro Val Lys Glu Ala Leu Glu Val Asp Trp Ser Ser Glu Lys Ala Lys Ala Ala Leu Lys Arg Thr Thr Ser Asp Tyr Phe Leu Leu Gln Val Leu Leu Lys Phe Arg Thr Asp Lys Gly Arg Asp Pro Ser Ser Asp Thr Tyr Glu Glu Asp Ser Glu Leu Leu Leu Gln Ile Arg Asn Asp Val Leu Asp Ser Leu Gly Ile Ser Pro Asp Leu Leu Pro Glu Asp Phe Val Arg Tyr Cys Phe Ser Glu Met Ala Pro Val Cys Ala Val Val Gly Gly Ile Leu Ala Gln Glu Ile Val Lys Ala Leu Ser Gln Arg Asp Pro Pro His Asn Asn Phe Phe Phe Phe Asp Gly Met Lys Gly Asn Gly Ile Val Glu Cys Leu Gly Pro Lys <210> 123 <211> 276 <212> PRT
<213> Homo Sapiens <400> 123 Gly Thr Ser Leu Ser Cys Gly Ser Val Ser Thr Leu Cys Ala Thr Met Ala Val Thr Ala Cys Gln Gly Leu Gly Phe Val Val Ser Leu Ile Gly Ile Ala Gly Ile Ile Ala Ala Thr Cys Met Asp Gln Trp Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala Val Phe Asn Tyr Gln Gly Leu Trp Arg Ser Cys Val Arg Glu Ser Ser Gly Phe Thr Glu Cys Arg Gly Tyr Phe Thr Leu Leu Gly Leu Pro Ala Met Leu Gln Ala Val Arg Ala Leu Met Ile Val Gly Ile Val Leu Gly Ala Ile Gly Leu Leu Val Ser Ile Phe Ala Leu Lys Cys Ile Arg Ile Gly Ser Met Glu Asp Ser Ala Lys Ala Asn Met Thr Leu Thr Ser Gly Ile Met Phe Ile Val Ser Gly Leu Cys Ala Ile Ala Gly Val Ser Val Phe Ala Asn Met Leu Val Thr Asn Phe Trp Met Ser Thr Ala Asn Met Tyr Thr Gly Met Gly Gly Met Val Gln Thr Val Gln Thr Arg Tyr Thr Phe Gly Ala Ala Leu Phe Val 180 . 185 190 Gly Trp Val Ala Gly Gly Leu Thr Leu Ile Gly Gly Val Met Met Cys Ile Ala Cys Arg Gly Leu Ala Pro Glu Glu Thr Asn Tyr Lys Ala Val Ser Tyr His Ala Ser Gly His Ser Val Ala Tyr Lys Pro Gly Gly Phe Lys Ala Ser Thr Gly Phe Gly Ser Asn Thr Lys Asn Lys Lys Arg Tyr Asp Gly Gly Ala Arg Thr Glu Asp Glu Val Gln Ser Tyr Pro Ser Lys His Asp Tyr Val <210> 124 <211> 75 <212> PRT
<213> Homo sapiens <400> 124 Met Arg Gly Ile Tyr Thr Ala Pro Ser Gly Leu Glu Ser Thr Cys Leu 1 ~ 5 10 15 Val Val Ala Tyr Gly Leu Asp Ile Tyr Gln Thr Arg Val Tyr Pro Ser Lys Gln Phe Asp Val Leu Lys Asp Asp Tyr Asp Tyr Val Leu Ile Ser Ser Val Leu Phe Gly Leu Val Phe Ala Thr Met Ile Thr Lys Arg Leu Ala Gln Val Lys Leu Leu Asn Arg Ala Trp Arg <210> 125 <211> 171 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (162) <223> Xaa equals any of the naturally occurring L-amino acids <400> 125 Gly Val Asn His Glu Ser Trp Arg Asp Ser Gly Gln Thr Asp Gly Leu Thr Phe Gly His Leu Lys Met Val Leu Leu Trp Ala Ser Val Leu Phe Pro Ala Pro Glu Asp Trp Ala Glu Leu Gln Gly Ala Val Tyr Arg Leu Leu Val Val Leu Leu Cys Cys Leu Ala Thr Arg Lys Leu Pro His Phe Leu His Pro Gln Arg Asn Leu Leu Gln Gly Ser Gly Leu Asp Leu Gly Ala Ile Tyr Gln Arg Val Glu Gly Phe Ala Ser Gln Pro Glu Ala Ala Leu Arg Ile His Ala Thr His Leu Gly Arg Ser Pro Pro Pro Arg Ile Gly Ser Gly Leu Lys Ala Leu Leu Gln Leu Pro Ala Ser Asp Pro Thr Tyr Trp Ala Thr Ala Tyr Phe Asp Val Leu Leu Asp Lys Phe Gln Val Phe Asn Ile Gln Asp Lys Asp Arg Ile Ser Ala Met Gln Ser Ile Phe Gln Xaa Thr Arg Thr Leu Gly Gly Glu Glu Ser <210> 126 <211> 205 <212> PRT
<213> Homo sapiens <400> 126 Met Gln Glu Leu Leu Gly Arg Gly Ala Trp Ala Pro Gly Cys Gly Arg Arg Pro Ser Gly~Trp Gly Gln Leu Ala Cys Pro Asp Pro Leu Leu Pro Pro His Asn Pro Lys Ser Pro Gln Pro Gly Pro Ser Thr Ser Gly Val Trp Gly Glu Glu Gln Gly Leu Arg Thr Leu Ser Ser Glu His Pro Trp Gln Gly Leu Gln Pro Leu Ile Ser Ser Leu Lys Pro Cys Gly His Thr Ala Arg Arg Asp Leu Pro Leu Ala Pro Ala Ser Phe Gln Pro Arg Val Leu Ile Gln Gly Pro Arg Thr Val Pro Pro Val Leu Leu Cys Pro Gln His Lys Ala Arg Leu His Ser Gln Lys Cys Ser Gln Ala Leu Glu Gly Asp Pro Ala Ser Ser Pro Thr Ala Pro His Pro Thr His Pro Ser Ala Ala Pro Leu Leu Phe Pro Arg Asp Leu Ser Tyr Thr Gly Gln Glu Ala Ala Glu Arg Val Ser Pro Pro Pro Ser Lys Arg Ser Cys Ser Leu Cys Gln Asn Arg Val Trp Ala Gly Gly Arg Ala Leu Gly Ala Arg Pro Leu Pro Leu Pro Ala Gly Phe Ser Trp Ser Leu Cys Trp Lys <210> 127 <211> 142 <212> PRT
<213> Homo sapiens <400> 127 Met Phe His Thr Phe Thr Leu Gln Gly Arg Gly Glu Gly Ser Cys Ser Pro Ser His Thr Ala Leu Leu Ser Asn Ile Thr Ala Pro Val Thr Thr Thr Leu Ser Arg Ala Leu Glu Gly Glu Gly Ser Met Pro Glu Pro Leu Ala Ala Gly Gly Pro Trp Arg Ser Ala Gly Pro Trp Trp Thr Arg Gln Glu Ile Cys Thr Pro Trp Ala Val Pro Ala His Thr Trp Gly Ser Arg Glu Asp Gly Ala Arg Ala Ser Leu His Ile Tyr Ala Ser Ser Arg Thr Gln Gly Ala Gly Pro Gly Ala Gly Pro Gly Arg Pro Gly Gly Ala Gly Gly Pro Gly Ala Ala Pro Gln Pro Pro Cys Leu Ala Gly Pro Glu Ser Ser His Thr Gly Ala Val Cys Leu Cys Trp Ser Gly Ala Ser <210> 128 <211> 152 <212> PRT
<213> Homo sapiens <400> 128 Met Gly Arg Glu Ala Thr Gly Gln Lys Ala Pro Gly Gly His Asp Leu Leu Ala Lys Arg Phe Gln Gln Ser Ala Lys Val Pro Ser Ser Ala Pro Gly Leu Pro Cys Leu Gly Pro Ser Pro Lys Asp Ala Ser Leu Cys Gly Trp Gly Val Pro Ala Gly Leu Gly Gly Ser Ser Val Leu Ser Cys Lys 50 . 55 60 Ser Val Cys Val Arg Val Ala Gly Thr Gly Gly Pro Thr Asp Gln Arg Ala Ala Val Gly Arg Ala Phe Gln Cys Ile Ala Arg Gly Lys Glu Leu Thr Leu Thr Val Pro Ser Val Ser Leu Val Gly Val Gly Gly Phe Ser 1'00 105 110 Leu Arg Gly Cys Phe Thr His Ser His Cys Arg Gly Gly Ala Arg Gly Ala Ala Leu Pro Leu Thr Gln Leu Tyr Cys Gln Thr Ser Arg His Gln Ser Pro Gln Pro Cys His Val His <210> 129 <211> 471 <212> PRT
<213> Homo sapiens <400> 129 Pro Arg Val Arg Ser Glu Ala Ala Leu Ser Ser Gln Arg Asp Leu Phe Ser Ser Phe Phe Leu Asn Ser Ser Ser Arg Glu Arg Leu Val Val Thr Gly Arg Ala Gly Trp Met Gly Met Gly Arg Gly Ala Gly Arg Ser Ala Leu Gly Phe Trp Pro Thr Leu Ala Phe Leu Leu Cys Ser Phe Pro Ala Ala Thr Ser Pro Cys Lys Ile Leu Lys Cys Asn Ser Glu Phe Trp Ser Ala Thr Ser Gly Ser His Ala Pro Ala Ser Asp Asp Thr Pro Glu Phe Cys Ala Ala Leu Arg Ser Tyr Ala Leu Cys Thr Arg Arg Thr Ala Arg Thr Cys Arg Gly Asp Leu Ala Tyr His Ser Ala Val His Gly Ile Glu Asp Leu Met Ser Gln His Asn Cys Ser Lys Asp Gly Pro Thr Ser Gln Pro Arg Leu Arg Thr Leu Pro Pro Ala Gly Asp Ser Gln Glu Arg Ser Asp Ser Pro Glu Ile Cys His Tyr Glu Lys Ser Phe His Lys His Ser Ala Thr Pro Asn Tyr Thr His Cys Gly Leu Phe Gly Asp Pro His Leu Arg Thr Phe Thr Asp Arg Phe Gln Thr Cys Lys Val Gln Gly Ala Trp Pro Leu Ile Asp Asn Asn Tyr Leu Asn Val Gln Val Thr Asn Thr Pro Val Leu Pro Gly Ser Ala Ala Thr Ala Thr Ser Lys Leu Thr Ile Ile Phe Lys Asn Phe Gln Glu Cys Val Asp Gln Lys Val Tyr Gln Ala Glu Met Asp Glu Leu Pro Ala Ala Phe Val Asp Gly Ser Lys Asn Gly Gly Asp Lys His Gly Ala Asn Ser Leu Lys Ile Thr Glu Lys Val Ser Gly Gln His Val Glu Ile Gln Ala Lys Tyr Ile Gly Thr Thr Ile Val Val 290 295 300.
Arg Gln Val Gly Arg Tyr Leu Thr Phe Ala Val Arg Met Pro Glu Glu Val Val Asn Ala Val Glu Asp Trp Asp Ser Gln Gly Leu Tyr Leu Cys Leu Arg Gly Cys Pro Leu Asn Gln Gln Ile Asp Phe Gln Ala Phe His Thr Asn Ala Glu Gly Thr Gly Ala Arg Arg Leu Ala Ala Ala Ser Pro Ala Pro Thr Ala Pro Glu Thr Phe Pro Tyr Glu Thr Ala Val Ala Lys Cys Lys Glu Lys Leu Pro Val Glu Asp Leu Tyr Tyr Gln Ala Cys Val Phe Asp Leu Leu Thr Thr Gly Asp Val Asn Phe Thr Leu Ala Ala Tyr Tyr Ala Leu Glu Asp Val Lys Met Leu His Ser Asn Lys Asp Lys Leu His Leu Tyr Glu Arg Thr Arg Asp Leu Pro Gly Arg Ala Ala Ala Gly Leu Pro Leu Ala Pro Arg Pro Leu Leu Gly Ala Leu Val Pro Leu Leu Ala Leu Leu Pro Val Phe Cys <210> 130 <211> 164 <212> PRT
<213> Homo sapiens <400> 130 Met Arg His His Thr Trp Leu Ile Leu Tyr Ile Tyr Phe Leu Tyr Phe Gln Glu Gly Val Leu Pro Cys Cys Pro Gly Trp Ser Ala Val Ala Arg 20 . 25 30 Ser Trp Leu Thr Ala Thr Ser Ala Phe Arg Val Gln Ala Val Leu Cys Leu Ser Leu Pro Ser Ser Trp Asp Tyr Arg Cys Pro Pro Pro His Pro Ala Asn Phe Cys Val Cys Val Cys Val Phe Leu Val Glu Met Gly Phe His His Leu Gly Gln Ala Gly Leu Glu Leu Leu Thr Leu Asp Pro Pro Val Ser Ala Ser Gln Ser Asp Gly Ile Thr Gly Val Ser His Cys Ala Gln Pro Asn Phe Val Tyr Leu Val Glu Met Gly Phe Leu His Val Gly Gln Ala Gly Leu Lys Leu Pro Asn Leu Gly Asp Pro Pro Ala Ser Ala Ser Gln Ser Ala Gly Ile Thr Asp His Glu Val Arg Ser Ser Arg Leu Ala Trp Pro Thr <210> 131 <211> 311 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (125) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (126) <223> Xaa equals any of the naturally occurring L-amino acids <400> 131 Arg Thr Arg Gly Gly Arg Val Trp Thr Ser Val Leu Leu Phe Ala Phe Ala Thr Gly Val Arg Ser Asn Gly Leu Val Ser Val Gly Phe Leu Met 20 ~ 25 30 His Ser Gln Cys Gln Gly Phe Phe Ser Ser Leu Thr Met Leu Asn Pro Leu Arg Gln Leu Phe Lys Leu Met Ala Ser Leu Phe Leu Ser Val Phe Thr Leu Gly Leu Pro Phe Ala Leu Phe Gln Tyr Tyr Ala Tyr Thr Gln Phe Cys Leu Pro Gly Ser Ala Arg Pro Ile Pro Glu Pro Leu Val Gln Leu Ala Val Asp Lys Gly Tyr Arg Ile Ala Glu Gly Asn Glu Pro Pro Trp Cys Phe Trp Asp Val Pro Leu Ile Tyr Ser Tyr Xaa Xaa Asp Val Tyr Trp Asn Val Gly Phe Leu Lys Tyr Tyr Glu Leu Lys Gln Val Pro Asn Phe Leu Leu Ala Ala Pro Val Ala Ile Leu Val Ala Trp Ala Thr Trp Thr Tyr Val Thr Thr His Pro Trp Leu Cys Leu Thr Leu Gly Leu Gln Arg Ser Lys Asn Asn Lys Thr Leu Glu Lys Pro Asp Leu Gly Phe Leu Ser Pro Gln Val Phe Val Tyr Val Val His Ala Ala Val Leu Leu Leu Phe Gly Gly Leu Cys Met His Val Gln Val Leu Thr Arg Phe Leu Gly Ser Ser Thr Pro Ile Met Tyr Trp Phe Pro Ala His Leu Leu Gln Asp Gln Glu Pro Leu Leu Arg Ser Leu Lys Thr Val Pro Trp Lys Pro Leu Ala Glu Asp Ser Pro Pro Gly Gln Lys Val Pro Arg Asn Pro Ile 260 265 . 270 Met Gly Leu Leu Tyr His Trp Lys Thr Cys Ser Pro Val Thr Arg Tyr Ile Leu Gly Tyr Phe Leu Thr Tyr Trp Leu Leu Gly Leu Leu Leu His Cys Asn Phe Leu Pro Trp Thr <210> 132 <211> 111 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (75) <223> Xaa equals any of the naturally occurring L-amino acids <400> 132 Arg His Thr His Gln Gly Met Phe Ala Leu Ala Trp Lys Val Ile Phe Ser Val Met Leu Gln Asn Pro Ile Arg Tyr Pro Ser Val Leu Gly Ile Lys Ser Ser Leu Leu Ser Ser Leu Val Leu Val Met Val Trp Gly Asn Glu Lys Ser Gly Pro Cys Pro Thr Pro Lys Ser Arg Lys Gly Arg Arg Ser Cys Pro Ala Gln Val Gly Arg Gly Glu Xaa Gly Ser Tyr Trp Asp Pro Glu Phe Arg Leu Ser Arg Lys Ser Asn Gln Gly Leu Arg Arg Asp Tyr Leu Ser Leu Tyr His Phe Asn Leu His Phe Arg Asp Thr Phe <210> 133 <211> 119 <212> PRT
<213> Homo sapiens <400> 133 Met Gln Glu Phe His Leu Leu His Pro Val Pro Leu Ile Gln Gln Glu Thr Phe Ala Pro Gln Val Leu Ala Val Gln Gly Ser Pro Asn Ser Ala Ser Val Arg Leu Glu Cys Val Ser Lys Met Gln Val Glu Val Ile Gln Ala Gln Val Ile Pro Pro Gln Pro Leu Val Gly Phe Pro Ala Gln Ser Glu Phe Trp Val Pro Ile Gly Pro Leu Phe Ser Pro Pro His Leu Cys Trp Thr Ala Pro Ser Ser Leu Pro Gly Leu Trp Cys Gly Ala Gly Pro Thr Leu Phe Ile Ser Pro Asn His His Gln His Lys Thr Thr Glu Gln Gly Gly Leu Tyr Thr Lys Asp <210> 134 <211> 103 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (10) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (21) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (23) <223> Xaa equals any of the naturally occurring L-amino acids <400> 134 Met Thr Ser His Ala Arg Val Arg Lys Xaa Gly Ser Ser Arg Ala Ala Ala Leu Phe Phe Xaa Phe Xaa Phe Phe Phe Phe Phe Leu Lys Arg Pro Glu Ser Cys Ser Val Pro Gln Pro Gly Val Gln Trp Cys Asn Leu Gly Ser Leu Gln Leu Pro Pro Pro Arg Phe Lys Arg Phe Ser Cys Leu Ser Leu Pro Gly Ser Trp Asp Tyr Arg Cys Thr Pro Pro Arg Leu Ala Asn Phe Leu Ile Phe Ser Arg Asp Arg Val Ser Pro Cys Trp Pro Gly Trp Ser Arg Thr Pro Asp Leu Lys <210> 135 <211> 127 <212> PRT
<213> Homo Sapiens <400> 135 Glu Tyr Ala Ser Ser Asn Thr Thr His Tyr Arg Glu Ser Trp Tyr Ala Cys Arg Tyr Arg Ser Gly Ile Pro Gly Leu Leu Ile Phe Pro Gly Pro Gly Thr Gly Met Thr Ser Leu Leu Glu Gly Arg Met Val Leu Cys Val Ser Cys Leu Leu Leu Pro Leu Leu Leu Leu Leu Lys His Phe Asn Gly Leu Met Thr Pro Tyr Leu Ala His Asn Val Tyr Cys Pro Ile Glu Tyr Ile Ser Phe Phe Pro Phe His Glu Lys Asn Ile Glu Tyr Ile Ser Ile Trp Phe Ile Phe Asp Ser Phe Lys Phe Ile Tyr Ser Arg Leu Leu Cys Ile Ser Gln Ile Tyr Val Leu Tyr Arg Ala Tyr Thr Leu Pro His <210> 136 <211> 464 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (166) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (307) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (312) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (351) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (462) <223> Xaa equals any of the naturally occurring L-amino acids <400> 136 Asn Ser Ala Arg Gly Gly Ala Gly Pro Ala Val Ala Arg Asp Ser Leu Gln Val Thr Met Leu Trp Phe Ser Gly Val Gly Ala Leu Ala Glu Arg Tyr Cys Arg Arg Ser Pro Gly Ile Thr Cys Cys Val Leu Leu Leu Leu Asn Cys Ser Gly Val Pro Met Ser Leu Ala Ser Ser Phe Leu Thr Gly Ser Val Ala Lys Cys Glu Asn Glu Gly Glu Val Leu Gln Ile Pro Phe Ile Thr Asp Asn Pro Cys Ile Met Cys Val Cys Leu Asn Lys Glu Val Thr Cys Lys Arg Glu Lys Cys Pro Val Leu Ser Arg Asp Cys Ala Leu Ala Ile Lys Gln Arg Gly Ala Cys Cys Glu Gln Cys Lys Gly Cys Thr Tyr Glu Gly Asn Thr Tyr Asn Ser Ser Phe Lys Trp Gln Ser Pro Ala Glu Pro Cys Val Leu Arg Gln Cys Gln Glu Gly Val Val Thr Glu Ser Gly Val Arg Cys Val Xaa His Cys Lys Asn Pro Leu Glu His Leu Gly Met Cys Cys Pro Thr Cys Pro Gly Cys Val Phe Glu Gly Val Gln Tyr Gln Glu Gly Glu Glu Phe Gln Pro Glu Gly Ser Lys Cys Thr Lys Cys Ser Cys Thr Gly Gly Arg Thr Gln Cys Val Arg Glu Val Cys Pro Ile Leu Ser Cys Pro Gln His Leu Ser His Ile Pro Pro Gly Gln Cys Cys Pro Lys Cys Leu Gly Gln Arg Lys Val Phe Asp Leu Pro Phe Gly Ser Cys Leu Phe Arg Ser Asp Val Tyr Asp Asn Gly Ser Ser Phe Leu Tyr Asp Asn Cys Thr Ala Cys Thr Cys Arg Asp Ser Thr Val Val Cys Lys Arg Lys Cys Ser His Pro Gly Gly Cys Asp Gln Gly Gln Glu Gly Cys Cys Glu Xaa Cys Leu Leu Arg Xaa Pro Pro Glu Asp Ile Lys Val Cys Lys Phe Gly Asn Lys Ile Phe Gln Asp Gly Glu Met Trp Ser Ser Ile Asn Cys Thr Ile Cys Ala Cys Val Lys Gly Arg Thr Glu Cys Xaa Asn Lys Gln Cys Ile Pro Ile Ser Ser Cys Pro Gln Gly Lys Ile Leu Asn Arg Lys Gly Cys Cys Pro Ile Cys Thr Glu Lys Pro Gly Val Cys Thr Val Phe Gly Asp Pro His Tyr Asn Thr Phe Asp Gly Arg Thr Phe Asn Phe Gln Gly Thr Cys Gln Tyr Val Leu Thr Lys Asp Cys Ser Ser Pro Ala Ser Pro Phe Gln Val Leu Val Lys Asn Asp Ala Arg Arg Thr Arg Ser Phe Ser Trp Thr Lys Ser Val Glu Leu Val Leu Gly Glu Thr Gly Ser Ala Cys Ser Ser Thr Ser Pro Cys Ala Gly Thr Ala Xaa Ala Ser <210> 137 <211> 273 <212> PRT
<213> Homo sapiens <400> 137 Met Leu Phe Phe Cys Gly Asp Leu Leu Ser Arg Ser Gln Ile Phe Tyr Tyr Ser Thr Gly Met Thr Val Gly Ile Val Ala Ser Leu Leu Ile Ile Ile Phe Ile Leu Ser Lys Phe Met Pro Lys Lys Ser Pro Ile Tyr Val Ile Leu Val Gly Gly Trp Ser Phe Ser Leu Tyr Leu Ile Gln Leu Val Phe Lys Asn Leu Gln Glu Ile Trp Arg Cys Tyr Trp Gln Tyr Leu Leu Ser Tyr Val Leu Thr Val Gly Phe Met Ser Phe Ala Val Cys Tyr Lys Tyr Gly Pro Leu Glu Asn Glu Arg Ser Ile Asn Leu Leu Thr Trp Thr Leu Gln Leu Met Gly Leu Cys Phe Met Tyr Ser Gly Ile Gln Ile Pro His Ile Ala Leu Ala Ile Ile Ile Ile Ala Leu Cys Thr Lys Asn Leu Glu His Pro Ile Gln Trp Leu Tyr Ile Thr Cys Arg Lys Val Cys Lys Gly Ala Glu Lys Pro Val Pro Pro Arg Leu Leu Thr Glu Glu Glu Tyr Arg Ile Gln Gly Glu Val Glu Thr Arg Lys Ala Leu Glu Glu Leu Arg Glu Phe Cys Asn Ser Pro Asp Cys Ser Ala Trp Lys Thr Val Ser Arg Ile Gln Ser Pro Lys Arg Phe Ala Asp Phe Val Glu Gly Ser Ser His Leu Thr Pro Asn Glu Val Ser Val His Glu Gln Glu Tyr Gly Leu Gly Ser Ile Ile Ala Gln Asp Glu Ile Tyr Glu Glu Ala Ser Ser Glu Glu Glu Asp Ser Tyr Ser Arg Cys Pro Ala Ile Thr Gln Asn Asn Phe Leu Thr <210> 138 <211> 286 <212> PRT
<213> Homo sapiens <400> 138 Phe Asp Pro Lys Leu Phe Leu Val Phe Leu Leu Gly Leu Met Leu Phe Phe Cys Gly Asp Leu Leu Ser Arg Ser Gln Ile Phe Tyr Tyr Ser Thr Gly Met Thr Val Gly Ile Val Ala Ser Leu Leu Ile Ile Ile Phe Ile Leu Ser Lys Phe Met Pro Lys Lys Ser Pro Ile Tyr Val Ile Leu Val Gly Gly Trp Ser Phe Ser Leu Tyr Leu Ile Gln Leu Val Phe Lys Asn Leu Gln Glu Ile Trp Arg Cys Tyr Trp Gln Tyr Leu Leu Ser Tyr Val Leu Thr Val Gly Phe Met Ser Phe Ala Val Cys Tyr Lys Tyr Gly Pro Leu Glu Asn Glu Arg Ser Ile Asn Leu Leu Thr Trp Thr Leu Gln Leu 115 ~ 120 125 Met Gly Leu Cys Phe Met Tyr Ser Gly Ile Gln Ile Pro His Ile Ala 130 135 ~ 140 Leu Ala Ile Ile Ile Ile Ala Leu Cys Thr Lys Asri Leu Glu His Pro Ile Gln Trp Leu Tyr.Ile Thr Cys Arg Lys Val Cys Lys Gly Ala Glu Lys Pro Val Pro Pro Arg Leu Leu Thr Glu Glu Glu Tyr Arg Ile Gln Gly Glu Val Glu Thr Arg Lys Ala Leu Glu Glu Leu Arg Glu Phe Cys Asn Ser Pro Asp Cys Ser Ala Trp Lys Thr Val Ser Arg Ile Gln Ser Pro Lys Arg Phe Ala Asp Phe Val Glu Gly Ser Ser His Leu Thr Pro Asn Glu Val Ser Val His Glu Gln Glu Tyr Gly Leu Gly Ser Ile Ile Ala Gln Asp Glu Ile Tyr Glu Glu Ala Ser Ser Glu Glu Glu Asp Ser Tyr Ser Arg Cys Pro Ala Ile Thr Gln Asn Asn Phe Leu Thr <210> 139 <211> 560 <212> PRT
<213> Homo Sapiens <220>
<221> SITE
<222> (11) <223> Xaa equals any of the naturally occurring L-amino acids <220>
<221> SITE
<222> (20) <223> Xaa equals any of the naturally occurring L-amino acids <400> 139 Leu Arg Thr Arg Gly Ser Leu Thr Ser Trp Xaa Leu Cys Cys Val Trp Trp Arg Pro Xaa Leu Pro Ile Ser Ala Gly His Glu Glu Leu Ser Arg Leu Ser Gln Tyr Arg Lys Glu Leu Gly Ala Gln Leu His Gln Leu Val Leu Asn Ser Ala Arg Glu Lys Arg Asp Met Glu Gln Arg His Val Leu Leu Lys Gln Lys Glu Leu Gly Gly Glu Glu Pro Glu Pro Ser Leu Arg Glu Gly Pro Gly Gly Leu Val Met Glu Gly His Leu Phe Lys Arg Ala Ser Asn Ala Phe Lys Thr Trp Ser Arg Arg Trp Phe Thr Ile Gln Ser Asn Gln Leu Val Tyr Gln Lys Lys Tyr Lys Asp Pro Val Thr Val Val Val Asp Asp Leu Arg Leu Cys Thr Val Lys Leu Cys Pro Asp Ser Glu Arg Arg Phe Cys Phe Glu Val Val Ser Thr Ser Lys Ser Cys Leu Leu Gln Ala Asp Ser Glu Arg Leu Leu Gln Leu Trp Val Ser Ala Val Gln Ser Ser Ile Ala Ser Ala Phe Ser Gln Ala Arg Leu Asp Asp Ser Pro Arg Gly Pro Gly Gln Gly Ser Gly His Leu Ala Ile Gly Ser Ala Ala Thr Leu Gly Ser Gly Gly Met Ala Arg Gly Arg Glu Pro Gly Gly Val Gly His Val Val Ala Gln Val Gln Ser Val Asp Gly Asn Ala Gln Cys Cys Asp Cys Arg Glu Pro Ala Pro Glu Trp Ala Ser Ile Asn Leu Gly Val Thr Leu Cys Ile Gln Cys Ser Gly Ile His Arg Ser Leu Gly Val His Phe Ser Lys Val Arg Ser Leu Thr Leu Asp Ser Trp Glu Pro Glu Leu Val Lys Leu Met Cys Glu Leu Gly Asn Val Ile Ile Asn Gln Ile Tyr Glu Ala Arg Val Glu Ala Met Ala Val Lys Lys Pro Gly Pro Ser Cys Ser Arg Gln Glu Lys Glu Ala Trp Ile His Ala Lys Tyr Val Glu Lys Lys Phe Leu Thr Lys Leu Pro Glu Ile Arg Gly Arg Arg Gly Gly Arg Gly Arg Pro Arg Gly Gln Pro Pro Val Pro Pro Lys Pro Ser Ile Arg Pro Arg Pro Gly Ser Leu Arg Ser Lys Pro Glu Pro Pro Ser Glu Asp Leu Gly Ser Leu His Pro Gly Ala Leu Leu Phe Arg Ala Ser Gly His Pro Pro Ser Leu Pro Thr Met Ala Asp Ala Leu Ala His Gly Ala Asp Val Asn Trp Val Asn Gly Gly Gln Asp Asn Ala Thr Pro Leu Ile Gln Ala Thr Ala Ala Asn Ser Leu Leu Ala Cys Glu Phe Leu Leu Gln Asn Gly Ala Asn Val Asn Gln Ala Asp Ser Ala Gly Arg Gly Pro Leu His His Ala Thr Ile Leu Gly His Thr Gly Leu Ala Cys Leu Phe Leu Lys Arg Gly Ala Asp Leu Gly Ala Arg Asp Ser Glu Gly Arg Asp Pro Leu Thr Ile Ala Met Glu Thr Ala Asn Ala Asp Ile Val Thr Leu Leu Arg Leu Ala Lys Met Arg Glu Ala Glu Ala Ala Gln Gly Gln Ala Gly Asp Glu Thr Tyr Leu Asp Ile Phe Arg Asp Phe Ser Leu Met Ala Ser Asp Asp Pro Glu Lys Leu Ser Arg Arg Ser His Asp Leu His Thr Leu <210> 140 <211> 90 <212> PRT
<213> Homo sapiens <220>
<221> SITE
<222> (85) <223> Xaa equals any of the naturally occurring L-amino acids <400> 140 Gly Leu Arg Gly Phe Arg Glu Ala Arg Arg Asp Phe~Trp Arg Gly Ala Glu Ser Leu Glu Ala Ala Leu Thr His Asn Ala Glu Val Pro Arg Arg Arg Ala Gln Glu Ala Glu Glu Ala Gly Ala Ala Leu Arg Thr Ala Arg Ala Gly Tyr Arg Gly Arg Ala Leu Asp Tyr Ala Leu Gln Ile Asn Val Ile Glu Asp Lys Arg Lys Phe Asp Ile Met Glu Phe Val Leu Arg Leu Val Glu Ala Gln Xaa Thr His Phe Ser Arg <210> 141 <211> 234 <212> PRT
<213> Homo Sapiens <400> 141 Pro Thr Arg Pro Pro Thr Arg Pro Val Arg Val Ser Val Gly Gly Leu Val Gly Glu Val Ala Cys Ala Cys Arg Asp Cys Ile Pro Glu Thr Met Ala Glu Gly Asp Asn Arg Ser Thr Asn Leu Leu Ala Ala Glu Thr Ala Ser Leu Glu Glu Gln Leu Gln Gly Trp Gly Glu Val Met Leu Met Ala Asp Lys Val Leu Arg Trp Glu Arg Ala Trp Phe Pro Pro Ala Ile Met Gly Val Val Ser Leu Val Phe Leu Ile Ile Tyr Tyr Leu Asp Pro Ser Val Leu Ser Gly Val Ser Cys Phe Val Met Phe Leu Cys Leu Ala Asp Tyr Leu Val Pro Ile Leu Ala Pro Arg Ile Phe Gly Ser Asn Lys Trp Thr Thr Glu Gln Gln Gln Arg PY~,e His Glu Ile Cys Ser Asn Leu Val Lys Thr Arg Arg Arg Ala Val Gly Trp Trp Lys Arg Leu Phe Thr Leu Lys Glu Glu Lys Pro Lys Met Tyr Phe Met Thr Met Ile Val Ser Leu Ala Ala Val Ala Trp Val Gly Gln Gln Val His Asn Leu Leu Leu Thr Tyr Leu Ile Val Thr Ser Leu Leu Leu Leu Pro Gly Leu Asn Gln His Gly Ile Ile Leu Lys Tyr Ile Gly Met Ala Lys Arg Glu Ile Asn Lys Leu Leu Lys Gln Lys Glu Lys Lys Asn Glu <210> 142 <211> 75 <212> PRT
<213> Homo Sapiens <400> 142 Met Gly Ser Cys Tyr Ile Tyr Gln Pro Gln Tyr Leu Gly Ala Ala Leu Ser Arg Thr Asn Gly Leu Pro Ile Gly Asp Ala Ile Thr Gly Ser Val Leu Gly Gly Gly Val Leu Arg Gly Asp Gly His Leu Ala Gly Ser Ala Ser Pro Gly Asn Ser Ser His Pro Gln Ser Leu Thr Gln Gly Pro Arg Ser Cys Arg Leu Trp Ala Arg Ala Ala Arg Ala <210> 143 <211> 95 <212> PRT
<213> Homo Sapiens <400> 143 Met Pro Ala Leu Ala Leu Ser Pro Arg Pro Leu Leu Pro Gly Ile Ser Cys Pro Ser Pro Arg Cys Thr Cys Pro Ser Pro Ala Asn Pro Leu Cys Leu His Phe Thr Arg Leu Gln Gly Val His Ile Ser Ser Ser Asp Gln Gly Ser Glu Gly Arg Gly Cys Val Ser Cys Ile Ser Leu Gly Pro Leu Arg Gly Arg Asp Cys Phe Pro Arg Val Gly Ala Leu Glu Val Met Arg Leu Phe Pro Pro Asn His Gln Ile Trp Thr Glu Asn Cys Ile Gly <210> 144 <211> 234 <212> PRT
<213> Homo Sapiens <400> 144 Met Ala Thr Thr Thr Cys Val Pro Met Lys Thr Ala Arg Leu Pro Gly Gln Cys Ser Met Leu Thr Thr Val Pro Leu Ala Leu Pro Ala Ser Thr Ala Ala Pro His Pro Val Ser Pro Thr Pro Ala Ala Met Pro Ser Ser Thr Pro Ile Ala Gly Ala Leu Pro Leu Thr Met Ala Leu Ser Pro His Pro Leu Ala Leu Leu Pro Gln Leu Ala Leu Thr Gln Pro Ala Ser Cys Pro Thr Arg Thr Met Arg Ser Ser Thr Pro Ile Pro Ser Leu Gly Gln Leu Gly Thr Pro Pro Thr Asp Trp Ala Thr Pro Arg Pro His Pro Leu Ala Trp Ser Gly Pro His Met Arg Arg Met Thr Pro Leu Ala Ser Thr Pro Gln Pro Leu Asp Ser Pro Thr Pro Pro Ser Thr Arg Thr Thr Ala Ser Asp Ser Ser Ser Ala Cys Arg Leu Thr Cys Arg Gly Gln Ser Leu 145 ' 150 155 160 Ala Gly Ala Ser Leu Arg Gly Arg Gly Glu Gly Phe His Ser Cys Ser Leu Ile Phe Arg Gly Ile Ala His Cys Ser Leu Leu Gly Pro Ala Phe Phe Leu Pro Pro Trp Gln Val Gly Ser Arg Ser Pro Arg Asn Thr Pro Ser Arg Gly Trp Cys Ser Val His Ala Pro Ala Ser Trp Ala Cys Pro Ser Leu Phe Phe Gly Arg Met Cys Leu Phe <210> 145 <211> 131 <212> PRT
<213> Homo Sapiens <400> 145 Met Asp Arg Val Thr Val Gly Gly Trp Cys Thr Ser Ser Ser Met Glu Val Ser Leu Pro Leu Gly Met Ala Ser Phe Met Leu Arg Gln Val Lys Thr Arg Pro Met Ser Arg Ser Val Gly Leu Ile Ser Ser Trp Leu Thr Val Val Ser Leu Phe Pro Ser Phe Ser Asn Ser Val Leu Ala Thr Ala Pro Ser Cys Cys Val Pro Ser Arg Asn Gln Met Met Val Ala Ala Gly Phe Ala Leu Lys Ala Arg His Val Arg Leu Trp Gly Val Pro Ala Cys Ser Arg Ile Thr Gly Pro Pro Ser Ile Leu Val Ser Ser Gly Gly Met Ser Thr Val Ser Leu Ala Arg Arg Glu Arg Arg Ala Ala Ser Val Ala Trp His Ser <210>

<211>

<212>
PRT

<213>
Homo Sapiens <400>

His Glu Arg AlaLeuThr ThrMetGln IleGln ValAlaGly Leu Gln Leu Gln Ala ValProLeu PheSerThr AlaGlu GluAspLeu Leu Phe Ala Ile Leu LeuLeuAsn SerSerGlu SerSer LeuHisGln Leu Gln Thr Ala Val AspCysArg GlyLeuHis LysAsp TyrLeuAsp Ala Met Leu Ala Ile CysTyrAsp GlyLeuGln GlyLeu LeuTyrLeu Gly Gly Leu Phe Phe LeuAlaAla LeuAlaPhe SerThr MetIleCys Ala Ser Gly Pro Ala TrpLysHis PheThrThr ArgAsn ArgAspTyr Asp Arg Asp Ile Asp AspAspPro PheAsnPro GlnAla TrpArgMet Ala Asp Ala His Ser Pro Pro Arg Gly Gln Leu His Ser Phe Cys Ser Tyr Ser Ser Gly Leu Gly Ser Gln Thr Ser Leu Gln Pro Pro Ala Gln Thr Ile Ser Asn Ala Pro Val Ser Glu Tyr Met Asn Gln Ala Met Leu Phe Gly Arg Asn Pro Arg Tyr Glu Asn Val Pro Leu Ile Gly Arg Ala Ser Pro Pro Pro Thr Tyr Ser Pro Ser Met Arg Ala Thr Tyr Leu Ser Val Ala Asp Glu His Leu Arg His Tyr Gly Asn Gln Phe Pro Ala DEMANDE OU BREVET VOLUMINEUX
LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.

~~ TTENANT LES PAGES 284 A 436 NOTE : Pour les tomes additionels, veuillez contacter 1e Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS
THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE
VOLUME

NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME
NOTE POUR LE TOME / VOLUME NOTE:

Claims (23)

What Is claimed is:

1. An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of:

(a) a polynucleotide fragment of SEQ ID NO:X or a polynucleotide fragment of the cDNA sequence included in ATCC Deposit No:Z, which is hybridizable to SEQ ID NO:X;

(b) a polynucleotide encoding a polypeptide fragment of SEQ ID
NO:Y or a polypeptide fragment encoded by the cDNA sequence included in ATCC
Deposit No:Z, which is hybridizable to SEQ 117 NO:X;

(c) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y
or a polypeptide domain encoded by the cDNA sequence included in ATCC Deposit No:Z, which is hybridizable to SEQ ID NO:X;

(d) a polynucleotide encoding a polypeptide epitope of SEQ ID NO:Y
or a polypeptide epitope encoded by the cDNA sequence included in ATCC Deposit No:Z, which is hybridizable to SEQ ID NO:X;

(e) a polynucleotide encoding a polypeptide of SEQ ID NO:Y or the cDNA sequence included in ATCC Deposit No:Z, which is hybridizable to SEQ ID
NO:X, having biological activity;

(f) a polynucleotide which is a variant of SEQ ID NO:X;

(g) a polynucleotide which is an allelic variant of SEQ ID NO:X;

(h) a polynucleotide which encodes a species homologue of the SEQ
ID NO:Y;

(i) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(h), wherein said polynucleotide does not hybridize under stringent conditions to a nucleic acid molecule having a nucleotide sequence of only A residues or of only T residues.

2. The isolated nucleic acid molecule of claim 1, wherein the polynucleotide fragment comprises a nucleotide sequence encoding a secreted protein.

3. The isolated nucleic acid molecule of claim 1, wherein the polynucleotide fragment comprises a nucleotide sequence encoding the sequence identified as SEQ m NO:Y or the polypeptide encoded by the cDNA sequence included in ATCC Deposit No:Z, which is hybridizable to SEQ m NO:X.

4. The isolated nucleic acid molecule of claim 1, wherein the polynucleotide fragment comprises the entire nucleotide sequence of SEQ ID
NO:X
or the cDNA sequence included in ATCC Deposit No:Z, which is hybridizable to SEQ ID NO:X.

5. The isolated nucleic acid molecule of claim 2, wherein the nucleotide sequence comprises sequential nucleotide deletions from either the C-terminus or the N-terminus.

6. The isolated nucleic acid molecule of claim 3, wherein the nucleotide sequence comprises sequential nucleotide deletions from either the C-terminus or the N-terminus.

7. A recombinant vector comprising the isolated nucleic acid molecule of claim 1.

8. A method of making a recombinant host cell comprising the isolated nucleic acid molecule of claim 1.

9. A recombinant host cell produced by the method of claim 8.

10. The recombinant host cell of claim 9 comprising vector sequences.

11. An isolated polypeptide comprising.an amino acid sequence at least 95% identical to a sequence selected from the group consisting of:

(a) a polypeptide fragment of SEQ ID NO:Y or the encoded sequence included in ATCC Deposit No:Z;

(b) a polypeptide fragment of SEQ ID NO:Y or the encoded sequence included in ATCC Deposit No:Z, having biological activity;

(c) a polypeptide domain of SEQ ID NO:Y or the encoded sequence included in ATCC Deposit No:Z;

(d) a polypeptide epitope of SEQ ID NO:Y or the encoded sequence included in ATCC Deposit No:Z;

(e) a secreted form of SEQ 1D NO:Y or the encoded sequence included in ATCC Deposit No:Z;

(f) a full length protein of SEQ ID NO:Y or the encoded sequence included in ATCC Deposit No:Z;

(g) a variant of SEQ ID NO:Y;

(h) an allelic variant of SEQ ID NO:Y; or (i) a species homologue of the SEQ ID NO:Y.

12. The isolated polypeptide of claim 11, wherein the secreted form or the full length protein comprises sequential amino acid deletions from either the C-terminus or the N-terminus.

13. An isolated antibody that binds specifically to the isolated polypeptide of claim 11.

l4. A recombinant host cell that expresses the isolated polypeptide of claim 11.

15. A method of making an isolated polypeptide comprising:

(a) culturing the recombinant host cell of claim 14 under conditions such that said polypeptide is expressed; and (b) recovering said polypeptide.

16. The polypeptide produced by claim 15.

17. A method for preventing, treating, or ameliorating a medical condition, comprising administering to a mammalian subject a therapeutically effective amount of the polypeptide of claim 11 or the polynucleotide of claim 1.

18. A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising:

(a) determining the presence or absence of a mutation in the polynucleotide of claim 1; and (b) diagnosing a pathological condition or a susceptibility to a pathological condition based on the presence or absence of said mutation.

19. A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising:

(a) determining the presence or amount of expression of the polypeptide of claim 11 in a biological sample; and (b) diagnosing a pathological condition or a susceptibility to a pathological condition based on the presence or amount of expression of the polypeptide.

20. A method for identifying a binding partner to the polypeptide of claim 11 comprising:

(a) contacting the polypeptide of claim 11 with a binding partner; and (b) determining whether the binding partner effects an activity of the polypeptide.

21. The gene corresponding to the cDNA sequence of SEQ ID NO:Y.

22. A method of identifying an activity.in a biological assay, wherein the method comprises:

(a) expressing SEQ ID NO:X in a cell;

(b) isolating the supernatant;

(c) detecting an activity in a biological assay; and (d) identifying the protein in the supernatant having the activity.

23. The product produced by the method of claim 20.