CA2265554A1 - 3' genomic promoter region and polymerase gene mutations responsible for attenuation in viruses of the order designated mononegavirales - Google Patents

Abstract

Isolated, recombinantly-generated, attenuated, nonsegmented, negative-sense, single stranded RNA viruses of the Order Mononegavirales having at least one attenuating mutation in the 3' genomic promoter region and having at least one attenuating mutation in the RNA polymerase gene are described. Vaccines are formulated comprising such viruses and a physiologically acceptable carrier.
The vaccines are used for immunizing an individual to induce protection against a nonsegmented, negative-sense, single stranded RNA virus of the Order Mononegavirales.

Description

?CA 02265554 l999-03- 16DEMANDES OU BREVETS VOLUMINEUXLA. PRESENTE PARTIE DE cerre DEMANDE ou cs BREVEFcompnemo PLUS D’UN TOME. - OCECI EST LE TOME A 057/T.NOTE: Pour les tomes additionels, veuillez contacter le Bureau canadien desbrevetsJUMBO AP.Pl_-_lC_ATlONS_/PATENTSTHIS SECTION OF THE APPLICATIONIEATENT CONTAINS MOREI-THAN ONE VOLUME 'THIS IS VOLUME _. _/__ OF 2NOTE: ‘For additionai volumes-please contactlthé Canadian Patent Office?10152025........._.._.a....._....z..l- . .W0 98/ 13501CA 02265554 l999-03- 16PCTIU S97] 167 183' GENOMIC PROMOTER REGION AND POLYMERASE GENEMUTATIONS RESPONSIBLE FOR ATTENUATION IN VIRUSESOF THE ORDER DESIGNATED MONONEGAVIRALESField Of The InventionThis invention relates to isolated,recombinantly-generated, attenuated, nonsegmented,negative-sense, single stranded RNA viruses of theOrder designated Mononegavirales having at least oneattenuating mutation in the 3' genomic promoter regionand having at least one attenuating mutation in the RNApolymerase gene. This invention was made withGovernment support under a grant awarded by the PublicHealth Service. The Government has certain rights inthe invention.Background Of The InventionEnveloped, negative—sense, single strandedRNA viruses are uniquely organized and expressed. Thegenomic RNA of negative-sense, single stranded virusesserves two template functions in the context of anucleocapsidz as a template for the synthesis ofmessenger RNAs (mRNAs) and as a template for thesynthesis of the antigenome (+) strand. Negative-sense, single stranded RNA viruses encode and packagetheir own RNA dependent RNA Polymerase. Messenger RNAsare only synthesized once the virus has been uncoatedin the infected cell. Viral replication occurs aftersynthesis of the mRNAs and requires the continuoussynthesis of viral proteins. The newly synthesizedantigenome (+) strand serves as the template forgenerating further copies of the (—) strand genomicRNA.SUBSTHUTESHEET(RULE26)..-....._.,..............m........._......... . . . _......——-nu... , . ,, ,........................._. ...~—.....—.«...,»...-—-~»»-?101520253035WO 98/13501CA 02265554 l999-03- 16PCT/U S97/ 16718The polymerase complex actuates and achievestranscription and replication by engaging the cis-acting signals at the 3' end of the genome, inparticular, the promoter region. Viral genes are thentranscribed from the genome template unidirectionallyfrom its 3' to its 5' end. There is always less mRNAmade from the downstream genes (e.g., the polymerasegene (L)) relative to their upstream neighbors (i.e.,the nucleoprotein gene (N)). Therefore, there is alwaysa gradient of mRNA abundance according to the positionof the genes relative to the 3'-end of the genome.Based on the revised reclassification in 1993by the International Committee on the Taxonomy ofViruses, an Order, designated Mononegavirales, has beenestablished. This Order contains three families ofenveloped viruses with single stranded, nonsegmentedRNA genomes of minus polarity (negative-sense). Thesefamilies are the Paramyxoviridae, Rhabdoviridae andFiloviridae. The family Paramyxoviridae has beenfurther divided into two subfamilies, Paramyxovirinaeand Pneumovirinae. The subfamily Paramyxovirinaecontains three genera, Paramyxovirus, Rubulavirus and'Mbrbillivirus. The subfamily Pneumovirinae containsthe genus Pneumovirus.The new classification is based uponmorphological criteria, the organization of the viralgenome, biological activities and the sequencerelationships of the proteins. The morphologicaldistinguishing feature among enveloped viruses for thesubfamily Paramyxovirinae is the size and shape of thenucleocapsids (diameter 18mm, 1mm in length, pitch of5.5 nm), which have a left—handed helical symmetry. Thebiological criteria are: 1) antigenic cross-reactivitybetween members of a genus, and 2) the presence ofneuraminidase activity in the genera Paramyxovirus,SUBSTHUTESHEET(RULE26)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718Rubulavirus and its absence in genus Mb??illivirus. Inaddition, variations in the coding potential of the Pgene are considered, as is the presence of an extragene (SH) in Rubulaviruses.Pneumoviruses can be distinguished fromParamyxovirinae morphologically because they containnarrow nucleocapsids. In addition, pneumoviruses havemajor differences in the number of protein—encodingcistrons (10 in pneumoviruses versus 6 inParamyxovirinae) and an attachment protein (G) that isvery different from that of Paramyxovirinae. Althoughthe paramyxoviruses and pneumoviruses have six proteinsthat appear to correspond in function (N, P, M, G/H/HN,F and L), only the latter two proteins exhibitsignificant sequence relatedness between the twosubfamilies. Several pneumoviral proteins lackcounterparts in most of the paramyxoviruses, namely thenonstructural proteins NS1 and NS2, the smallhydrophobic protein SH, and a second protein M2. Someparamyxoviral proteins, namely C and V, lackcounterparts in pneumoviruses. However, the basicgenomic organization of pneumoviruses andparamyxoviruses is the same. The same is true ofrhabdoviruses and filoviruses. Table 1 presents thecurrent taxonomical classification of these viruses,together with examples of each genus.Table 1Classification of Nonsegmented, negative-sense, singlestranded RNA Viruses of the Order MononegaviralesFamily ParamyxoviridaeSubfamily ParamyxovirinaeGenus ParamyxovirusSendai virus (mouse parainfluenza virustype 1)SUBSTITUTE SHEET (RULE 26)?101520253035ICA 02265554 l999-03- 16W0 98/13501 PCT/US97/16718Human parainfluenza virus (PIV) types 1and 3Bovine parainfluenza virus (BPV) type 3Genus RubulavirusSimian virus 5 (SV) (Canineparainfluenza virus type 2)Mumps virusNewcastle disease virus (NDV) (avianParamyxovirus 1)Human parainfluenza virus types 2, 4aand 4byGenus MbrbillivirusMeasles virus (MV)Dolphin MorbillivirusCanine distemper virus (CDV)Peste—des-petits—ruminants virusPhocine distemper virusRinderpest virussubfamily PneumovirinaeGenus PneumovirusHuman respiratory syncytial virus (RSV)Bovine respiratory syncytial virusPneumonia virus of miceTurkey rhinotracheitis virusFamily RhabdoviridaeGenus LyssavirusRabies virusGenus VesiculovirusVesicular stomatitis virusGenus EphemerovirusBovine ephemeral fever virusFamily FilovirdaeGenus FilovirusMarburg virusSUBSTITUTE SHEET (RULE 25)9?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/ 16718For many of these viruses, no vaccines of anykind are available. Thus, there is a need to developvaccines against such human and animal pathogens. Suchvaccines would have to elicit a protective immuneresponse in the recipient. The qualitative andquantitative features of such a favorable response areextrapolated from those seen in survivors of naturalvirus infection, who, in general, are protected fromreinfection by the same or highly related viruses forsome significant duration thereafter.A variety of approaches can be considered inseeking to develop such vaccines, including the use of:(1) purified individual viral protein vaccines (subunitvaccines); (2) inactivated whole virus preparations;and (3) live, attenuated viruses.Subunit vaccines have the desirable featureof being pure, definable and relatively easily producedin abundance by various means, including recombinantDNA expression methods. To date, with the notableexception of hepatitis B surface antigen, viral subunitvaccines have generally only elicited short—livedand/or inadequate immunity, particularly in naiverecipients.Formalin inactivated whole virus preparationsof polio (IPV) and hepatitis A have proven safe andefficacious. In contrast, immunization with similarlyinactivated whole viruses such as respiratory syncytialvirus and measles virus vaccines elicited unfavorableimmune responses and/or response profiles whichpredisposed vaccinees to exaggerated or aberrantdisease when subsequently confronted with the naturalor "wild-type" virus.Early attempts (1966) to vaccinate youngchildren using a parenterally administered formalin-inactivated RSV vaccine. Unfortunately, several fieldSUBSTITUTE SHEET (RULE 26)?1015202530W0 98/13501CA 02265554 l999-03- 16PCT/US97l16718trials of this vaccine revealed serious adversereactions -- the development of a severe illness withunusual features following subsequent natural infectionwith RSV (Bibliography entries 1,2). It has beensuggested that this formalinized RSV antigen elicitedan abnormal or unbalanced immune response profile,predisposing the vaccines to RSV disease (3,4).Thereafter, live, attenuated RSV vaccinecandidates were generated by cold passage or chemicalmutagenesis. These RSV strains were found to havereduced virulence in seropositive adults.Unfortunately, they proved either over or under-attenuated when given to seronegative infants; in somecases, they also were found to lack genetic stability(5,6). Another vaccination approach using parenteraladministration of live virus was ineffective andefforts along this line were discontinued (7).Notably, these live RSV vaccines were never associatedwith disease enhancement as observed with the formalin-inactivated RSV vaccine described above. Currently,there are no RSV vaccines approved for administrationto humans, although clinical trials are now in progresswith cold—passaged, chemically mutagenized strains ofRSV designated A2 and B-1.Appropriately attenuated live derivatives ofwild—type viruses offer a distinct advantage as vaccinecandidates. As live, replicating agents, they initiateinfection in recipients during which viral geneproducts are expressed, processed and presented in thecontext of the vaccinee's specific MC class I and IImolecules, eliciting humoral and cell-mediated immuneresponses, as well as the coordinate cytokine patterns,which parallel the protective immune profile ofsurvivors of natural infection.SUBSTITUTE SHEET (RULE 26)t?1015202530WO 98/13501CA 02265554 l999-03- 16PCT/US97/16718This favorable immune response pattern iscontrasted with the delimited responses elicited byinactivated or subunit vaccines, which typically arelargely restricted to the humoral immune surveillancearm. Further, the immune response profile elicited bysome formalin inactivated whole virus vaccines, e.g.,measles and respiratory syncytial virus vaccinesdeveloped in the 1960's, have not only failed toprovide sustained protection, but in fact have led to apredisposition to aberrant, exaggerated, and even fatalillness, when the vaccine recipient later confrontedthe wild-type virus.While live, attenuated viruses have highlydesirable characteristics as vaccine candidates, theyhave proven to be difficult to develop. The crux ofthe difficulty lies in the need to isolate a derivativeof the wild—type virus which has lost its disease-producing potential (i.e., virulence), while retainingsufficient replication competence to infect therecipient and elicit the desired immune responseprofile in adequate abundance.Historically, this delicate balance betweenvirulence and attenuation has been achieved by serialpassage of a wild—type viral isolate through differenthost tissues or cells under varying growth conditions(such as temperature). This process presumably favorsthe growth of viral variants (mutants), some of whichhave the favorable characteristic of attenuation.Occasionally, further attenuation is achieved throughchemical mutagenesis as well.This propagation/passage scheme typicallyleads to the emergence of virus derivatives which aretemperature sensitive, cold-adapted and/or altered intheir host range —- one or all of which are changesSUBSTWUTESHEET(RULE26)?1015202530W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718from the wild-type, disease-causing viruses —— i.e.,changes that may be associated with attenuation.Several live virus vaccines, including thosefor the prevention of measles and mumps (which areparamyxoviruses), and for protection against polio andrubella (which are positive strand RNA viruses), havebeen generated by this approach and provide themainstay of current childhood immunization regimensthroughout the world.Nevertheless, this means for generatingattenuated live virus vaccine candidates is lengthyand, at best, unpredictable, relying largely on theselective outgrowth of those randomly occurring genomicmutants with desirable attenuation characteristics.The resulting viruses may have the desired phenotype invitro, and even appear to be attenuated in animalmodels. However, all too often they remain eitherunder- or overattenuated in the human or animal hostfor whom they are intended as vaccine candidates.Even as to current vaccines in use, there isstill a need for more efficacious vaccines. Forexample, the current measles vaccines providereasonably good protection. However, recent measlesepidemics suggest deficiencies in the efficacy ofcurrent vaccines. Despite maternal immunization, highrates of acute measles infection have occurred inchildren under age one, reflecting the vaccines‘inability to induce anti-measles antibody levelscomparable to those developed following wild-typemeasles infection (8,9,10). As a result, vaccine-immunized mothers are less able to provide theirinfants with sufficient transplacentally-derivedpassive antibodies to protect the newborns beyond thefirst few months of life.SUBSTITUTE SHEET (RULE 26)v?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/ 16718Acute measles infections in previouslyimmunized adolescents and young adults point to anadditional problem. These secondary vaccine failuresindicate limitations in the current vaccines‘ abilityto induce and maintain antiviral protection that isboth abundant and long-lived (1l,12,13). Recently, yetanother potential problem was revealed. Thehemagglutinin protein of wild-type measles isolatedover the past 15 years has shown a progressivelyincreasing distance from the vaccine strains (14).This "antigenic drift" raises legitimate concerns thatthe vaccine strains may not contain the ideal antigenicrepertoire needed to provide optimal protection. Thus,there is a need for improved vaccines.Rational vaccine design would be assisted bya better understanding of these viruses, in particular,by the identification of the virally encodeddeterminants of virulence as well as those genomicchanges which are responsible for attenuation.Summary Of The InventionAccordingly, it is an object of thisinvention to identify those regions of the genome ofthe RNA viruses of the Order Mononegavirales wheremutations result in attenuation of those viruses.It is a further object of this invention toproduce recombinantly-generated viruses whichincorporate such attenuating mutations in theirgenomes.It is still a further object of thisinvention to formulate vaccines containing suchattenuated viruses.These and other objects of the invention asdiscussed below are achieved by the generation andSUBST?UTESHEET(RULE2Q?1015202530WO 98/13501CA 02265554 l999-03- 16PCT/US97/16718isolation of recombinantly-generated, attenuated,nonsegmented, negative-sense, single stranded RNAviruses of the Order Mononegavirales having at leastone attenuating mutation in the 3' genomic promoterregion and having at least one attenuating mutation inthe RNA polymerase gene.In the case of measles virus, at least oneattenuating mutation in the 3' genomic promoter regionis selected from the group consisting of nucleotide 26(A -9 T), nucleotide 42 (A -9 T or A -9 C) andnucleotide 96 (G -9 A), where these nucleotides, aswell as others delineated in this application (unlessstated otherwise), are presented in positive strand,antigenomic, that is, message (coding) sense, and atleast one attenuating mutation in the RNA polymerasegene is selected from the group consisting ofnucleotide changes which produce changes in an aminoacid selected from the group consisting of residues 331(isoleucine -9 threonine), 1409 (alanine -9 threonine),1624 (threonine -9 alanine), 1649 (arginine -9methionine), 1717 (aspartic acid -9 alanine), 1936(histidine -9 tyrosine), 2074 (glutamine -9 arginine)and 2114 (arginine -9 lysine).In the case of human parainfluenza virus type3, at least one attenuating mutation in the 3' genomicpromoter region is selected from the group consistingof nucleotide 23 (T -9 C), nucleotide 24 (C -9 T),nucleotide 28 (G -9 T) and nucleotide 45 (T -9 A), andat least one attenuating mutation in the RNA polymerasegene is selected from the group consisting ofnucleotide changes which produce changes in an aminoacid selected from the group consisting of residues 942(tyrosine -9 histidine), 992 (leucine -9SUBST?UTESHEET(RULE26)Y?ac\.\ux.kw\«u1L\cHL\ us0-’-.. H‘-M ../J- A‘.'-V1015202530-‘-In AL».11..phenylalanine). and 1558 (threonine -+ isoleucine).In the case of human respiratory syncytialvirus subgroup E, at least one attenuating mutation inthe 3' genomic promoter region is selected from thegroup consisting oi nucleotide 4 (C -9 G) and theinsertion of an additional A in the stretch of A’s atnucleotides 6-11, and at least one attenuating mutationin the RNA polymerase gene is selected fro the groupconsisting of nucleotide changes which produce changesin an amino acid selected from the group consisting ofresidues 353 (arginine -9 lysine), 451 (lysine -+arginine). 1229 (aspartic acid —+ a8PÂ¥r39i?9). 2029(threonine -9 isoleucine) and 2050 (asparagine -+aspartic acid).In another embodiment of this invention,attenuated virus is used to prepare vaccines whichelicit a protective immune response against the wild-type form of the virus.In yet another embodiment of this invention.an isolated. positive strand, antigenonic message sensenucleic acid molecule (or an isolated, negative strandgenomic sense nucleic acid molecule) having thecomplete viral nucleotide sequence (whether of wild-type virus or virus attenuated by non-recombinantmeans) is manipulated by introducing one or more of theattenuating mutations described in this application togenerate an isolated. recomhinantly-generatedattenuated virus. This virus is then used to preparevaccines which elicit a protective immune responseagainst the wild-type form of the virus.In still another embodiment or thisinvention. such a complete wi1d—type or vaccine viralnucleotide sequence is used: (1) to design PCR primersfor use in a PCR assay to detect the presence of theAMENDED SHEET'?3{K‘gQéAQ226sss4 1999-03-161~:~ fUUwuu— VIJ‘hJ ;HuHJ1nJ.d3}?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/16718corresponding virus in a sample; or (2) to design andselect peptides for use in an ELISA to detect thepresence of the corresponding virus in a sample.Brief Description Of The FiguresFigure 1 depicts the passage history of theEdmonston measles virus (15). The abbreviations havethe following meanings: HK - human kidney; HA - humanamnion; CE(am) — chick embryo; CEF - chick embryofibroblast; DK — dog kidney; WI-38 — human diploidcells; SK - sheep kidney; * — plaque cloning. Thenumber following each abbreviation represents thenumber of passages.Figure 2 depicts a map of the measles virusgenome showing putative cis—acting regulatory elementsat and near the genome and antigenome termini. Top - aschematic map of the measles virus genome, beginning atthe 3' end with 52 nucleotides of leader sequence (1)and ending at the 5' terminus with 37 nucleotides oftrailer sequence (t). Gene boundaries are denoted byvertical bars; below each gene is the number ofcistronic nucleotides. Bottom - an expanded schematicview of the 3' extended genomic promoter regions ofgenome and antigenome, showing the position andsequence of the two highly conserved domains, A and B.The intervening intergenic trinucleotide is denoted aswell. Nascent 5' RNAs encompassing the A‘ to B’regions are presumed to contain the regulatory sequenceat which the N protein encapsidation initiates.Figure 3 depicts a genetic map of the RSVsubgroup B wild-type strains designated 2B and 18537(top portion), the intergenic sequences of thosestrains (middle portion) and the 68 nucleotide overlapbetween the M2 and L genes (bottom portion). The RSVSUBST?UTESHEET(RULE26)I?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/167182B stain has six fewer nucleotides in the G gene,encoding two fewer amino acid residues in the GThe 2Bstrain has 145 nucleotides in the 5' trailer region, asprotein, as compared to the 18537 strain.compared to 149 nucleotides in the 18537 strain. The2B strain has one more nucleotide in each of the NS-1,NS-2 and N genes, and one fewer nucleotide in each ofthe M and F genes, as compared to the 18537 strain.Detailed Description Of The InventionTranscription and replication of negative-sense, single stranded RNA viral genomes are achievedthrough the enzymatic activity of a multimeric proteinacting on the ribonucleoprotein core (nucleocapsid).Naked genomic RNA cannot serve as a template. .Instead,these genomic sequences are recognized only when theyare entirely encapsidated by the N protein into thenucleocapsid structure. It is only in that contextthat the genomic and antigenomic terminal promotersequences are recognized to initiate thetranscriptional or replication pathways.All paramyxoviruses require the two viralproteins, L and P, for these polymerase pathways toproceed. The pneumoviruses, including RSV, alsorequire the transcription elongation factor, M2, forthe transcriptional pathway to proceed efficiently.Additional cofactors may also play a role, includingperhaps the virus—encoded N81 and NS2 proteins, as wellas perhaps host~cell encoded proteins.However, considerable evidence indicates thatit is the L protein which performs most, if not all,the enzymatic processes associated with transcriptionand replication, including initiation, and terminationof ribonucleotide polymerization, capping andSUBST?UTESHEET(RULE26)?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/ 16718polyadenylation of mRNA transcripts, methylation andperhaps specific phosphorylation of P proteins. The Lprotein's central role in genomic transcription andreplication is supported by its large size, sensitivityto mutations, and its catalytic level of abundance inthe transcriptionally active viral complex (16).These considerations led to the proposal thatL proteins consist of a linear array of domains whoseconcatenated structure integrates discrete functions(17). Indeed, three such delimited, discrete elementswithin the negative—sense virus L protein have beenidentified based on their relatedness to definedfunctional domains of other well-characterizedproteins. These include: (1) a putative RNA templaterecognition and/or phosphodiester bond formationdomain; (2) an RNA binding element; and (3) an ATPbinding domain. All prior studies of L proteins ofnonsegmented negative—sense, single stranded RNAviruses have revealed these putative functionalelements (17).Without being bound by the following, it isreasonable to presume that these non-protein coding,promoter and other cis-acting genomic regulatorydomains are important determinants of the efficiencywith which transcription and replication by measlesvirus (MV) and other viruses of the OrderMononegavirales are actualized, in association with theL protein, and that they may therefore be virulencedeterminants for these viruses as well.In summary, the invention is believed toencompass a coordinate set of changes between the cis-acting regulatory signal (3' genomic promoter region)and the polymerase gene (L) which results inattenuation of the virus while retaining sufficientability of the virus to replicate. Attenuation isSUBSTITUTE SHEET (RULE 25)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97Il67l8optimized by rational mutations of the 3' genomicpromoter region and the polymerase gene, which providethe desired balance of replication efficiency: so thatthe virus vaccine is no longer able to produce disease,yet retains its capacity to infect the vaccinee'scells, to express sufficiently abundant gene productsto elicit the full spectrum and profile of desirableimmune responses, and to reproduce and disseminatesufficiently to maximize the abundance of the immuneresponse elicited.Without being bound by the following,attenuating mutations in the extended promoter (3'genomic promoter region) and in the polymerase gene arebelieved to affect the display of cis-acting signalsand the conformation of the polymerase complex engagingthese signals. For example, when encapsidated, thepromoter RNA is coiled in a helical array. Changes inpromoter sequence may affect the relative positions atwhich the conserved signals are displayed relative toone another. Specifically, the measles wild—type 3'genomic promoter region has a pyrimidine (uracil) atpositions 26 and 42 (the antigenomic message sensesequences have the purine adenine). The vaccinestrains have purines at those positions (theantigenomic message sense sequences have thecorresponding pyrimidines; see Table 3 in Example 1below). The larger purines may change the distanceand/or angular display between the conserved domains ofthe promoter (e.g, in measles, positions 1-11 and 87-98). resulting in an altered spatial presentation ofthe cis-acting signals to the polymerase.Animal studies have demonstrated a decreasein viral replication sufficient to avoid illness butadequate to elicit the desired immune response. Thislikely represents a decrease in transcription, aSUBSTHUTESHEET(RULE26)?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/16718decrease in gene expression of virally encodedproteins, a decrease in antisense templates and,therefore, the production of fewer new genomes. Theresulting attenuated viruses are significantly lessvirulent than the wild-type.The attenuating mutations described hereinmay be introduced into viral strains by two methods:(1) Conventional means such as chemicalmutagenesis during virus growth in cell cultures towhich a chemical mutagen has been added, selection ofvirus that has been subjected to passage at suboptimaltemperature in order to select temperature sensitiveand/or cold adapted mutations, identification of mutantvirus that produce small plaques in cell culture, andpassage through heterologous hosts to select for hostrange mutations. These viruses are then screened forattenuation of their biological activity in an animalmodel. Attenuated viruses are subjected to nucleotidesequencing of their 3' genomic promoter region andpolymerase genes to locate the sites of attenuatingmutations. Once this has been done, method (2) is thencarried out.(2) A preferred means of introducingattenuating mutations comprises making predeterminedmutations using site-directed mutagenesis. Thesemutations are identified either by method (1) or byreference to closely-related viruses whose attenuatingmutations are already known. One or more mutations areintroduced into each of the 3' genomic promoter regionand the polymerase gene. Cumulative effects ofdifferent combinations of coding and non—coding changescan also be assessed.The mutations to the 3' genomic promoterregion and polymerase gene are introduced by standardrecombinant DNA methods into a DNA copy of the viralSUBSTHUTESHEET(RULE26)v?101520253035WO 98/13501CA 02265554 l999-03- 16PCT/US97/l 6718genome. This may be a wildutype or a modified viralgenome background (such as viruses modified by method(1)), thereby generating a new virus. Infectiousclones or particles containing these attenuatingmutations are generated using the cDNA "rescue" system,which has been applied to a variety of viruses,including Sendai virus (18); measles virus (19);respiratory syncytial virus (20); rabies (21);vesicular stomatitis virus (VSV) (15); and rinderpestvirus (23); these references are hereby incorporated byreference. See, for measles virus rescue, publishedInternational patent application WO 97/06270,designating the United States (24); for PIV—3 rescue,U.S. provisional patent application 60/047575 (25); forRSV rescue, published International patent applicationWO 97/12032, designating the United States (26); theseapplications are hereby incorporated by reference.Briefly, all Mononegavirales rescue systemscan be summarized as follows: Each requires a clonedDNA equivalent of the entire viral genome placedbetween a suitable DNA—dependent RNA polymerasepromoter (e.g., the T7 RNA polymerase promoter) and aself-cleaving ribozyme sequence (e.g., the hepatitisdelta ribozyme) which is inserted into a propagatablebacterial plasmid. This transcription vector providesthe readily manipulable DNA template from which the RNApolymerase (e.g., T7 RNA polymerase) can faithfullytranscribe a single-stranded RNA copy of the viralantigenome (or genome) with the precise, or nearlyprecise, 5’ and 3’ termini. The orientation of theviral genomic DNA copy and the flanking promoter andribozyme sequences determine whether antigenome orgenome RNA equivalents are transcribed. Also requiredfor rescue of new virus progeny are the virus-specifictrans-acting proteins needed to encapsidate the naked,SUBSTHUTESHEET(HULE26)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/U S97/ 16718single-stranded viral antigenome or genome RNAtranscripts into functional nucleocapsid templates:the viral nucleocapsid (N or NP) protein, thepolymerase-associated phosphoprotein (P) and thepolymerase (L) protein. These proteins comprise theactive viral RNA-dependent RNA polymerase which mustengage this nucleocapsid template to achievetranscription and replication.The trans-acting proteins required formeasles virus rescue are the encapsidating protein N,and the polymerase complex proteins, P and L. For PIV-3, the encapsidating protein is designated NP, and thepolymerase complex proteins are also referred to as Pand L. For RSV, the virus—specific trans-actingproteins include N, P and L, plus an additionalprotein, M2, the RSV—encoded transcription elongationfactor.Typically, these viral trans-acting proteinsare generated from one or more plasmid expressionvectors encoding the required proteins, although someor all of the required trans-acting proteins may beproduced within mammalian cells engineered to containand express these virus-specific genes and geneproducts as stable transformants.The typical (although not necessarilyexclusive) circumstances for rescue include anappropriate mammallian cell milieu in which T7polymerase is present to drive transcription of theantigenomic (or genomic) single-stranded RNA from theviral genomic cDNA-containing transcription vector.Either cotranscriptionally or shortly thereafter, thisviral antigenome (or genome) RNA transcript isencapsidated into functional templates by thenucleocapsid protein and engaged by the requiredpolymerase components produced concurrently from co-SUBSTHUTESHEET(RULE26)1?W0 98/1350]101520253035CA 02265554 l999-03- 16PCT/US97/1 6718-19-transfected expression plasmids encoding the requiredvirus-specific trans-acting proteins. These events andprocesses lead to the prerequisite transcription ofviral mRNAs, the replication and amplification of newgenomes and, thereby, the production of novel viralprogeny, i.e., rescue.For the rescue of rabies, VSV and Sendai, T7polymerase is provided by recombinant vaccinia virusVTF7-3 .rescued virus be separated from the vaccinia virus byThis system, however, requires that thephysical or biochemical means or by repeated passagingin cells or tissues that are not a good host forpoxvirus. For MV cDNA rescue, this requirement isavoided by creating a cell line that expresses T7polymerase, as well as viral N and P proteins. Rescueis achieved by transfecting the genome expressionvector and the L gene expression vector into the helpercell line. Advantages of the host—range mutant of thevaccinia virus, MVA—T7, which expresses the T7 RNApolymerase, but does not replicate in mammalian cells,are exploited to rescue RSV, Rinderpest virus and MV.After simultaneous expression of the necessaryencapsidating proteins, synthetic full lengthantigenomic viral RNA are encapsidated, replicated andtranscribed by viral polymerase proteins and replicatedgenomes are packaged into infectious virions. Inaddition to such antigenomes, genome analogs have nowbeen successfully rescued for Sendai and PIV-3 (25,27).The rescue system thus provides a compositionwhich comprises a transcription vector comprising anisolated nucleic acid molecule encoding a genome orantigenome of a nonsegmented, negative-sense, singlestranded RNA virus of the Order Mononegavirales havingat least one attenuating mutation in the 3' genomicpromoter region and having at least one attenuatingSUBSTHUTESHEET(RULE26).‘.W.,....‘.........-..........-......................._. . »?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718mutation in the RNA polymerase gene, together with atleast one expression vector which comprises at leastone isolated nucleic acid molecule encoding the trans-acting proteins necessary for encapsidation,transcription and replication (e.g., N, P and L formeasles virus; NP, P and L for PIV-3; N, P, L and M2for RSV). Host cells are then transformed ortransfected with the at least two expression vectorsjust described. The host cells are cultured underconditions which permit the co-expression of thesevectors so as to produce the infectious attenuatedvirus.The rescued infectious virus is then testedfor its desired phenotype (temperature sensitivity,cold adaptation, plaque morphology, and transcriptionand replication attenuation), first by in vitro means.The mutations at the cis-acting 31 genomic promoterregion are also tested using the minireplicon systemwhere the required trans-acting encapsidation andpolymerase activities are provided by wild-type orvaccine helper viruses, or by plasmids expressing theN, P and different L genes harboring gene-specificattenuating mutations (19,28).If the attenuated phenotype of the rescuedvirus is present, challenge experiments are conductedwith an appropriate animal model. Non-human primatesprovide the preferred animal model for the pathogenesisof human disease. These primates are first immunizedwith the attenuated, recombinantly—generated virus,then challenged with the wild-type form of the virus.Monkeys are infected by various routes, including butnot limited to intranasal, intratracheal orsubcutaneous routes of inoculation (29).Experimentally infected rhesus and cynomolgus macaqueshave also served as animal models for studies ofSUBSTHUTESHEET(RULE26)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718vaccine-induced protection against measles (30).Protection is measured by such criteria as diseasesigns and symptoms, survival, virus shedding andantibody titers. If the desired criteria are met, theattenuated, recombinantly-generated virus is considereda viable vaccine candidate for testing in humans. The"rescued" virus is considered to be "recombinantly-generated", as are the progeny and later generations ofthe virus, which also incorporate the attenuatingmutations.Even if a “rescued virus is underattenuatedor overattenuated relative to optimum levels forvaccine use, this is information which is valuable fordeveloping such optimum strains.optimally, a codon containing an attenuatingpoint mutation may be stabilized by introducing asecond or a second plus a third mutation in the codonwithout changing the amino acid encoded by the codonbearing only the attenuating point mutation.Infectious virus clones containing the attenuating andstabilizing mutations are also generated using the cDNA"rescue" system described above.Measles virus serves as a useful model forthis invention, because sequence data are now availableas described herein for the disease-causing wild—typevirus and for the disease—preventing vaccines whichhave a demonstrated history of efficacy.Measles virus was first isolated in tissueculture in 1954 (31) from an infected patient namedDavid Edmonston. This Edmonston strain of measlesbecame the progenitor for many live—attenuated measlesvaccines including Moraten, which is the currentvaccine in the United States (Attenuvaxm; Merck Sharp &Dohme, West Point, PA) and was licensed in 1968 and hasproven to be efficacious.SUBST?UTESHEET(RULE26)?101520253035WO 98/13501CA 02265554 l999-03- 16PCT /US97l16718Aggressive immunization programs institutedin the mid to late 1960s resulted in the precipitousdrop in reported measles cases from near 700,000 in1965 to 1500 in 1983. In parallel, other vaccinestrains were also developed from the Edmonston strain(see Fig. 1), Schwarz (Institut Merieux, Lyon, France),Zagreb (Zagreb, Yugoslavia) and AIK—C (Japan). Theseother vaccines have also proven to be efficacious andhave been used extensively. An early, reactogenic,underattenuated vaccine strain (Rubeovaxmz Merck Sharp& Dohme) produced measles-like illness in children andits use thus was discontinued. It, however, wasfurther attenuated successfully to produce the Moratenvaccine strain (see Fig. 1) (32). Live measles virusvaccine provides a success story of the development ofan efficacious vaccine and provides a model forunderstanding the molecular mechanisms of viral vaccineattenuation among nonsegmented, negative-sense, singlestranded RNA viruses.Because of its significance as a major causeof human morbidity and mortality, measles virus (MV)has been quite extensively studied. MV is a large,relatively spherical, enveloped particle composed oftwo compartments, a lipoprotein membrane and aribonucleoprotein particle core, each having distinctbiological functions (33). The virion envelope is ahost cell—derived plasma membrane modified by threevirus—specified proteins: The hemagglutinin (H;approximately 80 kilodaltons (kD)) and fusion (FL2;approximately 60 kD) glycoproteins project on thevirion surface and confer host cell attachment andentry capacities to the viral particle (16).Antibodies to H and/or F are considered protectivesince they neutralize the virus’ ability to initiateinfection (34,35,36). The matrix (M; approximately 37SUBSTHUTESHEET(RULE26)I?101520253035WO 98113501CA 02265554 l999-03- 16PCT/US97/16718kD) protein is the amphipathic protein lining themembrane’s inner surface, which is thought toorchestrate virion morphogenesis and thus consummatevirus reproduction (37). The virion core contains the15,894 nucleotide long genomic RNA upon which templateactivity is conferred by its intimate association withapproximately 2600 molecules of the approximately 60 kDnucleocapsid (N) protein (38,39,40). Looselyassociated with this approximately one micron longhelical ribonucleoprotein particle are enzymatic levelsof the viral RNA dependent RNA polymerase (L;approximately 240 kD) which in concert with thepolymerase cofactor (P; approximately 70 kD), andperhaps yet other virus—specified as well ashost-encoded proteins, transcribes and replicates theMV genome sequences (41).To date, the entire nucleotide sequences(only for the Edmonston B laboratory strain and theAIK-C vaccine strain), coding potential, andorganization of the MV genome have been reported (33).The six virion structural proteins are encoded by sixcontiguous, non-overlapping genes which are arrayed as3 '—N—P-M-—F-H-L—5 ' .products of as yet uncertain function have also beenfollows: Two additional MV geneidentified. These two nonstructural proteins, known asC (approximately 20 kD) and V (approximately 45 kD),are both encoded by the P gene, the former by a secondreading frame within the P mRNA; the latter by acotranscriptionally edited P gene—derived mRNA whichencodes a hybrid protein having the amino terminalsequences of P and a new zinc finger-like cysteine—richcarboxy terminal domain (16).In addition to the sequences encoding thevirus—specified proteins, the MV genome containsdistinctive non—protein coding domains resembling thoseSUBSTITUTE SHEEET (RULE 26)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/U S97/ 16718directing the transcriptional and replicative pathwaysof related viruses (16,42). These regulatory signalslie at the 3' and 5' ends of the MV genome and in shortinternal regions spanning each intercistronic boundary.The former encode the putative promoter and/orregulatory sequence elements directing genomictranscription, genome and antigenome encapsidation, andreplication. The latter signal transcriptiontermination and polyadenylation of each monocistronicviral mRNA and then reinitiation of transcription ofthe next gene. In general, the MV polymerase complexappears to respond to these signals much as theRNA-dependent RNA polymerases of other non—segmentednegative strand RNA viruses (16,42,43,44).Transcription initiates at or near the 3' endof the MV genome and then proceeds in a 5' directionproducing monocistronic mRNAs (40,42,45). As thepolymerase traverses the MV genomic template, itencounters putative stop/start signals which, in 3' to5' order, are: a semi-conserved transcriptiontermination/polyadenylation signal (A/G U/C UA A/U NNA4, where N may be any of the four bases) at which eachmonocistronic RNA is completed; a non-transcribedintergenic trinucleotide punctuation mark (CUU; exceptat the H:L boundary where it is CGU); and asemiconserved start signal for transcription initiationof the next gene (AGG A/G NN C/A A A/G G A/U, where Nmay be any of the four bases) (45,46). Since somepolymerase complexes fail to reinitiate, the abundanceof each MV mRNA diminishes in parallel with thedistance of the encoding gene from the genomic 3' end.This mRNA gradient directly corresponds to the relativeabundance of each virus-specified protein. Thisindicates that MV protein expression is ultimatelycontrolled at the transcriptional level (44).SUBSTITUTE SHEET (RULE 26)I?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/U S97/ 16718-25-The 3' and 5' MV genomic termini containnon—protein coding sequences with distinct parallels tothe leader and trailer RNA encoding regions of VSV(42). Nucleotides 1-55 define the region between thegenomic 3' terminus and the beginning of the N gene,while 37 additional nucleotides can be found betweenthe end of the L gene and the 5' terminus of thegenome. However, unlike VSV, or even theparamyxoviruses Sendai and NDV, MV does not transcribethese terminal regions into short, unmodified (+) or(-) sense leader RNAs (47,48,49). Instead, leaderreadthrough transcripts, including full—lengthpolyadenylated leader:N, leader:N:P, leader:N:P:M, andof course full-length antigenome MV RNAs aretranscribed (48,49). Thus, the short leadertranscript, the key operational element determining theswitch from transcription to replication of the VSVsingle—stranded, negative polarity genome (50,5l,52),seems absent in MV. This leads to consideration andexploration of alternative models for this crucialreproductive event (42).Measles virus, as well as all otherMononegavirales except the rhabdoviruses, appears tohave extended its terminal regulatory domains beyondthe confines of leader and trailer encoding sequences(42). For measles, these regions encompass the 107 3'genomic nucleotides (the "3' genomic promoter region",also referred to as the "extended promoter", whichcomprises 52 nucleotides encoding the leader region,followed by three intergenic nucleotides, and 52nucleotides encoding the 5' untranslated region of NmRNA) and the 109 5' end nucleotides (69 encoding the3' untranslated region of L mRNA, the intergenictrinucleotide and 37 nucleotides encoding the trailer).Within these 3' terminal approximately 100 nucleotidesSUBSTHUTESHEET(RULE26)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718of both the genome and antigenome are two short regionsof shared nucleotide sequence: 14 of 16 nucleotides atthe absolute 3' ends of the genome and antigenome areidentical. Internal to those termini, an additionalregion of 12 nucleotides of absolute sequence identityhave been located. Their position at and near thesites at which the transcription of the MV genome mustinitiate and replication of the antigenome must begin,suggests that these short unique sequence domainsencompass an extended promoter region.These discrete sequence elements may dictatealternative sites of transcription initiation —- theinternal domain mandating transcription initiation atthe N gene start site, and the 3' terminal domaindirecting antigenome production (42,48,53). Inaddition to their regulatory role as cis-actingdeterminants of transcription and replication, these 3'extended genomic and antigenomic promoter regionsencode the nascent 5' ends of antigenome and genomeRNAs, respectively. Within these nascent RNAs resideas yet unidentified signals for N protein nucleation,another key regulatory element required fornucleocapsid template formation and consequently foramplification of transcription and replication. Figure2 schematically shows the location and sequence ofthese highly conserved, putative cis-acting regulatorydomains.Terminal non—protein coding regions similarin location, size and spacing are present in thegenomes of other members of the genus Paramyxoviridae,though only 8-11 of their absolute terminal nucleotidesare shared by MV (42,54). The genomic terminii of theMorbillivirus canine distemper virus (CDV) displays agreater degree of homology with its MV relative: 73%of the nucleotides of the leader and trailer sequencesSUBSTITUTE SHEET (RULE 26)1?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718-2']-of these two viruses are identical, including 16 of 18at the absolute 3' termini and 17 of 18 at their 5'ends (55). No accessory internal CDV genomic domain-sharing homology to that of the MV extended promoterhas been found. However, there is a 20 nucleotide longstretch lying between CDV genomic nucleotides 85 and104 and 15,587 and 15,606 in which 15 of the 20nucleotides are complementary (Gene Bank accessionnumber AF 14953). This indicates that CDV, like MVcontains an additional region within its non—coding 3'genomic and antigenomic ends that may provide importantcis-acting promoter and/or regulatory signals (55).Additionally, the precise length of the 3'-leader region (55 nucleotides) is identical amongseveral members of the Family Paramyxoviridae (MV, CDV,PIV-3, BPV—3, SV and NDV). Further evidence for theimportance of these extended, non-protein codingregions comes from analyses of a large number ofdistinct copy-back Defective Interfering Viruses (DIs)recently cloned from subacute sclerosingpanencephalitis (SSPE) brain tissue. No DI with a stemshorter than the 95 5' terminal genomic nucleotides wasfound. This indicates that the minimal signals neededfor MV DI RNA replication and encapsidation extend wellbeyond the 37 nucleotide long trailer sequence toencompass the additional internal putative regulatorydomain (56).As exemplified in part by measles virus, thisinvention is directed to the concept that importantvirulence/attenuation determinants reside in viralgenomic non-protein coding regulatory regions and inthe transacting transcription/replication enzymecomplex with which these cis-acting elements mustinteract. The cis-acting domains are found both at the3' and 5' ends of the MV gename, flanking the sixSUBSTHUTESHEETKRULEZG)?1015202530W0 98/ 13501CA 02265554 l999-03- 16PCT/U S97/ 16718contiguous genes encoding viral structural proteins;and within the MV genome as short regions encompassinginternal intergenic boundaries. The former encode theputative promoter and/or regulatory sequence elementsdirecting the vital processes of genomic transcription,genome and antigenome encapsidation, and replication.The latter signal transcription termination andpolyadenylation of each monocistronic viral mRNA andthen reinitiation of transcription of the next gene.The transcription/replication enzyme, RNA dependent RNApolymerase molecule can modulate transcription and/orreplicative efficiency, thereby determining theabundance of cytopathic viral gene products and/orvirion progeny.Proof of the concept of this invention formeasles virus is obtained by first determining thenucleotide sequences of the non—coding regulatoryregions (3' genomic promoter region) and the codingregions of the L gene (with predicted amino acidsequences) of the progenitor Edmonston wild-type MVisolate, together with available measles vaccinestrains derived from this isolate (see Figure 1).Independent other wild-type isolates were examined forcomparative purposes as well.The nucleotide sequences (in positive strand,antigenomic, message sense) of four wild-type and fivevaccine measles strains, as well as the deduced aminoacid sequences of the RNA polymerase (L protein) ofthese measles viruses, are set forth as follows withreference to the appropriate SEQ ID NOS. containedherein:SUBSTITUTE SHEET (RULE 26)I?101520253035........,..w.........«.......,.........-W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/16718_ 29 _Virus Nucleotide Segyence L Protein SeggenceWild-TypeEdmonston SEQ ID NO:1 SEQ ID NO:21977 SEQ ID NO:3 SEQ ID NO:41983 SEQ ID NO:5 SEQ ID NO:6Montefiore SEQ ID NO:7 SEQ ID NO:8Vaccine _Rubeovax” SEQ ID NO:9 SEQ ID NO:10Moraten SEQ ID NO:ll SEQ ID NO:l2Zagreb SEQ ID N0:13 SEQ ID NO:14AIK-C SEQ ID NO:15 SEQ ID NO:l6Each measles virus genome listed above is15,894 nucleotides in length. Translation of the Lgene starts with the codon at nucleotides 9234-9236;the translation stop codon is at nucleotides 15783-15785. The translated L protein is 2,183 amino acidslong.Note that nucleotide 2499 of 1983 wild-typemeasles virus is indicated as “G” in SEQ ID NO:5. Infact, the base is actually a mixture of “G” and “C”.Also note that nucleotide 2143 of Rubeovax” vaccinevirus is indicated as “T” in SEQ ID NO:9. In nineclones sequenced, this base was “T” in seven and “C” intwo; thus, this base can be “T” or “C”.In addition, the Schwarz vaccine virus genomeis identical to that of the Moraten vaccine virusgenome (SEQ ID NO:11), except that at nucleotides 4917and 4924, Schwarz has a “C” instead of a “T”.Nucleotide differences distinguishing the 3'genomic promoter region and nucleotide and amino aciddifferences distinguishing the L gene and L proteinsequences of the Edmonston wild—type isolate, vaccinestrains and other independently isolated wild—typeSUBSTITUTE SHEET (RULE 26)?101520253035W0 98/13501ICA 02265554 l999-03- 16PCT/US97/16718viruses were then compared and aligned (see Tables 3-5in Example 1 below).As shown in Table 3, there were threemutations from the 3' genomic promoter region (inantigenomic, message sense) of the progenitor wild-typeMV isolate and the derivative vaccine strains: Atnucleotide position 26, from "A" to "T"; at position42, from "A" to "C" or from “A” to "T"; and in the caseof Zagreb only, at position 96, from "G" to "A". Inaddition, the other examined wild-type isolatesdiffered from both the progenitor wild-type isolate andthe vaccine strains at position 50 by having "A"instead of "G".The predicted amino acid sequences of the Lgenes of measles vaccine strains (Rubeovaxm, Moraten,Schwarz, AIK—C and Zagreb) and wild-type isolates(1977, 1983 and Montefiore), differ from the progenitorstrain (Edmonston) at 49 positions in the 2183 aminoacid long open reading frame (see Tables 4 and 5 inExample 1 below).These amino acid differences can be dividedinto four categories:(1) Positions where one vaccine straindiffers from the progenitor, as well as from othervaccine and wild-type strains, suggesting a potentialattenuation site.(2) Specific differences between all wild-type and all vaccine sequences; these may alsoconstitute important attenuation sites.3) Residues where chronologically newer wild-types differ from older wild—types; which may beattributable to genetic drift.(4) Positions where one or more vaccinestrains and/or wild-type strains have common aminoacids and differ from all the other strains; theseSUBSTITUTE SHEET (RULE 26)I?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/16718changes may represent lineage—specific, potentiallyattenuating changes within the vaccine strains andrelatedness among the wild-type isolates, respectively.There were four category (1) changes whereone vaccine differed from the other vaccines, as wellas the wild-type strains. Two of these were in Moratenand Schwarz (amino acids 331 and 2114) and two were inAIK—C (1624 and 2074). These mutations are of specialinterest because all of these viruses are goodvaccines. Thus, these positions are sites forattenuation.Only one position, 1717, fits into category(2), with all wild—types having aspartic acid and allvaccines having alanine. Interestingly, this positionis in one of two areas where the L genes of measles andcanine distemper virus (which are otherwise highlyhomologous) do not show exceptional conservation. Thisdifference makes it more likely that 1717 is a keyposition for an attenuating mutation in measles.There were five positions, 149, 636, 720,2017 and 2119, where both chronologically newer wild-types (1983 and Montefiore) differ from older wild-types (Edmonston and 1977), which therefore fit intocategory (3). These differences suggest genetic driftrather than denoting sites of attenuating mutations.Not included in this total are 16 positions whereMontefiore (the 1989 isolate) differed from the rest(see Table 5). These could be either genetic drift(category (3)) or random change (category (4)). Theremaining 23 positions are category (4), with one ormore of the viruses differing from the consensus.Three of these positions (1409, 1649, 1936)are potentially attenuating category (4) mutations.These are changes where two vaccine strains have acommon change from the progenitor wild-type strain.SUBSTHUTESHEET(RULE26). .,._.....—..—.._....._—_-..-.-..n.—................ V?101520253035WO 98/13501CA 02265554 l999-03- 16PCT/US97/16718These changes may be connected with the vaccine lineageleading to the Rubeovax” and Moraten vaccines (Figure1).Applicants have found that their AIK-Cvaccine strain nucleotide sequence differs from thepublished sequence (33) at 21 positions, including oneinsertion and one deletion. Several of thesedifferences result in coding changes including two inthe L gene (at amino acids 1477 and 2008).Thus, the additional changes accrued withinthe L gene sequence as the measles progenitor strain isprogressively attenuated to achieve a replicativecapacity optimized for live vaccine purposes appears tobe constrained and delimited. Presumably, this limitedtolerance in the number and location of L gene changesis imposed not only by the need to preserve themultifunctional capacities of the polymerase, but alsoby the preexisting 3’ promoter changes with which theevolving L protein must interact to achievetranscription and replication. In other words, optimalvirus attenuation requires coordinate (i.e., linked)changes in the polymerase protein and the cis-actingregulatory elements on which it acts.The 3'—leader displays the least tolerancefor change, allowing highly selected changes during theattenuation process at nucleotide position 26 (alwaysthe change of from "A" to "T"), and at position 42 (thechange of from "A" to "C" or from “A” to "T") (inantigenomic, message sense). In the case of Zagrebonly, there is a single further change, from "G" to “A”at position 96, which may be important when combinedwith Zagreb L gene—specific changes. The 3'—leaderregion seems to have undergone only one instance ofgenetic drift since 1954, with a change of "G" to "A"at position 50 (see Table 3).SUBSTHUTESHEET(RULE25)I?CA 02265554 l999-03- 16W0 98/ 13501 PCT/U S97/ 16718The net change in the 3' genomic promoterregion during the attenuation process is thereplacement of two pyrimidines by two purines ingenomic sense in all MV vaccine strains. The co-5 evolution of the L gene during these attenuationprocesses is believed to reflect selection of subtlechanges favoring reproduction of the viruses indifferent host cells. All the vaccine strains weregrown in chick embryo (CE) or chick embryo fibroblast10 (CEF) cells during their attenuation process (Figure1). In addition, some vaccine strains have beenexposed to unique host cells; i.e., Zagreb vaccine wasgrown in dog kidney cells and human diploid cells,while the AIK—C vaccine was adapted to sheep kidney15 cells. Moraten and Rubeovax“ were exclusivelydeveloped in CE and CEF.Some of the lineage—specific L gene changes(position 1649 in Rubeovax”, Moraten and Schwarzvaccines and the change at position 1717 in all20 vaccines) represent a subset of adaptations of the Lgene to the 3'-leader to modulate thetranscription/replication processes for vaccineattenuation. Additionally, individual vaccine—specificchanges (category (1)) may provide additional fine tune25 modulation of virus replication/transcription for eachvaccine strain.Based on Table 3 and the foregoingdiscussion, the key attenuating mutations for the MV 3'genomic promoter region are nucleotide 26 (A -9 T),30 nucleotide 42 (A ->T or A —>C) and nucleotide 96 (G —>A) (in antigenomic, message sense).Based on Table 4 and the foregoingdiscussion, the key attenuating sites for the L proteinare as follows: amino acid residues 331 (isoleucine -935 threonine), 1409 (alanine —>threonine), 1624SUBSTHUTESHEET(RULE26)?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/U S97/ 16718(threonine —+alanine), 1649 (arginine —> methionine),1717 (aspartic acid —>alanine), 1936 (histidine -9tyrosine), 2074 (glutamine ~>arginine) and 2114(arginine —>lysine). It is understood that thenucleotide changes responsible for these amino acidchanges are not limited to those set forth in Table 4of Example 1 below; all changes in nucleotides whichresult in codons which are translated into these aminoacids are within the scope of this invention.Human parainfluenza virus type 3 (HPIV—3) isanother nonsegmented, negative-sense, single strandedenveloped RNA virus. HPIV-3 belongs to the FamilyParamyxoviridae (see Table 1). The genome of HPIV-3 is15,462 nucleotides long and encodes six non—overlappingprotein-encoding genes (57). Five of the genes encodea single virion structural protein each, which aredesignated NP (corresponding to the N protein of MV),M, F, HN (hemagglutinin—neuraminidase) and L. Thesixth mRNA encodes the P protein, and by an overlapping5' proximal open reading frame (ORF) encodes the Cprotein, and by the RNA editing mechanism, also encodesthe D protein.Like MV, HPIV-3 consists of a 3'—nonproteincoding leader region of 55 nucleotides, but unlikemeasles (where it is 37 nucleotides), it has a 44nucleotide long 5'—trailer region. The polymerasetranscribes the genome in a linear, sequential, start-stop manner which is guided by transcription signals inthe RNA template.Attempts to develop a live attenuated HPIV-3vaccine by passaging the wild-type virus JS strainthrough cell culture at sub-optimal temperature hasproduced promising results (7.57). Several "coldpassage" (cp) mutants were isolated for evaluation fromdifferent passage levels of the JS strain. One suchSUBSHTUTESHEET(RULE26)9?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/ 16718-35-mutant resulted from 45 serial passages and wasdesignated cp45.This virus exhibited three interestingproperties: (1) cold adaptation (ca): the ability toreplicate efficiently at the suboptimal temperature of20°C; (2) temperature sensitivity (ts): inability toreplicate in vitro at temperatures greater than orequal to 39°C; and (3) small plaque morphology. Thismutant appeared to be a promising vaccine candidatebecause: (a) its ca, ts and small plaque phenotype isstable after passage in cell culture; (b) itsreplication is restricted in both the upper and lowerrespiratory tract of hamsters; and (c) it inducedsignificant protection in hamsters against subsequentchallenge with wild-type HPIV-3 (58,59).Evaluation of this strain in the rhesusmonkey showed the attenuation mutations in cp45 to be acombination of ts and non-ts mutations (60).Subsequent evaluation in chimpanzees indicated thatcp45 appeared to be satisfactorily attenuated whilestill able to induce a high level of protection againstwild-type virus challenge (61). Later preliminaryclinical evaluation of cp45 in seronegative humaninfants and small children suggested that thiscandidate vaccine strain is suitably infectious andattenuated, as well as being moderately immunogenic(61).The cp45 strain has been grown in both fetalrhesus lung (FRhL) and Vero cells as follows: The PIV-3 cp45 virus grown in FRhL cells was prepared byinoculating confluent FRhL cell monolayers in tissueculture flasks at an MOI 0.1~1Ø The infected cellcultures were fed with EMEM medium and incubated at32°C. About seven days later, when maximal cytopathiceffects (synctyia) were observed, the virus wasSUBSTITUTE SHEET (RULE 26)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718harvested by subjecting the cultures to one freeze-thawcycle, pooling the fluids and then storing the virus at—70°C.The PIV-3 cp45 virus grown in Vero cells wasprepared by inoculating with virus a bioreactor cultureof confluent monolayers of Vero cells on microcarrierbeads which was continuously stirred. The infectedbioreactor culture was maintained at 30°C. The viruswas harvested 4-5 days later when syncytial CPE wasobserved. The culture fluid containing the virus wasstored at -70°C.The nucleotide sequences (in positive strand,antigenomic, message sense) of the HPIV—3 JS wild-typestrain (89) and the cp45 vaccine strain grown in FRhLand Vero cells, as well as the deduced amino acidsequences of the RNA polymerase (L protein) of theseHPIV—3 viruses, are set forth as follows with referenceto the appropriate SEQ ID NOS. contained herein:Virus Nucleotide Seggence L Protein SequenceWild-TypeJS SEQ ID NO:17 SEQ ID NO:l8VaccineFRhL cp45 SEQ ID NO:l9 SEQ ID NO:20Vero cp45 SEQ ID NO:2l SEQ ID NO:22Each PIV-3 virus genome listed above is15,462 nucleotides in length. Translation of the Lgene starts with the codon at nucleotides 8646-8648;the translation stop codon is at nucleotides 15345-15347. The translated L protein is 2,233 amino acidslong.As detailed in Example 2 and Table 6 thereinbelow, based upon the differences between the wild-typeSUBSTITUTE SHEET (RULE 26)1?s.‘..;A L.\._L‘A~4 .1..,;n_,Ru. \0.\ :H.\_\“ ,.;_\L-ML.‘-‘\ Ur, ;g7—11-.CA 02265554 1999-03—1(,lt;:ts T'.J:m:m- ++:,- am ‘_‘:v.;:H«-m.'3.;4 4.-.nu> .»4.. .I.-\.. . ._ -._as strain and the FRhL-grown op 45 mutant vaccinestrain, the key attenuating mutations tor the HPIV-3 3'genomic promoter region are nucleotide 23 (T —>c),nucleotide 24 (C -9T), nucleotide 28 (G -+T) and55 nucleotide 45 (T —>A) (in antigenomio. message sense).he also detailed in Example 2 and Table 6 thereinbelow, key attenuating sites for the L protein o£ HPIV-3 include the following: amino acid residues 942(tyrosine —>histidine), 992 (leucine —+ phenylalanine)_, 10 and 1558 (threonine -aisoleucine).N. It is understood that the nucleotide changesresponsible for these amino acid changes are notlimited to those set forth in Example 2 below; allchanges in nucleotides which result in oodons which are15 translated into these amino acids are within the scopeo£ this invention.Human respiratory syncytial virus (RSV) isyet another nonsegmented; negative-sense. single _stranded enveloped RNA virus. RSV belongs to the20 subfamily Pneumovirinae and the genus Pneumovirus (seeTable 1).Two major subgroups of human RSV, designatedA and B, have been identified based on reactivities oithe F and G eur?ace glycoproteins with monoclonal25 antibodies (62). More recently. the A and B lineagesoi RSV strains have been confirmed by sequence analysis(63.64). Bovine. ovine, and oaprine strains of thisvirus have also been isolated. The host speciticity ofthe virus it most clearly associated with the G30 attachment protein, which is highly divergent betweenAMENDED SHEET?1015202530W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718the human and the bovine/ovine strains (65,66), and maybe influenced, at least in part, by receptor binding.RSV is the primary cause of serious viralpneumonia and bronchiolitis in infants and youngchildren. Serious disease, i.e., lower respiratorytract disease (LRD), is most prevalent in infants lessthan six months of age. It most commonly occurs in thenonimmune infant's first exposure to RSV. RSVadditionally is associated with asthma andhyperreactive airways and it is a significant cause ofmortality in “high risk" children with bronchopulmonarydysplasia and congenital heart disease (CHD). It isalso one of the common viral respiratory infectionspredisposing to otitis media in children. In adults,RSV generally presents as uncomplicated upperrespiratory illness; however, in the elderly it rivalsinfluenza as a predisposing factor in the developmentof serious LRD, particularly bacterial bronchitis andpneumonia. Disease is always confined to therespiratory tract, except in the severelyimmunocompromised, where dissemination to other organscan occur. Virus is spread to others by fomitescontaminated with virus-containing respiratorysecretions, and infection initiates through the nasal,oral, or conjunctival mucosa.RSV disease is seasonal and virus is usuallyisolated only in the winter months, e.g., from Novemberto April in northern latitudes. The virus isubiquitous, and over 90% of children have been infectedat least once by 2 years of age. Multiple strainscocirculate. There is no direct evidence of antigenicdrift (such as that seen with influenza A viruses), butsequence studies demonstrating accumulation of aminoacid changes in the hypervariable regions of the GSUBSTHUTESHEET(RULE25)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718protein and SH proteins suggest that immune pressuremay drive virus evolution.In mouse and cotton rat models, both the Fand G proteins of RSV elicit neutralizing antibodiesand immunization with these proteins alone provideslongterm protection against reinfection (67,68).In humans, complete immunity to RSV does notdevelop and reinfections occur throughout life (69,70);however, there is evidence that immune factors willprotect against severe disease. A decrease in severityof disease is associated with two or more priorinfections and there is evidence that children infectedwith one of the two major RSV subgroups may be somewhatprotected against reinfection with the homologoussubgroup (71), observations which suggest that a liveattenuated virus vaccine may provide protectionsufficient to prevent serious morbidity and mortality.Infection with RSV elicits both antibody and cellmediated immunity. Serum neutralizing antibody to theF and G proteins has been associated, in some studies,with protection from LRD, although reduction in upperrespiratory disease (URD) has not been demonstrated.High levels of serum antibody in infants is associatedwith protection against LRD, and adminstration ofintravenous immunoglobulin with high RSV neutralizingantibody titers has been shown to protect againstsevere disease in high risk children (70,72,73). Therole of local immunity, and nasal antibody inparticular, is being investigated.The RSV virion consists of aribonucleoprotein core contained within a lipoproteinenvelope. The virions of pneumoviruses are similar insize and shape to those of all other paramyxoviruses.When visualized by negative staining and electronmicroscopy, virions are irregular in shape and range inSUBSTHUTESHEET(RULE26)?101520253035WO 98/13501CA 02265554 l999-03- 16PCT/U S97/ 16718diameter from 150-300 nm (74). The nucleocapsid ofthis virus is a symmetrical helix similar to that ofother paramyxoviruses, except that the helical diameteris 12-15 nm rather than l8nm. The envelope consists ofa lipid bilayer that is derived from the host membraneand contains virally coded transmembrane surfaceglycoproteins. The viral glycoproteins mediateattachment and penetration and are organized separatelyinto virion spikes. All members of paramyxovirussubfamily have hemagglutinating activity, but thisfunction is not a defining feature for pneumoviruses,being absent in RSV but present in PVM (75).Neuraminidase activity is present in members of thegenera Paramyxovirus, Rubulavirus, and is absent inMorbillivirus and Pneumovirus of mice (PVM) (75).RSV possesses two subgroups, designated A andB. The wild-type RSV (strain 2B) genome is a singlestrand of negative—sense RNA of 15,218 nucleotides (SEQID NO:23) that are transcribed into ten majorsubgenomic mRNAs. Each of the ten mRNAs encodes amajor polypeptide chain: Three are transmembranesurface proteins (G, F and SH); three are the proteinsassociated with genomic RNA to form the viralnucleocapsid (N, P and L); two are nonstructuralproteins (NS1 and NS2) which accumulate in the infectedcells but are also present in the virion in traceamounts and may play a role in regulating transcriptionand replication; one is the nonglycosylated virionmatrix protein (M); and the last is M2, anothernonglycosylated protein recently shown to be an RSV-specified transcription elongation factor (see Figure3). These ten viral proteins account for nearly all ofthe viral coding capacity.The viral genome is encapsidated with themajor nucleocapsid protein (N), and is associated withSUBSTITUTE SHEET (RULE 26)1?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/16718-41..the phosphoprotein (P), and the large (L) polymeraseprotein. These three proteins have been shown to benecessary and sufficient for directing RNA replicationof cDNA encoded RSV minigenomes (76). Further studieshave shown that for transcription to proceed with fullprocessing, the M2 protein (ORF 1) is required (74).When the M2 protein is missing, truncated transcriptspredominate, and rescue of the full length genome doesnot occur (74).Both the M (matrix protein) and the M2proteins are internal virion—associated proteins thatare not present in the nucleocapsid structure. Byanalogy with other nonsegmented negative-stranded RNAviruses, the M protein is thought to render thenucleocapsid transcriptionally inactive beforepackaging and to mediate its association with the viralenvelope. The NS1 and NS2 proteins have only beendetected in very small amounts in purified virions, andTheirfunctions are uncertain, though they may be regulatorsat this time are considered non-structural.of transcription and replication. Three transmembranesurface glycoproteins are present in virions: G, F, andSH. G and F (fusion) are envelope glycoproteins thatare known to mediate attachment and penetration of thevirus into the host cell. In addition, theseglycoproteins represent major independent immunogens(77).although a recent report has implicated its involvementThe function of the SH protein is unknown,in the fusion function of the virus (78).The genomes of two wild—type RSV subgroup Bstrains (2B and 18537) have now been sequenced in theirentirety (see SEQ ID NOS:23 and 25, discussed below).Genomic RNA is neither capped nor polyadenylated (79).In both the virion and intracellularly, genomic RNA istightly associated with the N protein.SUBSTITUTE SHEET (RULE 26)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718The 3' end of the genomic RNA consists of a44—nucleotide extragenic leader region that is presumedto contain the major viral promoter (Fig. 3). The 3'genomic promoter region is followed by ten viral genesin the order 3'—NSl-NS2—N-P—M-SH—G—F-M2-L-5' (Fig. 3).The L gene is followed by a 145-149 nucleotideextragenic trailer region (see Figure 3). Each genebegins with a conserved nine-nucleotide gene startsignal 3'-GGGGCAAAU (except for the ten-nucleotide genestart signal of the L gene, which is 3'-GGGACAAAAU;differences underlined). For each gene, transcriptionbegins at the first nucleotide of the signal. Eachgene terminates with a semi-conserved 12-14 nucleotidegene end (3'-A G U/G U/A ANNN U/A Abs) (where N can beany of the four bases) that directs transcriptiontermination and polyadenylation (Fig. 3). The firstnine genes are non-overlapping and are separated byintergenic regions that range in size from 3 to 56nucleotides for RSV B strains (Fig. 3). The intergenicregions do not contain any conserved motifs or anyobvious features of secondary structure and have beenshown to have no influence on the preceding andsucceeding gene expression in a minreplicon system(Fig. 3). The last two RSV genes overlap by 68nucleotides (Fig. 3). The gene-start signal of the Lgene is located inside of, rather than after, the M2gene. This 68 nucleotide overlap sequence encodes thelast 68 nucleotides of the M2 mRNA (exclusive of thePoly—A tail), as well as the first 68 nucleotides ofthe L mRNA.Ten different species of subgenomicpolyadenylated mRNAs and a number of polycistronicpolyadenylated read—through transcripts are theproducts of genomic transcription (74).Transcriptional mapping studies using UV light mediatedSUBSTITUTE SHEET (RULE 26)?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/16718-43..genomic inactivation showed that RSV genes aretranscribed in their 3' to 5' order from a singlepromoter near the 3' end (80). Thus, RSV synthesisappears to follow the single entry, sequentialtranscription model proposed for all Mononegavirales(16,81). According to this model, the polymerase (L)contacts genomic RNA in the nucleocapsid form at the 3'genomic promoter region and begins transcription at thefirst nucleotide. RSV mRNAs are co-linear copies ofthe genes, with no evidence of mRNA editing orsplicing.Sequence analysis of intracellular RSV mRNAsshowed that synthesis of each transcript begins at thefirst nucleotide of the gene start signal (74). The 5'end of the mRNAs are capped with the structurem7G(5')ppp(5')§p (where the underlined G is the firsttemplate nucleotide of the mRNA) and the mRNAs areBoth of thesemodifications are thought to be made co-polyadenylated at their 3' ends (82).transcriptionally by the viral polymerase. Threeregions of the RSV 3' genomic promoter have been foundTheseregions are the first ten nucleotides (presumablyto be important as cis acting elements (83).acting as a promoter), nucleotides 21-25, and the geneUnlikeother Paramyxovirinae, such as measles, Sendai and PIV-start signal located at nucleotides 45-53 (83).3. the remainder of the leader and non-coding region ofNS1 gene of RSV was found to be highly tolerant ofinsertions, deletions and substitutions (83).Additionally, by saturation mutagenesis(wherein each base is replaced independently by each ofthe other three bases and compared for translation andreplication efficiencies) within the first 12nucleotides of the 3' genomic promoter region, a U-tract located at nucleotides 6-10 was shown to beSUBSTITUTE SHEET (RULE 26)?101520253035W0 98/13501CA 02265554 l999-03- 16PCTlUS97I16718highly inhibitory to substitutions (83). In contrast,the first five nucleotides were relatively tolerant ofa number of substitutions and two of them at positionfour were up-regulatory mutations, resulting in a four-to 20-fold increase in RSV-CAT RNA replication andtranscription. Using a bi-cistronic minirepliconsystem, gene—start and gene-end motifs were shown to besignals for mRNA synthesis and appear to be selfcontained and largely independent of the nature ofadjoining sequence (84).The L gene start signal lies 68 nucleotidesupstream of the M2 gene-end signal, resulting in geneoverlap (Fig. 3) (74). The presence of the M2 gene-endsignal within the L gene results in a high frequency ofpremature termination of L gene transcripts. Fulllength L mRNA is much less abundant and is made whenthe polymerase fails to recognize the M2 gene-endmotif. This results in much lower transcription of LmRNA. The gene overlap seems incompatible with a modelof linear sequential transcription. It is not knownwhether the polymerase that exits the M2 gene jumpsbackward to the L gene—start signal or whether there isa second, internal promoter for L gene transcription(74). It is also possible that the L gene isaccessible by a small fraction of polymerases that failto start transcription at the M2 gene—start signal andslide down the M2 gene to the L gene—start signal.The relative abundance of each RSV mRNAdecreases with the distance of its gene from thepromoter, presumably due to polymerase fall-off duringsequential transcription (80). Gene overlap is asecond mechanism that reduces the synthesis of fulllength L mRNA. Also, certain mRNAs have features thatmight reduce the efficiency of translation. Theinitiation codon for SH mRNA is in a suboptimal KozakSUBSTITUTE SHEET (RULE 26)1?10152025W0 98/ 13501CA 02265554 l999-03- 16PCT/U S97/ 16718-45-sequence context, while the G ORF begins at the secondmethionyl codon in the mRNA.RSV RNA replication is thought (74) to followthe model proposed from studies with vesicularstomatitis virus and Sendai virus (16,81). Thisinvolves a switch from the stop-start mode of mRNAsynthesis to an antiterminator read-through mode. Thisresults in synthesis of positive sense replication-intermediate (RI) RNA that is an exact complementarycopy of genomic RNA. This serves in turn as thetemplate for the synthesis of progeny genomes. Themechanism involved in the switch to the antiterminatormode is proposed to involve cotranscriptionalencapsidation of the nascent RNA by N protein (16,81).RNA replication in RSV like other nonsegmentednegative—strand RNA viruses is dependent on ongoingprotein synthesis (85). Predicted RI RNA has beendetected for the standard virus as well as RSV-CATminigenome (74,85). RI RNA was 10-20 fold lessabundant intracellularly than was the progeny genomeboth for the standard and the minigenome system. Thenucleotide sequences (in positive strand, antigenomic,message sense) of various wild—type, vaccine andrevertant RSV strains, as well as the deduced aminoacid sequences of the RNA polymerase (L protein) ofthese RSV viruses, are set forth as follows withreference to the appropriate SEQ ID NOS. containedherein:SUBSTHUTESHEET(RULE26)?101520253035CA 02265554 l999-03- 16W0 98/13501 PCT/US97/16718_ 45 _Virus Nucleotide Sequence L Protein SequenceWild-Type2B SEQ ID NO:23 SEQ ID NO:2418537 SEQ ID NO:25 SEQ ID NO:26Vaccine2B33F SEQ ID NO:27 SEQ ID NO:282B20L SEQ ID NO:29 SEQ ID NO:30Revertant2B33F TS(+)2B2OL TS(+)SEQ ID NO:31SEQ ID NO:33SEQ ID NO:32SEQ ID NO:34Each RSV virus genome encodes an L proteinthat is 2,166 amino acids long. Genome length andother nucleotide information is as follows:Virus GenomeWild-Type Lenqth L Start Codon L Stop Codon2B 15218 8502-8504 15000-1500218537 15229 8509-8511 15007-15009Vaccine2B33F 15219 8503-8505 15001-150032B20L 15219 8503-8505 15001-15003Revertant2B33F TS(+) 15219 8503-8505 15001-150032B20L TS(+) 15219 8503-8505 15001-15003As detailed in Example 3and 8) below,subgroup B 3' genomic promoter region are nucleotide 4(C —>G), 5(especially Tables 7the key attenuating mutations for the RSVand the insertion of an additional A in thestretch of A's at nucleotides 6-11 (in antigenomicSUBSTITUTE SHEET (RULE 26)1?101520253035W0 98ll350lCA 02265554 l999-03- 16PCT/US97/ 16718message sense). As also detailed in Example 3 below,the key potentially attenuating sites for the L proteinof RSV are as follows: amino acid residues 353(arginine -9 lysine), 451 (lysine -9 arginine), 1229(aspartic acid -9 asparagine), 2029 (threonine -9isoleucine) and 2050 (asparagine —+ aspartic acid). Itis understood that the nucleotide changes responsiblefor these amino acid changes are not limited to thoseset forth in Example 3 below; all changes innucleotides which result in codons which are translatedinto these amino acids are within the scope of thisinvention.The attenuated viruses of this inventionexhibit a substantial reduction of virulence comparedto wild-type viruses which infect human and animalhosts. The extent of attenuation is such that symptomsof infection will not arise in most immunizedindividuals, but the virus will retain sufficientreplication competence to be infectious in and elicitthe desired immune response profile in the vaccines.The attenuated viruses of this invention maybe used to formulate a vaccine. To do so, theattenuated virus is adjusted to an appropriateconcentration and formulated with any suitable vaccineadjuvant, diluent or carrier. Physiologicallyacceptable media may be used as carriers. Theseinclude, but are not limited to: an appropriateisotonic medium, phosphate buffered saline and thelike. Suitable adjuvants include, but are not limitedto MPL" (3—O—deacylated monophosphoryl lipid A; RIBIImmunochem Research, Inc., Hamilton, MT) and IL-12(Genetics Institute, Cambridge, MA).In one embodiment of this invention, theformulation including the attenuated virus is intendedfor use as a vaccine. The attenuated virus may be mixedSUBSTITUTE SHEET (RULE 26)?101520253035W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/16718with cryoprotective additives or stabilizers such asproteins (e.g., albumin, gelatin), sugars (e.g.,sucrose, lactose, sorbitol), amino acids (e.g., sodiumglutamate), saline, or other protective agents. Thismixture is maintained in a liquid state, or is thendessicated or lyophilized for transport and storage andmixed with water immediately prior to administration.Formulations comprising the attenuatedviruses of this invention are useful to immunize ahuman or animal subject to induce protection againstinfection by the wild-type counterpart of theattenuated virus. Thus, this invention furtherprovides a method of immunizing a subject to induceprotection against infection by an RNA virus of theOrder Mononegavirales by administering to the subjectan effective immunizing amount of a vaccine formulationincorporating an attenuated version of that virus asdescribed hereinabove.A sufficient amount of the vaccine in anappropriate number of doses must be administered to thesubject to elicit an immune response. Persons skilledin the art will readily be able to determine suchamounts and dosages. Administration may be by anyconventional effective form, such as intranasally,parenterally, orally, or topically applied to anymucosal surface such as intranasal, oral, eye, vaginalor rectal surface, such as by an aerosol spray. Thepreferred means of administration is by intranasaladministration.In another embodiment of this invention, anisolated nucleic acid molecule having the completeviral nucleotide sequence of either the wild-typeviruses or vaccine viruses described herein is used togenerate oligonucleotide probes (from either positivestrand antigenomic message sense or negative strandSUBSTHUTESHEET(RULE26)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718complementary genomic sense) and to express peptides(from positive strand antigenomic message sense only),which are used to detect the presence of those wild-type virus and/or vaccine strains in samples of bodyfluids and tissues. The nucleotide sequences are usedto design highly specific and sensitive diagnostictests to detect the presence of the virus in a sample.Polymerase chain reaction (PCR) primers aresynthesized with sequences based on the viral wild-typeor vaccine sequences described herein. The test sampleis subjected to reverse transcription of RNA, followedby PCR amplification of selected cDNA regionscorresponding to the nucleotide sequence describedherein which have nucleotides which are distinct for adefined strain of virus. Amplified PCR products areidentified on gels and their specificity confirmed byhybridization with specific nucleotide probes.ELISA tests are used to detect the presenceof antigens of the wild-type or vaccine viral strains.Peptides are designed and selected to contain one ormore distinct residues based on the wild-type orvaccine sequences described herein. These peptides arethen coupled to a hapten (e.g., keyhole limpethemocyanin (KLH) and used to immunize animals (e.g.,rabbits) for the production of monospecific polyclonalantibody. A selection of these polyclonal antibodies,or a combination of polyclonal and monoclonalantibodies can then be used in a “capture ELISA" todetect antigens produced by those viruses.Samples of the Moraten measles virus vaccinestrain were deposited by Applicants with the AmericanType Culture Collection, 12301 Parklawn Drive,Rockville, Maryland 20852, U.S.A., under the provisionsof the Budapest Treaty for the Deposit ofMicroorganisms for the Purposes of Patent ProceduresSUBSTITUTE SHEET (RULE 26)?1015202530W0 98/ 13501CA 02265554 l999-03- 16PCT/US97/16718("Budapest Treaty") and have been assigned ATCCaccession number VR2587. Samples of the HPIV—3 virusVero-grown cp45 vaccine strain were deposited byApplicants with the American Type Culture Collection,12301 Parklawn Drive, Rockville, Maryland 20852,U.S.A., under the provisions of the Budapest Treaty andhave been assigned ATCC accession number VR2588.Samples of the 2B wild-type RSV virus were deposited byApplicants with the American Type Culture Collection,12301 Parklawn Drive, Rockville, Maryland 20852,U.S.A., under the provisions of the Budapest Treaty andhave been assigned ATCC accession number VR2586.Given these three deposited strains and thesequence information for these and other strainsprovided herein, one can use site-directed mutagenesisand rescue techniques described above to introducemutations (or restore a wild-type genotype) of all thestrains described herein, as well as taking thesestrains and making additional mutations from the panelof mutations set forth in Tables 3, 4 and 6-8 below.In order that this invention may be betterunderstood, the following examples are set forth. Theexamples are for the purpose of illustration only andare not to be construed as limiting the scope of theinvention.ExamplesStandard molecular biology techniques areutilized according to the protocols described inSambrook et al. (86).SUBSTHUTESHEET(RULE26)1?1015202530._..t.......................-..................‘..... .., .W0 98/13501CA 02265554 l999-03- 16PCT/U S97/ 16718Example 1MeaslesMoraten MV vaccine virus was grown once,directly from the Attenuvax” vaccine vial (Lot #0716B),the Schwarz vaccine virus was grown once (Lot96G04/M179 G4lD), while the Zagreb and Rubeovax”vaccine viruses were each grown twice in the Vero cellsbefore RNAs were made for sequence analysis. MVwildtype isolate Montefiore (56) was passed 5-6 timesin Vero cells before extraction of RNA materials andsimilarly, MV wildtype isolates 1977, 1983 (14) weregrown 5-7 times before extracting materials foranalysis. Edmonston wild-type isolate received fromDr. J. Beeler (CBER) (see Fig. 1) was the originalEdmonston isolate already passaged seven times in humankidney cells and three times in Vero cells beforereceipt and further passaged once in Vero cells beforeusing for sequence analysis.RNA was prepared by infecting Vero cells at amultiplicity of infection (m.o.i.) of 0.1 to 1.0 andallowed to reach maximum cytopathology before beingharvested. Total RNA from measles virus-infected cellswas extracted using Trizol” reagent (Gibco-BRL).The total RNA isolated from Vero cell passagematerial was amplified by the Reverse Transcriptase-PCR(Perkin-Elmer/Cetus) procedure using measles (EdmonstonB strain (19)) specific primer pairs spanning the 3'and 5' promoter regions and the L gene of the viralgenome. Table 2 presents these primer sequences. Theprimers of SEQ ID NOS:35—54, 74, 77 and 78 are inantigenomic message sense. The primers of SEQ IDNOS:55-73, 75, 76 and 79 are in genomic negative-sense.SUBSTITUTE SHEET (RULE 26)_.. ...N..._............-—-.........-..... . . ......_.H... ‘I51!-<> V?CA 02265554 l999-03- 16W0 98/ 13501 PCT /US97/16718Table 2Primers for PCR and Sequencing MV L Genesand Genomic TerminiMuCATATCACTCACTCTGGGATGGAGWW (SEQ ID NO:35)”nTCAGAACATCAAGCACCGCC”n (SEQ ID NO:36)”“ACAGTCAAGACTGAGATGAGnw (SEQ ID NO:37)1WMAAGAGTCAGATACATGTGGANMO (SEQ ID NO:38)1“QACATGAATCAGCCTAAAGTCMNO (SEQ ID NO:39)1“HCCGAAAGAGTTCCTGCGTTACGACCmwe (SEQ ID NO:40)IN“CAGTCCACACAAGTACCAGGHNZ (SEQ ID NO:41)1“MGTCAGAAGCTGTGGACCATCHH0 (SEQ ID NO:42)1m“AATATTGCTACAACAATGGCnun (SEQ ID N0:43)in%ACTCTTCATTCCTAGACTGGgns (SEQ ID NO:44)1%”GTCCAATTATGACTATGAACH“1 (SEQ ID NO:45)12891AGAACAGACATGAAGCTTGC1291o (SEQ ID NO:46)1”nCCAACAAGGAATGCTTCTAGu?1 (SEQ ID NO:47)1”“ACAGCACTATCTATGATTGACCTGGH95 (SEQ ID NO:48)1”wGCAACATGGTTTACACATGCuus (SEQ ID NO:49)1”WAGATTGAGAGTTGATCCAGGun9 (SEQ ID NO:50)1“”AGGAGATACTTAAACTAAGCu“R (SEQ ID NO:51)1”uTAAGCTTATGCCTTTCAGCG?wo (SEQ ID NO:52)1533.,TTAACGGACCTAAGCTGTGC15356 (SEQ ID NO:53)1“HGAAACAGATTATTATGACGGHno (SEQ ID NO:54)nWCGGGCTATCTAGGTGAACTTCAGG”H (SEQ ID NO:55)”WATTTGGATATGGAATATGAG“u (SEQ ID NO:56)”wACTCAACTGAACTACCAGTGnu (SEQ ID NO:57)InMAAGAACATCATGTATTTCAGmu2 (SEQ ID NO:58)1WHTTATCAACGCACTGCTCATGNB0 (SEQ ID NO:59)1”HATTTTCAGCAATCACTTGGCATGCCmm5 (SEQ ID NO:60)1””GCCTCTGTGCAAACAAGCTGn%1 (SEQ ID NO:61)1””TCTCTAGTTACTCTAGCAGCHH, (SEQ ID NO:62)inMAGGTCGTTGTTTGTGAGGAGHH1 (SEQ ID NO:63)H3HTCGTCCTCTTCTTTACTGTCHMZ (SEQ ID NO:64)SUBST?UTESHEET(RULE26)I?1015202530W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718_ 53 _1“”CCGTCCTCGAGCTAGCCTCGumo (SEQ ID NO:65)1”HCTCCTCCAGGCTCACATTGGBW3 (SEQ ID NO:66)1MMGGGTTGGTACATAGCTCTGCHW1 (sEQ ID NO:67)1”MCACCCATCTGATATTTCCCTGATGGBH3 (SEQ ID NO:68)1“MTGGTTGACAGTACAAATCTGHMo (SEQ ID NO:69)1“WCTGAAATGGGAAGATTGTGCHH1 (SEQ ID NO:70)M8,OAGCAATCTACACTGCCTACCM801 (SEQ ID NO:7l)lawTCACAGATGATTCAATTATCuu1 (SEQ ID NO:72)155,OGATCCTAGATATAAGTTCTC15511 (SEQ ID NO:73)1ACCAAACAAAGTTGGGTAAGG21GGGGGATCC1,,OATCCCTAATCCTGCTCTTGTCCCN,20DGATTCCTCTGATGGCTCCACH1 (SEQ ID No:76)15,,1TAAcAG'1'cAAGGAGAccAAAG,_5,,,, (SEQ ID NO:77)GGGAAGCTT15”1AACCCTAATCCTGCCCTAGGTGG15323 (SEQ ID NO:78)15,,,4ACCAGACAAAGCTGGGAATAGP-15373 (SEQ ID NO:79)(SEQ ID NO:74)(SEQ ID NO:75)Overlapping PCR fragments of the completeviral genome were directly sequenced without cloning toachieve the consensus sequence, by the dideoxyterminator cycle sequencing method using both strands(ABI PRISM 377 sequencer and ABI PRISM sequencing Kit).To determine the sequence at the absolute termini, aligation procedure described previously was used (55).To test this hypothesis, the nucleotidesequences were determined for the non—protein codingregulatory regions and the L gene of the progenitorEdmonston wild—type MV isolate, for the availablevaccine strains derived from this isolate, as well asfor other wild—type strains. Nucleotide (inantigenomic, message sense) and amino acid differenceswere then compared and aligned as set forth in Tables3-5 (differences are in italics):SUBSTITUTE SHEET (RULE 26)_ . ..._.............—-——..........(.?CA 02265554 l999-03- 16W0 98/ 13501 PCT/US97/ 16718Table 3Differences in MV 3' Genomic Promoter RegionNucleotide SequenceNucleotide number:Virus gg gg gg ggEdmonston w-t A A G GVaccines:Rubeovax” T C G GMoraten T C G GSchwarz T C G GZagreb T T G AAIK-C T C G GWild-Types:1977' A A A G1983 A A A GMontefiore A A A GSUBSTHUTESHEET(RULE26)I?W0 98/13501CA 02265554 l999-03- 16PCT/US97/16718Table 4Differences in MV L Nucleotides and Amino AcidsBetween Edmonston Wild-Type and Vaccine StrainsEdmonston w—tMutationEdmonston w-tRubeovax” vac.Moraten vac.Schwarz vac.Zagreb vac.AIK—C vac.0)DJI-'uh0lo1624 1649 1717 1887 1936 2074 2114ATT GCA ACC AGG GAT AAC CAT CAA AGAACT ACA GCC ATG GCT GAC TAT CGA AAAHH|~]|~]HH'~3'~J3='{vtl='.'>'as-3»-3+5»-1»;wwzzzw> b knit > Uzzuutvz~<:=:::::::::::r:auaoaozoaoosuwrqwwwSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16PCT/US97/16718W0 98/1350]-56..H n 2 > 2 > 22 n n > 2 H 22 2 n H o H 22 n m > o H 2<94 H22 242 «am 220 «aw «mme92 H4w umw «Hm 24o «H2 mooomm m¢m Hwm Hmm mmm «mm «mm2 2 2 n 2 2 H2 2 2. 2 2 2 H2 2 2 n 2 2 >2 2 > n 2 2 Ho2u 222. mew H4< «a2 «a2 HBUuma 222 new 92m 222 224 994mHm ohm mHm com H¢¢ H«« HmnE-‘E-IE-Id}¢UU?vdN m Nmnwmnum 0mhH-UHw3 nmw3uwmmmuaod 02.124 ?aw mmmuwuomaodz _H E ._H..n mmunmnwmmwnm mHnmH.u-3 mnowmmunosu-3 mmmHuu3 2.3u-3 noumno?vmGOHUMUUSU I3 HHOU MGOEUHps3 mnowmounozuu3 $3u-3 pumaps3 noumno?vmnoaumunzU I3 AHOU WHHOFHUWSUBSTITUTE SHEET (RULE 25)?CA 02265554 l999-03- 16PCT /U S97/ 1 6718W0 98/1350]_57_> #1 >M!-KNH P1 >Z 3: mEll-IHH> MH2U?d 000 ¢aw BUd Hhd H00 U60 Bd? UdH d??QCDQDQDLDQE >4 NMintuit:mH>>>u-3 onoammunozu-3 mmmHu-3 bbmHu-3 noumno?vmGOMUNUUSUBO Odd dBd Edd Bud Bdw Udw emu UHF dHU u-3 noum?o???mmam mHHN mmom omen saga MHON boom mmma mmma ommaZ Z InH rd >> *4 >U)!-umU (9 Md to d2 H > m 0 dH0d UHU B?d. UUd ¢mw duhHdd U?d BBO Uwd «aw UUUmwua mona mmma vmNH wwoa ohmM MM M0 RM NUdU UUHU00 U00¢Hm wanmn?muum mmhB-UHw3 Gmm3ummmµ.nUd O.—H._...?H¢. wnm MOUHUOUHUUZ ..H >2 HHH mwunwuwmmunn.uonn?»uoov m magmau-3 UHOMHGUGOSu-3 mamapa? pumau-3 noumdo?um?OHUMu5Su-3 noumno?vmSUBSTHUTESHEET(RULE26)?CA 02265554 l999-03- 16W0 98/ 13501 PCT/US97/16718Example 2PIV-3A comparison of sequences (in antigenomic5 message sense) of the parental wild—type JS strain ofPIV-3 virus and the FRhL-grown and Vero-grown forms ofthe cp45 mutant are set forth in Table 6. Where acodon change does not result in an amino acid change,Table 6 states “none”, followed by the name of the10 unchanged amino acid.SUBSTITUTE SHEET (RULE 26)I’?_1_l_;CA 02265554 1999—03-1(,l~3‘_‘H 7'.J‘.m‘.m—- -rt! MU ‘_':m:»I»H..‘).;/I-.-.A.H77:1.“-W'Il’.\-\l| §'j\ClI|:\4 .wv. x.w.IRL‘\. \0\ II?§0H.U?U5i. micm onEmma. can T n5. .54 T .54 u. H U mannaAuamnu 0?m\0§O? FEB al UB9 9 B U aamna.23 0.3 T 504 9.5. T 0.5. H. H. G Hub:3%. 3: T man. 040 T 92. u u 9 3+5aN##. 5HU\u:O: U46 4! 440 0 U 4 dbmmGag .H~wH.\0.FOC U5... +1 Eda. U U H. mnmm unwvoo A6.: T Hub. HUG +1 Ham U U H uni.hmwxvcon UUU T HUG U U H. 2.3 mamvoo .2!use T HA4 404 T «U0 4 4 U mH¢wHu> T OHH 490 T 65 U 0 d mwmwEixonon .52 T 32 H H u emmm m??oo N Huse T 85 00¢ T uuu 4 4 u 5.3. meanou 2 WW.dudxo?on U44 41 Hand U U H coon mdwvou A D«Q T uom sou T BUB u u H 3.3 cmmad T dab U00 7 UBO U U H van mcmdou AZ W.B H 4 3 Fr: .5 W.1 4 B 3H B 0 anF B U ?nU N U a. mu uanuua .m3 cm M03355 mvnu mvau ?owuqnom .0m:mAU UMU4 O?w?? om?usu no?mmva oHo> a?mh ma ??muoodu?z doumo? 0:00ndmmuuu ma 4 mvmo .:.3OHmI..~..—~wh was uouub no :OnmH.n9nOU ou?o?vomw UHQMH.?V..-RC\. \<)\-|'.l’\-\H l‘.\Ll”‘..\-w -4‘-". .-J‘Jl.‘.'- so -sequence analysis of the parental wild-typeJS strain oi PIV-3 virus and the FRhL-grown cp45 mutantshowed that the latter contained 20 nucleotide changes.Four changes were in the noncoding 3'-leader region at'5 nucleotide positions 23 (T -ic). 24 (C -91). 28 (G -+T) and 45 (T ~+ A) (in antigenomic, message sense).when considered in the genomic, negative sense, thechange at position 28 from the smaller pyrimidine (“C”)to the larger purine (“A”) may change the size of the10 region tlanked by the conserved regions of the 3'genomic promoter region. resulting in an alteredspatial presentation of the cis-acting signals to thepolymerase.Nine changes were coding changes in the NP.15 M, F, HN and L genes. The other seven changes werenon-coding or silent changes in the NP, 9, F, KN and Lgenes or the NP untranslated region (UTR). The cp45mutant has been demonstrated to have poor transcriptionactivity at non-permissive temperatures due to its ts20 phenotype (87). This ts phenotype has now been mappedto the viral L gene (88). Because the cp45 virus hasbeen shown to function normally with regard tomutations in the HN and F glycoproteins (87), thissupports the implication that mutations in the 3'-25 leader and L gene contributed to the attenuatingphenotype of this virus.Thus, the four 3' leader specific changes inFRhL-grown cp45 and the three coding changes in the Lgene at amino acid positions 942 (Ty: -+His), 992 (Leu30 ->Phe) and 1558 (Th: -+Ile) contributed signiticsntlyto the attenuation phenotype of the candidate cp45vaccine strain.AMENDED SHEETw? =27-llCA 02265554 1999-03-lowxcu TnHunM~ r1H Nu ;»uu+4n: 7 r‘_ ‘ .l..-l._.Aj“ _‘-_‘_\. _ ' "’ '?ucy\o\wun-mImcm9.nS'_ .I‘.'.1..\J‘.a A.-.‘\-101520’ 127-ll-(LIA 02265554 1999-03-161"-‘~'H :'U‘.H.".n<1«\.....A. 51 -The first two amino acid changes in the Lprotein (at positions 942 and 992) map to one of thehighly conserved arses among all Paramyxovirus L genes.The third amino acid change (at position 1558) maps tothe area joining two conserved blocks corresponding tothe change at anlnc acid 1717 in the 3? Vaccinestrains.The published literature (89) sets forth only18 changes between the antigenomic message sensesequences ot the JS and Fnhn-grown cp45 strains.Sixteen of these changes were found by applicants.The published literature did not report fourchanges found by applicants: in the 3' leader atnucleotide 45 (T -+ A), in the NP UTE at nucleotide 62(A -9 T), or the changes in amino acids in the NPprotein resulting from the changes at nucleotide 397 (T-+ C), leading to the amino acid change (Val -oala) andnucleotide 1275 (T —aG), leading to the amino acidchange (Ser ~+Ala) (nucleotide changes in antigenonic.message sense).AMENDED SHEETg Hz) 25:) 2:11:29-l--1n.‘;:n I?1015202530WO 98113501ICA 02265554 l999-03- 16PCTIUS97/16718Example 3RSV Subgroup BThe temperature-sensitive (ts) phenotype isstrongly associated with attenuation in vivo; inaddition, some non-ts mutations may also beattenuating. Identification of ts and non-tsattenuating mutations was achieved by sequence analysisand evaluation of ts, cold-adapted (ca), and in vivogrowth phenotypes of RSV mutants and revertants.The genomes of the following five RSV 2Bstrains have now been completely sequenced: 2B parent,2B33F, one revertant designated 2B33F TS(+), 2B20L andone revertant designated 2B20L TS(+). The 2B33F and2B2OL strains are ts and ca and are described in U.S.Serial No. 08/059,444 (90), which is herebyincorporated by reference. After identifying regionswhere mutations in 2B33F and 2B20L are located, nineadditional isolates of 2B33F "revertants" obtainedfollowing in vitro passaging at 39°C and in vivopassaging in African Green Monkeys or chimpanzees, andnine additional isolates of 2B20L "revertants" obtainedfollowing in vitro passaging at 39°C have beensequenced in those regions. The ts, ca, andattenuation phenotypes of many of these revertants havenow been characterized and assessed. Correlationsbetween phenotype ts, vaccine attenuation and sequencechanges have been identified.A sumary of results is presented in Tables7-12.SUBST?UTESHEET(RULE26)?W0 98/13501CA 02265554 l999-03- 16-63-Table 7PCT/US97/16718Sequence comparison between RSV 2B and 2B33F strainsFor 2B33F and 2B33F TS(+), nucl.are one larger than for 2B for M, SH & L genesNucl. Nucleotide changespos.fGene/ 3‘ end RSV 2B RSV’ RSV 2B33F Amino acidregion of VRNA 2B33F TS(+), 5a changesrevertantGenomic 4 C G G non - codingPr“““°r 6 - extra A extra A non—codingM 4175 T C C non—coding4199 T C C non—codingSH 4329 '1' c c Phe-Leu (10)4409 T C C none Ile (36)4420 '1' c c Ile-Thr (40)4442 T C C none His (47)4454 T ¢ C none Cys (51)4484 T C C none Tyr (61)4497 T c c Stop—Gln (66)4505 T C C none Ser (68)4525 T C C Ile-Thr (75)4526 T C C Ile-Thr (75)4542 T C C Stop-Gln (81)4561 '1‘ c c Leu-Pro (87)4575 T C C Trp-Arg (92)4598 T C C none Thr (99)L 9559 G A A Arg—Lys (353)9853* A G A Lys—Arg (451)*12186 G A A Asp-Asn (1229)14587 C '1' '1' Thr—Ile (2029)15071 A G G non—codingpos. numbersAt pos. 9853, the Lys—Arg change has revertedback to Lys in the 2B33F TS(+) strain \SUBSHTUTESHEET(RULE25)?CA 02265554 l999-03- 16W0 98/13501 PCT/US97/16718Table 8Sequence comparison between RSV 2B and 2B20L strainsNucl. Nucleotide changespos.fGene/ 3' end RSV 2B RSV RSV 2B2OL Amino acidregion of VRNA 2B2OL TS(+), R1 changesrevertantGenomic 4 C G G non — coding*P"°“‘°t°r 6 - extra A extra A non-coding*L 8963 C T T none Thr (154)13347 A A G Asn-Asp (1616)14587 C T T Thr—I1e(2029)*14649 A G G Asn-Asp (2050)14650 A A T Asn-Asp-Val(2050)**1' For 2B20L and 2B2OL TS(+), nucl. pos. numbersare one larger than for 2B for L gene* Mutation is common in 2B33F and 2B20L strains** At pos. 14650, the mutation suppresses the tsphenotype in 2B20L TS(+) revertantSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16PCT/US97/16718W0 98/ 13501_ 55 _Uonm um umxuwn onumam;\$ $\.C 35.1%. nun.» Human >U.N¢#Dz Dz nw.H no.NW «$8.0 m.o 26¢ vwuomu?wnhmmmn Nmm SE4.w\v. .¢\m. .az. uomm um umxu?m msumam A+.maDz Oz .m.~ .m.uv S... m.o Jmmmmmm nmncwmm hmmmm an - hmmmm >mm3.}; 3&3 3.5 Uomm um vmxu?m wzumam Alma.Dz Dz .m.m L. .nv 36 n.o Jmmummm Hwnc?mm mmmmm mv n mmmmn >mm:13 213 3.\$ 333 3.2: Uomm um @933 wsvmam Time.uo.¢ .~.w .m.m >.H| . . .. v me o m o wmmmwmm umcnwmm mmmmm mm hnmmm bmm.w\o. Aw\H. -» :mwxcv .«.v. N.H m.Hv an a. \. n . v .. . I .u3\uc.m\mmv uwmmmmmmuuaoo mm Scum.m ov ¢o m .m av .m av ¢o.o >oooo.owuumaonun uumua? mu ‘mu hmmmm >mM.¢\«. .«\¢v nN.m nm.M .nn..HM..HumL..v .m.m .w.m .m.m Soc... ~..o unwumm mmhunvanz. mm >mmwmm>ma sum: mwumuanuau uam?r nmuoa wsvmamHmunnonoum Hmwmz mmnda Hmmmz Uommxom mow UoNm\mmS64 umm couuou mu muEusouu o>..n> GH omhuodmnm oNu..n> «NH muhsom wam?mmmnwmnum uGmuHo>mm vnm mu .mmm manna>mmSUBS?TUTESHEET(RULE26)?CA 02265554 1999-03-16PCT/US97/ 16718WO 98/13501_ 55 _Awxv. .w\m. Aazv uonm um uwxo?m osvmam. . | . nmzu Hana: can .mmmH¢nz nz uo m an mv oz » o ma?ao uouumunH-mmmmu «mum ma?nuDonn um voxu?m msvmam.v\m. .«\«. .az. omm>mH: Hmmnomuu mu .ommHu3- n noz oz 0 mv u¢ N oz 5 o ma?nu wouomu:?-mmmm~ «mum ms?nouonm um vwxo?m osmmam0mm>manz oz nz nz nz .a¢V Hmonumuu vo .~mmH# mm m am o mawnu oouowm:?-mmmm~ «H e.nuUomm um vmxuwm msvmam.¢\o. .¢\H. .uz\u:?\mm. awn; Ham»: aHv.w<unz nz no.Hv nv.HW noo.o vooooo.o sea uwuomw:«-mmmm~ ham zwmcomm um umxo?m wswm?m.«\o. .«\HV .u:V nmma Hanna ~Hu.«¢anz oz aH.Hv nm.HW moooo.ow ¢ooooo.o 294 uouumuc«-mmmm~ man 204Uoum um Uwxounm mdwmam.v\¢. .w\o. .u:?.mm. nuns Hana: >u.~¢*nz az n~.H nw.Hv moo.o H.o :w4 uouowu:w-mmmm~ «mm sum0mm>m.H nmmz mmumnannsu uawww nmuo? wnvmdmHm?nucoum Hmmmz manna Hmmmz oonmxom mom uo~m\mm=w« umx nouuoo an an«nuzouw o>.._...> 2H mmhuonwnm o.nu.....> GH wounom mam?mmmnwmnum uamuHm>mm was mu .mNAumsa?uaouv m magma>mmSUBSTITUTE SHEET (RULE 26)?CA 02265554 £999-03-16PCT/US97ll6718WO 98/13501wnwm oaumam ?u?????h?u?? n new- 67 _HH+zH mum cgoa nZH ohm neon uwmon «mmon umuwm wsvmam Hanan nAamuou #\vwuowwa« my umuwu msuw> nmo?mmHH+ZH bum mgoaza bum .aoHwnwm msvmam mm>u-UHH3 n asmwon owmon a2n bum rmoazu amm haoawmon ummon .0:00 no: u DzSmoa cw Uwusmmo? £u3oum o>M> aH «can an «.9333&3 3&3 E: mzu?m tmmmmmmm Emanz oz uN.m um.m oz ».o nwncwmu gamma cam Aommm >mmcomm um E333Av\vV Avxmv .HzV wnvmam .mmmmmmm A+VmHnz nz uo.v ou.~w oz m.o uwnc?mm gamma mm gamma >mmcan an umxuwmAv\wv Av\mV Ahzv mnvmam .wmmmmmm A+vmHnz nz yuan um.~w nz m.o ummmwmm new N mm qommu >mm9.3 an uwxu?mAv\v. Av\vv ABZV wsbm?m .mmmmmmm A+.mHnz nz um.m um.~ oz m.o uwnu?mm momma Hm aommn >mxon X uwmmmmmm-uHooG): S): 3.): 3}: Bs\uu? mm scum uoumao?nu>.ov u>.ov um.av om.Hv mo.o Nooo.o ucuusa mu .mu Aommm >mmomm>nA nmmz mmumcwnnsu GHOMN nmuo? wnvmamH?M£UEOHm Hmmmz mmasq Hmmmz UoNm\oN mom oommxmm20¢ umm nouuou mu muasuxonu o>w> nH wmxuonmnm ouuw> EH munuom ?????mmcwmnum unmunmbwm VCM WuGmsn?nouv m manna.mm >mmSUBSTHUTESHEET(RULE26)................._.................,A...... , ,?CA 02265554 l999-03- 16W0 98/ 13501 PCT/US97/ 16718Table 102B33F Revertantsts (+) In vitro AGM Chimp5a 4a 3b pp2 pp4 pp6 pp7 1A 3A 5Abase no.T___AL___4176,4200 S S S S S S S S S SSH14 bases* S S S S S S S S S SL9560 S S S S S S S S S S9854 2B 2B 2B 2B S S S ND 2B 2B12187 S S S S S S S S S S14588 S S S S S S S ND S S15072 S S S S S S S S S SPhenotypets 2B 2B 2B r r S S 2B 2B 2Bca S S S 2B S 2B S ND ND NDAttenuated r r r (r) (r) S S r r1 These 2B33F revertant base nos. are one larger than for 2B for M,SH and L genes* bases 4330,4410,4421,4443,4455,4485,4498,4506,4S26,4527,4543.4562,4576,4599S = same base as 2B33F2B = reversion to 2B base or complete reversion in phenotyper = moderate reversion in phenotype(r) = slight reversion in phenotypeND = not doneSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/ 13501 PCTIUS97/16718-69..Table 112B2OL RevertantsTS(+) In vitro Isolatesbase no , 1' R1 R2 R39. R4A RSA R6A R715. R8A R9A RIOA__JL__8964 S S S S S S S S S S13348 C* S ND S S ND S S S S14588 S S S S S S S S S S14650 S S 2B S 2B 2B S S 2B 2314651 A* A* S A* S S A* A* S SPhenotype(:8 2B 2B ND ND ND ND ND ND 2B 2BAttenuated r r ND ND ND ND ND ND r r1 These 2B20L revertant base nos. are one larger than for 23 for LgenesS = same base as 2B2OL2B = reversion to 2B baser = moderate reversion in phenotype* = base change, different from 2B or 2B2OLND = not doneSUBSTHUTESHEET(RULE26)?CA 02265554 1999-03-16WO 98/13501 PCT/U S97/ 16718_ 70 _Table 12RSV 2B, ts and Revertant Strains: Phenotype SummaryVirus Isolate Source In Vitro In VivoPhenotype Attenuationts ca Cotton AGMRatRSV 23 Wild-type Parent Strain - - — -RSV 2B33F ca, ts mutant isolated ++++ ++ ++++ +++from 2B, cold-passagedx 33RSV 2B33F - 5a 2B33F spinner passage — ++ ++ +TS(+) plaque picked at 39°CRSV 2B33F ' 43 2B33F spinner passage - ++ ++ NDTS(+) plaque picked at 39°CRSV 2B33F - 3b 2B33F spinner passage - ++ ++ NDTs(+) plaque picked at 39°CAGM pp2 2B33F-infected AGM A2, + - +++ NDd7 nasal wash plaquepicked at 32°CAGM pp4 2B33F—infected AGM A2, + ++ +++ NDd7 nasal wash plaquepicked at 32°CAGM pp6 2B33F-infected AGM A4, ++++ - ++++ NDdl2 nasal wash plaquepicked at 32°CAGM pp7 2B33F-infected AGM A4, ++++ ++ ++++ NDdl2 nasal wash plaquepicked at 32°Cchimp Pp1A 2B33F-infected chimp _ ND ND ND#1552, d4 tracheallavage, plaque pickedat 32°CChimp pP3A 2B33F-infected chimp _ ND ++ ND#1560, d6 tracheallavage, plaque pickedat 32°Cchimp PPSA 2B33F-infected chimp _ ND ++ ND#1563, dl0 tracheallavage, plaque pickedat 32°CSUBSTITUTE SHEET (RULE 26)1?CA02265554 1999-03-16W0 98I1350l PCT/US97/16718_ 71 _Table 12 (continued)RSV 2B, ts and Revertant Strains: Phenotype SummaryVirus Isolate Source In Vitro In VivoPhenotype Attenuationts ca Cotton AGMRatRSV 2B20L ca, ts mutant isolated ++++ ++ ++++ ++++from 2B, cold-passagedx 20RSV 2B2OL R1 2B20L spinner passage — ND ++ NDTS(+) plaque picked at 39°CRSV 2B20L R2 2B20L spinner passage - ND ++ NDTS(+) plaque picked at 39°CRSV 2B20L R9 2B20L spinner passage — ND ++ NDTS(+) plaque picked at 39°CRSV 2B20L R10 2B20L spinner passage - ND ++ NDTS(+) plaque picked at 39°CND = not doneadapted, not attenuated+ to ++++ =adaptation or attenuationSUBSTITUTE SHEET (RULE 26)_ wild-type phenotype, i.e., not temperature sensitive, not coldincreasing levels of temperature sensitivity, cold-?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/U S97/ 16718Several significant observations can be drawnfrom these data:a. As shown in Tables 7 (for 2B33F) and 8 (for2B2OL), there are relatively few sequence changesidentified in the two mutant strains: RSV 2B33Fdiffers from parental RSV 2B by two changes at the 3'genomic promoter region, two changes at the non-coding5'-end of the M gene, and four coding changes plus onenon-coding (poly(A) motif) change in the RNA dependentRNA polymerase coding L gene. In addition, 14 changesmapped to the SH gene alone. RSV 2B20L differs fromits RSV 2B parent only at seven nucleotide positions,of which three are common with 2B33F virus, includingtwo changes at the 3' genomic promoter and one codingchange in the L gene. Two additional unique changes of2B20L virus mapped to the coding region of the L gene.Potentially attenuating mutations at the non-coding 3'genomic promoter region and the RNA dependent RNApolymerase gene have been identified.b. Two ts mutations can be identified in the Lgene of the attenuated virus strains 2B33F and 2B20L:(i) In 2B33F, a mutation at nucleotide position9853 (A —>G) leading to a coding change in L proteinat amino acid 451 (Lys ->Arg) is clearly associatedwith the ts and attenuation phenotypes. Reversion atthis site alone in the 2B33F TS(+) 5a strain isresponsible for complete restoration of growth at 39°C(Table 9) and partial reversion in attenuation inanimals. This association with the ts and attenuationphenotypes was also supported by partial sequenceanalyses of six additional "full TS revertants"(designated 4a, 3b, pp2, 3A, 5a, SA) isolated from cellSUBS?TUTESHEET(RULE26)?1015202530.......-....;..........................r... wW0 98/13501CA 02265554 l999-03- 16PCT/US97/16718culture and from chimps, in which only the nucleotide9853 mutation reverted (Tables 10-12) (note that oneAGM (African Green Monkey) isolate which reverted at9853 only partially reverted in ts phenotype). Thisamino acid 451 mutation (Lys —>Arg) is amenable tostabilization in CDNA infectious clone constructs, byinserting a second mutation to stabilize the codon,thereby lessening the likelihood that it will revertback to Lys.(ii) In 2B20L, a mutation at base 14,649 (A —>G)leading to a coding change in the L protein (amino acidposition 2,050, Asn —>Asp) appears to be associatedwith the ts and attenuation phenotypes. This asparticacid at the amino acid 2050 invariably reverts back(Asp —>Asn) in TS(+) revertants or changes to adifferent amino acid (Asp —+ Val) by nucleotidesubstitution at position 14,650 (A —>T) (Tables 8,11). The above observation is based on completesequence analysis on the TS(+) revertant R1 and partialsequence of several additional TS(+) revertants (R2,R4A, R7A, R8A) at selected regions (Table 11). Anadditional mutation is seen in the R1 revertant atnucleotide postion 13,347 (amino acid 1616, Asn -9Asp) associated with the above reversion. However, theeffect of this mutation on the ts phenotype is notknown; the L gene of other revertants has not beensequenced completely.c. Three base changes are common to 2B33F and2B20L strains of virus:(i) A change at position 14,587 (C —>T) with acorresponding change (Thr —>Ile) at amino acid 2029 isSUBSTHUTESHEET(RULE25)?101520253035W0 98/13501CA 02265554 l999-03- 16PCT/US97ll6718present in both 2B33F and 2B20L (Tables 7,8). Thisnucleotide "T" substitution was found to be present in10% of the population of the progenitor RSV2B strainand may have been preferred during the attenuationprocess. No wildtype base "C" was found in the 2B33Fand 2B20L virus.(ii) Two mutations are seen in the 2B33F and 2B20L3' genomic promoter region: nucleotide 4 (C —>G) andthe insertion of an extra A in the stretch of A's atpositions 6-11 (in antigenomic, message sense). Whenthe sequences of selected TS(+) revertants wereanalyzed, these mutations were seen to have beenretained in the 2B33F TS(+)5a (Table 7) and the 2B20LTS(+)R1 (Table 8) revertants. These non—coding, cis-acting mutations remained associated with partial viralattenuation.Expression using the minireplicon RSV-CATsystem for the analysis of these cis—acting changes hasshown the 3' genomic promoter nucleotide 4 (C —>G)change to be an upregulation oftranscription/replication in this in vitro system whenthe 2B progenitor virus or either of the 2B33F or 2B33FTS(+) provided helper L gene functions (the N, P and M2genes are identical in these viruses).Complementation analysis of the 2B33F 3'genomic promoter and the helper functions provided bythe progenitor RSV2B virus or the 2B33F and 2B33F TS(+)viruses by this RSV-CAT minireplicon system has alsobeen conducted. All three viruses supported both the2B and 2B33F 3' genomic promoter mediatedtranscription/replication functions. However, the2B33F and 2B33F TS(+) viruses preferred their 2B33F 3'genomic promoters. This analysis clearly shows co-evolution of 3' genomic promoter changes during theSUBSTITUTE SHEET (RULE 26)I?101520253035WO 98113501CA 02265554 l999-03- 16PCTIUS97/16718- 75 -vaccine attenuation process, along with the RNAdependent RNA polymerase gene. Reversion of tsphenotype in the 2B33F mutant 5a by reversion of thesingle L protein amino acid 451 (Arg —>Lys) bysequence analysis was clearly demonstrated by supportof transcription/replication functions of RSV-CATminireplicon at 37°C. The 2B33F virus did not providehelper functions to the RSV-CAT minireplicon (with 2Bor 2B33F 3' genomic promoters) at 37°C.d. A biased hypermutation of SH seen in 2B33F ispresent in all 2B33F revertants, regardless ofphenotype, and is not seen in 2B20L, which is ts, ca,and attenuated. Thus, there are no data at this timethat associate this mutation with any biologicalphenotype.H Another wild—type RSV designated 18537 wasalso sequenced and compared to the sequence of thewild—type RSV 2B strain. With one exception, at allthe critical residues described above, the two wild-type strains were identical. For 2B, the codon ACA atnucleotides 14586-14588 encodes a Thr at amino acid2029 of the L protein, while for 18537, the codon ATTat nucleotides 14593-14595 encodes an Ile at amino acid2029 (the L gene start codon is at nucleotides 8509-8511 in 18537, compared to 8502-8504 in 2B).Example 4PCR Assay to Detect Measles Virus A 21 year old patient was admitted to ahospital with a three week history of progressive non-productive cough, shortness of breath, and fever. Hissymptoms failed to improve following treatment withclarithromycin for seven days or after a similar courseSUBSTITUTE SHEET (RULE 26)?101520253035WO 98/13501CA 02265554 l999-03- 16PCT/US97/ 16718of treatment with atovaquone. Concomitant complaintsof right upper quadrant abdominal pain provedrecalciltrant to omeprazole and antacids. Relevantpast medical history included Factor VIII deficiencyand HIV infection diagnosed 3-4 years prior to thishospital admission. One year earlier, he had receiveda booster immunization of measles—mumps-rubella (MMR)vaccine as required for college enrollment.Bronchoalveolar lavage and transbronchialbiopsies performed two days after admission to thehospital demonstrated reactive hyperplasia and alveolarlining cell desquamation with minimal chronicinflammation. No microorganisms were revealed by Gram,methenamine silver, or PAS stains. CT scans of thechest showed multiple, ill-defined, confluent nodulesat the left lung base. Despite administration ofempiric antimicrobials for opportunistic bacterial,mycobacterial, and fungal pathogens commonlyresponsible for pulmonary complications of advanced HIVdisease, the patient became and remained febrile to39°C. A 1eft—sided pleural effusion developed;diagnostic thoracentesis showed it to be exudative butotherwise non—diagnostic. Bronchoalveolar lavageperformed three weeks later only demonstrated alveolarhistiocytes, some of which were hemosiderin laden, afew lymphocytes, and neutrophils. FITE, AFB, andmethanamine silver stains again were negative.Two weeks thereafter, a wedge resection ofthe left lung was performed through CT~guidedminithoracotomy. Multiple tissue sections revealednodular areas of acute and chronic inflammation withregions of necrosis and fibrosis. Numerousmultinuclated giant cells were present, some of whichcontained both intracytoplasmic and intranuclearinclusions suggestive of measles virus giant cellSUBSTITUTE SHEET (RULE 26)I?CA 02265554 l999-03- 16W0 98/13501 PCT/US97/ 16718_ 77 _pneumonia. Special stains for bacteria, fungi, P.carinii, and acid fast organisms again gave negativeresults. Electron microscopic examination of sectionsof this lung biopsy revealed particles morphologically5 consistent with paramyxoviruses such as measles virus.Serum anti-measles IgM titers determined by a solidphase hemadsorbant assay were negative, as was asubsequent IgM capture immunoassay.Two weeks later, Rhesus monkey kidney (RMK)10 tissue culture cells inoculated with the patient's lungbiopsy material revealed cytopathic changescharacteristic of measles virus infection.Confirmation was obtained using an immunofluorescenceassay with monoclonal antibodies directed to measles15 virus. Based upon this diagnosis, oral ribavirin1000mg B.I.D. was given for 14 days. Unfortunately,the patient progressively deteriorated, eventuallydying two months later.In order to ascertain the nature of the20 measles virus present in the patient, reversetranscription and PCR amplification of virus obtainedfrom infected tissues were performed, followed bysequence analysis. The measles virus isolated fromRhesus monkey kidney cells inoculated with tissue from25 this patient's lung biopsy was propagated by two serialpassages in the continuous Vero (monkey kidney) tissueculture cell line. Total infected cell RNA wasextracted at the second Vero cell passage using TRIzolreagent (Life Technologies, Grand Island, NY) according30 to the manufacturer's protocol. Total RNA wassimilarly extracted from the patient's lung biopsymaterial. The measles virus vaccine strain (Moraten)currently used in the United States as a component ofthe trivalent MMR vaccines, was obtained in its35 univalent form (Attenuvaxm, Merck, Sharpe, & Dohme).SUBSTITUTE SHEET (RULE 26)?1015202530W0 98/13501CA 02265554 l999-03- 16PCT/U S97/ 16718This virus was passaged once in Vero cells and totalvaccine infected cellular RNA then was extracted asdescribed above.Each of these RNA preparations was reversetranscribed (RT) to cDNA using random hexameric primersand Maloney murine leukemia virus reverse transcriptase(Perkin—Elmer/Cetus RT—PCR kit reagents, Perkin—Elmer—Cetus, Branchburg, NJ). The cDNA then was amplified byPCR using measles virus-specific oligodeoxynucleotideprimer pairs whose design was based on the Edmonstonmeasles virus sequence described above. These PCRproducts comprised a set of overlapping DNA fragmentsspanning the entire 15,894 nucleotide long measlesgenome. A consensus genomic sequence was establishedby direct analysis of each PCR product, withoutcloning, using the dideoxy terminator cycle-sequencingmethod established by the manufacturer (ABI PRISM 377sequencer and ABI PRISM DNA sequencing kit; Perkin-‘Elmer/Cetus, Foster City, CA). Both strands of thePCR-amplified DNA products were analyzed to eliminatepossible sequencing ambiguities.The nucleotide sequences of selected regionsof the measles virus genomes present in the patient'sviral isolate, as well as in the diseased lung tissue,were compared with that of the Moraten vaccine virus,as well as with the nucleotide sequences of other"measles virus wild-type and vaccine strains. Thissequence analysis revealed identity to the Moratenvaccine strain rather than demonstrating relatedness topast or currently circulating wild-type viruses orother measles vaccine strains.SUBSTITUTE SHEET (RULE 26)I?10CA 02265554 1999-03-16W0 98/13501 PCT/US97/16718- 79 _Example 5ELISA to Detect RSVAn ELISA test is used to detect the presenceof RSV. Peptides are designed and selected based onhomologies to the RSV sequences described herein to bespecific for all subgroup B strains, or for individualwi1d—type, vaccine or revertant RSV subgroup B strainsdescribed herein. These peptides are then coupled toKLH and used to immunize rabbits for the production ofmonospecific polyclonal antibody. A selection of thesepolyclonal antibodies, or a combination of polyclonaland monoclonal antibodies is then used in a "captureELISA" to detect the presence of an RSV antigen.SUBSTHUTESHEET(RULE26)?CA 02265554 l999-03- 16W0 98l1350l PCT/US97l 16718Bibliograghy1. Kapikian, A.Z., et al., Am. J.Epidemo1., gg, 405-421 (1969).2. Chin, J., et al., Am. J. Epidemo1., gg,449-463 (1969).3. Fulginiti, V.A., et al., Am. J.Egidemo1., gg, 435-448 (1969).4. Prince, G.A., et al., J. Virology, §1,721-728 (1986).5. Kim, H.W., et al., Pediatrics, 52, 56-63(1973).6. Hodes, D.S., et al., Proc. Soc. Exp.Biol. Med., 145, 1158-1164 (1974).7. Belshe, R.B., and Hissom, F.K., J. Med.Viro1., 10, 235-242 (1982).8. Black, F.L., et al., Am. J. Epidemiol.,_gg, 442-452 (1986).9. Lennon, J.L., and Black, F.L., Q;Pediatrics, ;Q§, 671-676 (1986).10. Pabst, H.F., et al., Pediatr. Infect.Dis. J., 1;, 525-529 (1992).11. Centers for Disease Control, g?g, gg,369-372 (1991).12. Centers for Disease Control, ggyg,41:S6, 1-12 (1992).13. King, G.E., et al., Pediatr. Infect.Dis. J., 1Q, 883-887 (1991).14. Rota, J.S., et al., Virology, 188, 135-142 (1992).15. Rota, J.S., et al., Virus Res., 3;, 317-330 (1994).16. Lamb, R.A., and Kolakosky, D., pages1177-1204 of Volume 1, Fields Virology, B.N. Fields, etal., Eds. (3rd ed., Raven Press, 1996).SUBST?UTESHEET(RULE26)?W0 98/1350]17.65 (1993).18.6094 (1995).19.5783 (1995).20.9_2.21.702,085.22.WO 96/10400.23.2.1.24.WO 97/06270.25.60/047575.26.WO 97/12032.27.569-579 (1996)28.807 (1995).29.241-256 (1941)30.Sci., USA,No.Virology,263,31.CA 02265554 l999-03- 16PCT/US97/16718Sidhu, M.S., et al., Virology, 193, 50-Garcin, D., et al., EMBO J., ;g, 6087-Radecke, F., et al., EMBO J., 4, 5773-Collins, P.L., et al., Natl. Acad.11563-11567 (1995).Published European Patent ApplicationProc.Published International Application No.Baron, M.D., and Barrett, T., Q;1265-1271 (1997).Published International Application No.U.S. Provisional Patent Application No.Published International Application No.Kato, A., et al., Genes to Cells, 1,Sidhu, M.S., et al., Virology, 208, 800-Shaffer, M.F., et al., J. Immunol., 41,Enders, J.F., et al., N. Engl. J. Med.,153-159 (1960).Enders, J.F., and Peebles, M.E., Proc.Soc. Exp. Biol. Med., §§, 227-286 (1954).32.103,33.Schwarz, A.J.F., Am. J. Dis. Child.,216-219 (1962).Griffin, D.E., and Bellini, W.J., pages1267-1312 of Volume 1, Fields Virology, B.N. Fields, etal., Eds.(3rd ed.,Raven Press, 1996).SUBSTHUTESHEET(RULE26)., ,......_........-............-. _.?CA 02265554 l999-03- 16WO 98/13501 PCTIU S97/ 1671834. Birrer, M.J., et al., Virology, lQ§,381-390 (1981).35. Birrer, M.J., et al., Nature, 29;, 67-69(1981).36. Norby, E., et al., pages 481-507, in IhgParamyxoviruses, D. Kingsbury, Ed. (Plenum Press,1991).37. Peebles, M.E., pages 427-456, in IhgParamyxoviruses, D. Kingsbury, Ed. (Plenum Press,1991).38. Egelman, E.H., et al., J. Virol.,2233-2243 (1989).39. Udem, S.A., et al., J. Virol. Methods,§, 123-136 (1984).40. Udem, S.A., and Cook, K.A., J. Virol.,42. 57-65 (1984).41. Moyer, S.A., and Horikami, S.M., pages3.249-274, in The Paramyxoviruses, D. Kingsbury, Ed.(Plenum Press, 1991).42. Blumberg, B., et al., pages 235-247, inThe Paramyxoviruses, D. Kingsbury, Ed. (Plenum Press,1991).43. Berrett, T., et al., pages 83-102, inThe Paramyxoviruses, D. Kingsbury, Ed. (Plenum Press,1991).3.44. Tordo, N., et al., Sem. in Virology,341-357 (1992).45. Cattaneo, R., et al., EMBO J., §, 681-688 (1987).46. Crowley, J.C., et al., Virology, _§4,498-506 (1988).47. Banerjee, A.K., and Barik, S., et al.,Virology, 188, 417-428 (1992).48. Castaneda, S.J., and Wong, T.C., J;Virol., §;, 2977-2986 (1989).SUBSTITUTE SHEET (RULE 26)I?W0 98/13501CA 02265554 l999-03- 16PCTIU S97/ 16718__33_49. Chan, J., et al., pages 221-231, inGenetics and Pathogenicity of Negative StrandedViruses, B.W.J. Mahy and D. Kolakofsky, Eds. (ElsevierBiomedical Press, 1989).50. Blumberg, B., et al., Cell, 2;, 837-845(1981).51. Blumberg, B., et al., Cell, 32, 559-567(1983).52. Kolakofsky, D., and Blumberg, B.M.,pages 203-213, in Virus Persistence, B.M.J. Mahy, etal., Eds. (Cambridge University Press, 1982).53. Castaneda, S.J., and Wong, T.C., J;Virol., gg, 222-230 (1990).54. Curran, J.A., and Kolakofsky, D.,Virology, 18;, 168-176 (1991).55. Sidhu, M.S., et al., Virology, 19;, 66-72 (1993).56. Sidhu. M.S., et al., Virology, 20;, 631-641 (1994).57. Collins, P.L., et al., pages 1205-1241of Volume 1, Fields Virology, B.N. Fields, et al., Eds.(3rd ed., Raven Press, 1996).58. Crookshanks, F.K., and Belshe, R.B., J;Med. Virol., ;;, 243-249 (1984).59. Crookshanks-Newman, F.K., and Belshe,R.B., J. Med. Virol., ;§. 131-137 (1986).60. Hall, S.L., et al., Virus Res., 2;, 173-184 (1992).61. Karron, R.A., et al., J. Inf. Dis., 112,1445-1450 (1995).62. Anderson, L.J., et al., J. Infect. Dis.,151, 626-633 (1985).63. Collins, P.L., pages 103-162 of EngParamyxoviruses, D.W. Kingsbury, Ed. (Plenum Press, NYand London, 1991).SUBSTWUTESHEETIRULEZB)?CA 02265554 l999-03- 16WO 98/13501 PCT/US97l167 1864. Sullender, W.M., J. Virology, §§, 5425-5434 (1991).65. Lerch, R.A., et al., J. Virology, gg,5559-5569 (1990).66. Mallipeddi, S.K., and Samal, S.K., J;Gen Virol., 14, 2787-2791 (1993).67. Johnson, P.R., et al., J. Virology, §1,3163-3166 (1987).68. Stott, E.J., et al., J. Virology, §;,3855-3861 (1987).69. Henderson, F.W., et al., N. Engl. J.gggg, §_Q, 530-534 (1979).70. Hall, S.L., et al., J. Infect. Dis.,1§;, 693-698 (1991).71. Mufson, M.A., et al., J. Gen. Virol.,é?. 2111-2124 (1985).72. Glezen, W.P., et al., Am. J. Dis._Chi_1;1_-. 14_0. 543-546 (1986).73. Hemming, V.G., et al., Clin. Microbiol.3351: §. 22-33 (1995).74. Collins, P. L. et. al., pages 1313-1351of volume 1, Fields Virology, B. N. Fields, et al.,Eds. (3rd ed., Raven Press, 1996).75. Ling, R., and Pringle, C.R., J. Gen.Virol., 10, 1427-1440 (1989).76. Yu, Q., et al., J. Virology, §§, 2412-2419 (1995).77. McIntosh, K., and Chanock, R.M., pages1045-1072 of Virology, B.N. Fields, et al., Eds. (2nded., Raven Press, 1990).78. Heminway, B.R., et al., page 167 ofAbstracts of the IX International Congress of Virology,P17-2. (1993).79. Mink, M.A., et al., Virology, 185, 615-624 (1991).SUBSTITUTE SHEET (RULE 26)I?CA 02265554 l999-03- 16W0 98/13501 PCTIUS97ll6718-815-80. Dickens, L.E., et al., J. Virology., gg,364-369 (1990).81. Wagner, R.R., and Rose, J.K., pages1121-1135 of volume 1, Fields Virology, B.N. Fields,et al., Eds. (3rd ed., Raven Press, 1996).82. Barik, S., J. Gen. Virol., 1g, 485-490(1993).83. Collins, P.L., et al., pages 259-264 ofVaccines 93: modern approaches to new vaccinesincluding prevention of AIDS, F. Brown et al., Eds.(Cold Spring Harbor Laboratory Press, NY, 1993).84. Kuo, L., et al., J. Virologx., lg, 6892-6901 (1996).85. Huang, Y.T., and Wertz, G.W., glViroloogy, 3;, 150-157 (1982).86. Sambrook, J., et al., Molecular CloniggiA Laboratory Manual, 2nd ed., Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y. (1989).87. Ray, R., et al., J. Virol., gg, 1959-1963 (1995).88. Ray, R., et al., J. Virol., lg, 580-584(1996).89. Stokes, A., et al., Virus Research, lg,43-52 (1993).90. U.S. Patent Application No. 08/059,444. SUBSTITUTE SHEET (RULE 26).,.....—..-....—....-a-....»................» « » -?ICA 02265554 1999-03-16W0 98/ 13501 PCTIUS97/16718SEQUENCE LISTING(1) GENERAL INFORMATION:(i) APPLICANT: Udem, Stephen A.Sidhu, Mohinderjit S.Tatem, Joanne M.Murphy, Brian R.Randolph, Valerie E.(ii) TITLE OF INVENTION: 3' Genomic Promoter Region andPolymerase Gene Mutations Responsible for Attenuation inViruses of the Order Designated Mononegavirales(iii) NUMBER OF SEQUENCES: 79(iv) CORRESPONDENCE ADDRESS:(A) ADDRESSEE: American Home Products Corporation(B) STREET: one Campus Drive(C) CITY: Parsippany(D) STATE: New Jersey(E) COUNTRY: United States(F) ZIP: 07054(V) COMPUTER READABLE FORM:(A) MEDIUM TYPE: Floppy disk(B) COMPUTER: IBM PC compatible(C) OPERATING SYSTEM: PC-DOS/MS-DOS(D) SOFTWARE: Patentln Release #1.0, Version #l.30(vi) CURRENT APPLICATION DATA:(A) APPLICATION NUMBER: Us(B) FILING DATE:(C) CLASSIFICATION:(viii) ATTORNEY/AGENT INFORMATION:(A) NAME: Gordon, Alan M.(B) REGISTRATION NUMBER: 30.637(C) REFERENCE/DOCKET NUMBER: 33,294 PCT(ix) TELECOMUNICATION INFORMATION:(A) TELEPHONE: 973/683-2157(B) TELEFAX: 973/683-4117(2) INFORMATION FOR SEQ ID NO:1:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 15894 base pairs(B) TYPE: nucleic acid(C) STRANDEDNESS: singleSUBSTITUTE SHEET (RULE 26)?.........._.-_ 1W0 98/ 13501CA(D) TOPOLOGY: linear(ii) MOLECULE TYPE: RNA (genomic)(xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:ACCAAACAAA GTTGGGTAAG GATAGATCAA TCAATGATCATCAAGATCCTTAAGGAGCTTGTGGAGCCATTTACCACTCGGCGGGCCCAAGTCAATTGATTCCAGAGTGAATGAGGCGGAGATGGTTCGATGATTCTGGGCAGACACGGCTAGTTGGTGAAGGACCTCTCGAAACAAACCGATTAGCCAGGACTGCATGAAAATGGGGGAGTGCAGGATCACTCCATGGGGGCAAGAGATGTATCACTGCATTATCAGGGAGCATTGTTCCAGAGGAATCATCCAGACTTACTAACAGGGTCAGAGGATCCCAGTCACAACCAATACTTTGAACAAGGAATACCATCCTAAGCTGATTCGATTTAGATTGCTTACGCCGACAGGATTGCTTTTTATCCTGATTTGCTGGTAACTGCACCCATACCCTCTGAGGTTTGAACGGTAAGGAGGCGAGGATGCAACAAGAGCAGAAAAGAAACAAAACACATTACTGGACCGGTGCACTAATAGACCGATGACCTCTGGCCTTATCACATGATGATCTCAGATAGCCCAAATTTGAGCTAAGAAGAGAGAAAATTTCATGGTCGGAAATGATATACTATTAAGTGAGTTATCCATACATGGTAACTCTGGAGCTTTTGGCCGATTCAGCTGGAAAGGCTTGTTTGATTAGGGATAGGACAAACCTTATAGTACCTGGTCAGGTTGTATATTATCCTGACGTTAGCCTTCGCATCATCCAATTAGTTGAAGTGCAGGGTCTTGCTGGTGGAIEAAGGTTGGAIGTCTCTAATCCTGTGACATTGATTGGGATAGACACTTGAGTCTCCTGGAGAAATGCCATGGGCTTACTTTGAAGGTCAGTTCCAGAGATTGC02265554 1999-03-16TATTCTAGTGATCCGAGATGACCCATTACAAATCCCTGGAAATTGGAAACCTTATTTGTGCATAAGGCTGAAGAGGTACCTAGTGATCAAAGACCCTGAGCGCAAAGGCGGTACACCCAAGGTGAGGAACGGATATCAAGTACATATATCAACTATGTATCTTGATGAACCTCAATTCAGAGTAGGAGTGTCCAGCATATCACATTGGCAAATGCATACTSUBSTHUTESHEETKRULEZG). ............a............... VPCT/US97/16718CACTTAGGAT 60GCCACACTTT 120TCAGGATCCG 180GATTCCTCAA 240CCGGATGTGA 300GAGTCTCCAG 360TTAGAGGTTG 420AACATGGAGG 480TCCAGGTTCG 540GGATTCAACA 600GTTACGGCCC 660CAAAGAAGGG 720AGGATTGCCG 780AGAACACCCG 840GTAGAGGCAG 900ccrccwcrwc 960CTTTACCAGC 1020AACAAGTTCA 1080GAACTTGAAA 1140TTTAGATTAG 1200TCTGAACTCG 1260ACTGAGGACA 1320?W0 98/13501AGATCAGTAGGTGAGAATGAGAGAAGCCAGCCCATCTTCCCGCAGGACAGCGGAAGAACAACTAGGTGCGAAAACTTAGGGAGCCGATGGCTCAAGGCCGATATCAGACAGGTCTCAGCACGCGGCCAGGAATCTCCAGGGCGGTTAAGGAGCACCCTCTGATACCGAGGGCTTCTGATGAGAGGCAACAGGTAGGGCCATTTGGAACGGCCCTCGGAACGCCGCACTGAAATAATGAAGAAAACAGCCTCTGCTGTTATAGCGGTTGGAGCTACCGAGAGGAGAGCTACAACCGGCACATCGAAGGTCAAGGCTCAGACAGAGGCCGAGAACCAGGTCCCAGAAGAGCAAGCCCATCGGACCCAGGACAAACCATGCCTGACCTGGAGACATCAAGCACGAATCCAAGACAGGAGGAGAGATATGCTATTTGAAACTGCACTTTCCGAAGCACTTCCGAAGATCGCGTCCATCAGGGCCTACAGGAGTGAAGGGGGAGATGGCCAAAATTGAAGGGAGACACCCAGACAAGTTGGGGGGCAAGAGAAACCGCCCCTAGACAGCTGACGCCCACGGACACCCGACCAGAACAACACAGCCGCGGCACGCCATCTCACTGGCCGGAGCGAGCCCTCAGCAATTGAGCGATGACTGGGTTACAGTGCTGACTCTCAATGAATCTCACTGACCGGAGAAGGAGGGGCTTGGGAAAGACACCCATTTTTATTGACAAGGTGCACCTGACACCCGAACTATTATGATACACGAGGATAGTTGAGTCACCCAAGTATCAGGAAGATAGGGCCCAGCAGTTGACACTGCTGCTTAGGCTCTATAGTGTAACATCCGCCTCAGCCCATCAGTCAAAAACGATCGAGGAAGACCTGCAGGGGGATCAACTGGACGCTGAAATGTTATTATGATCATGGTTCGAAAACAGCGGGATCTGCTCGAGATCCACGACTCTCAATGAAAAAGGGCAGGTGGTGCAAGCGGGGAATGTCTGGTACCAGATGAGCTGTAATCAGAAGAATTAAGAAGC02265554 1999-03-16ATTTCTACACGAGGGTCAAAAGCAAGTGATATCGGAGTCCGCAAGCCATGCAATGACAGAACCCTCCATCACCATCCACTGACTGGAATGCTATGGCAGCAAGAGAAGGCAAGGCGGTGCCTTTGGGAATTTTATGATCAAATCAGGCCTATGTGGATATCCATCTCTATAGCTCCTGAGTTCCTCCGCCCAGACGCGAGCCCAATGTGCTCCCCGAGTGCAATCTCCCCTCTCTGATGTTAATCTCCAAAGATCAACAGSUBSTITUTE SHEET (RULE 25)PCT/US97/16718GGTGATCAAA 1380CAGAGTCGAG 1440GCGAGAGCTG 1500AGCCAAGATC 1560GCAGGAATCT 1620AATCTTCTAG 1680ATTGTTATAA 1740CCCACGATTG 1800CATCCGGGCT 1860ATGGTCAGAA 1920AGGCAGTTCG 1980ACCTCGCATC 2040CCCCCCAAGA 2100CAGCGGTGAA 2160TGATGGTGAT 2220TGGCGAACCT 2280GGGGTTCAGG 2340ACTCCAATCC 2400CCCGGACCCC 2460ATTAGCCTCA 2520TCGAAAGTCA 2580TGTGAGCAAT 2640GAGATCCCAG 2700CCAAGATATT 2760GCTAGAATCA 2820GCAAAATATC 2880?W0 98/13501AGCATATCCAAAGGATCCCAGGCAGAGATTCTCCAAGGAACTAAAGCCGAGCATCACGCACGTTACCTGACAGATGCTGACCAGTCGACCGCCTCCCAAGAAGGGTCGATTCAGAGTCATTGCTGGGGGTCCCTGCCCTTCTGAGCTTGAACAACACCCCTCAACGCAAATCCGTGTTGTGAAGAATGCTGGATTGACAACAACATTTATATTATTGCAAGCACCAGTCTGGTTCAAGAATCTGGAGGAGAAGAATTCCGCCCTGGAAGGACGACCCCACCAGGCCGAGCTGACAAATGGTCGGGAAAAAGTGTAATCCGTGACTCTCCTTGAAGATAATCAACTAGTACTTCCACAATGCGCTCCGATAAGATCCTGGTTGTTGAGGGCAGGTGTTGGCCATAGTTGTTACTAACTCTCCCAAGTGTGCTTATATGAGCGGAATTCAGAGGCGATAGGCGGTCCACATCAATGAAAATCTCACATTAGAGACCTTATGTCAGATGCAAGCATTTACGACCAACACCTCTCATGCAGATGTCACTGGCCGAAACGGACCAGTGATGAGCTCACTCCATTATATGATGATATCAATGAAGTAGAACCTAAATCACAGAGATCTCAACCCACCACTAGGCGACAAGCGATCCCCAGATCCACAGAGACGTACAGCTCACACCTTAATGCGGTTAATCACCCGTCTCGGTCAATGCCTGGGAAGAGGGAACTTCAGAAAAGATGGAGCACAGGCATACCCGCTGAATAGTAAGAAGACGTGATCAAGCATCATGAGAAATCAATCGTTCTCAAGATCCAGAGGACGCCGTCGGGTAAATCCAGCCAAAGGAGCCACTACAGCTCACATTATAAAAACGACTTCGACCTACAGTGAGGAAGGATGATAGGGCCTCCCAAAGCCCGACAGGGCTCAAGGAGAAAGGTATCTGATACCTTTCGGATAACAGTGGCCTTTCATCGACAAGGAGAAAGAAGCCTGGTTTTAGATGAGCAATGGATATCAATCCAGGCAGTTAAATGATGA02265554 1999-03-16TCGCCATTCCCCGACTTGAAAACCCGTTGCAGCTGCTGAATTGTTCCTGAGGCTAGAGGAATGATCTTGCACTTACCTGCAACTTAGGAGCAAGTCGGCATGGCAGGCTGATGCTTTATGAATCGGGCGAAGAACTCCTCTGAAAAACTGCCTAACAACAGCTCGATACCCGGGTATTACCAACCTGCTGTACAGAGCAAGAGTGAAGTCTGCACTTGGTGACTCTCCATTGAAGACCTTTTTGCAGCCACCAAGGACTASUBSTITUTE SHEET (RULE 26)PCT/US97/16718TGGACTTGGG 2940ACCCATCATA 3000CAGCCGACAA 3060GGAATTTCAG 3120CACCGGCCCT 3180GGATCGGAAG 3240CAAGTTCCAC 3300CAACCCCATG 3360CAAAGTGATT 3420TGGGACATCA 3480GTGCCCCAGG 3540TACATGTTTC 3600GCATTTGGGT 3660AAAGAGGCCA 3720GTGTTCTACA 3780GGGAGTGTCT 3840CCGCAGAGGT 3900ACCGTTCCTA 3960GTGACCCTTA 4020CTTCCTGAGG 4080TACTCTGCCG 4140GGGATAGGGG 4200GCACAACTCG 4260AATCGATTAC 4320TCAGTTCCTC 4380TTCAAAGTTC 4440?W0 98/1350]TGTAGACCGTGCCCGGACAAGCCGACGGCACCACCAGCCATGCCCCCGATATTGGAAGGCGACCGAGGTGACTAAACAAACGGCGCCGCGCCCCGGTGCCAATCCAAGACGAGGAAGCCCGGGCCACCAGACCCCAGCCCCGAAGGACCCCTCCTCCTTTCCCACCCCTAGGTCTCAAGGACCGGTCAAAAGCTACAAAGATAACTCTCCACAGTTTTGGCAGAGTGTAGGCCCTAGGCGCTGAACTCTCGAGGCAATCAAGTGCCCAGCAAAAGCCCCCAGCGCGAACACCCCAATCTGCCAAACCACCcccrcccccwACCCAACCGCACTTAGGGCCCCCCCAACCCCACAGGCAGGGGGGGGGCCCACCCACCCCACTCCCAGACTCGATCCGGCGCCGAACCGCATCTCGAAGGGAAGGAGACACTGAACGTCTCTCCATTGGGGTTATGACTCGTCAATAACTGAACCAATTAGCTTCAAGTAGTTGCCACAGCAAGCCATCGAGACAAGCAGGCAAATGCCCGAATCCGAAAGACCCAGGCGGCCCATCCTCCTCAACCGCATCCCTTCCTCAACAGGCATCCGAAAGGAACATACCGACAACCAGACACCAACCCCCCAAAAAACACACGACCACGGCCATCACGGGAGCCACCAAGGACATCAACCAAAAGATCGGGAATCCCTGCCATATTCCAATCTCTCTTTCCAGCCATC#CGAGGGTAAGATGCACTTGAGACACAAGTGCTCAGATACAATCTGAGAGCAGGAGATGAACGACCCCCTCCACGGACCCCAGCACAGAGTGGGACCCCCCACCACCCCACAAGAACTCCTCCCTAGACCACACCCAACGAGGGAGCCCCCCGAACAGAAGGCCCCCAGCGGCAACCAACCCGCAGAAACAACCCGAACGTATCCCACACAATCCACCAAGAATCAAGAATGGCAGTACAAGATAGGGGCAATCATTAGGAGATTGCAGAATGCAATGAAGATTTGCGGACAGCCGGCAGCGAGCCTGGATATTGGCTG02265554 1999-03-16CTCACAATGAAAGCGAGAGGACAGCCCTGACGAGGACCAACGGGAAAGAACACAACCGAAAGATCCTCTCAGAACCCAGACCAACCAATCCCCAGCACCCGGGCCGACAGACCAGAACCCGGAAAGGCCACAGCACCCAAGCCTCTCCAACACCCGACGACTCATCCAATTGTTAACTCTTGGTAGGAATTCATAAAATTAATACAGGAGCCCAGAATATGAGTAGTCCTTTGCACTTCAAAACTACTAATTCAGGGTGTSUBSTHUTESHEET(RULE26)PCT/US97/16718CAGCCAGAAG 4500CCAGCCAGCA 4560CACAAGGCCA 4620CCCCCAAGGC 4680ACCCCCAGCA 4740CCGCACAAGC 4300TCCCCGGCAA 4860ccccGGcccA 4920cccccesrrc 4980AACCATCGAC 5040CCAGCACCGC 5100AGACCACCCT 5160CAACCCGCGC 5220GAGCGATCCC 5230GTCCCCCGGT 5340CACTCAACTC 5400GTCCATCATG 5460CCAGACACCC 5520AGGAAGTGCA 5580AATGCCCAAT 5640ACTACTGAGA 5700AAGACCGGTT 5760GGCAGGTGCG 5820CCAGTCCATG 5880TCAGGCAATT 5940CCAAGACTAC 6000? W0 98/ 13501ATCAATAATGCTCGGGCTCACGGGACCCCAATCAATAAGGAGCAGAGGAAAGTATAGCCTGTCTCGTACACAAGGGTACCGTGTGCAGCCTCCACCAAGTTCACAAGGGAACGATCATTAGTAGTCGAGGTACTTGCACAAATCTGGGGACAGATATTGAGTGTGTCTTGAACAAAAAGGACATCAAAATCACAAGTCTCTATCTCCGGCATCATCCACATCCCAAGGGATTTGCTGGCTCATTAGACTTTCTAGATGTAAGCTGATACCAATTGCTCAGTATCTGCGGATGTTAGAAAATAAAGGCCCGATCCGACGCTACATAGGCTCTTATCTCGAAAAAATGCCTTCCTGTGCTCGACCTAATAGCATCAAGACCCTGAACGGCGTGAATTGACCTATGCAATTGCGGAGTATGAAGAGGGTTGATGAGAACAAGTCCTATGTAAGCTCTTCGTCATTCCCTCTGGATGTCACCACAGTAGGATAGGTTCTGTTTGCATCGGGCAGACTAACTCAACAGTCTATGAACATACTATACAGATATCTATCGCTCGGATACGATAACTCACGTCCGAGATTTCAAGAGTGGTTTTGATGAGGTACCCGATGTACACTCGTACAATTGTGCATGACAAGATCGACCATCCAACGGTCCTCCCTAAGTTGGAGAGGTTTATCGAGGGATCCCCTGGTATGTCAGTCGCTCTGATCAAGCAACCCCGAACAAIAAACGAGACCGTCATTAACAGTCATGTCTCTCCATCTACACTCGAGCATCACAACTATCTTGAAATCCTGTCAGGCTTTGAAGTGGAGGTGGTCGACACAGAAGGGGGTGATATACCACTGTCATCGTGTAAGTCCTCTGCTCCGGGTCTTTCAATCCTTTCTAACATJACAGTCGGGAGCAATATCATTGGGATGCCAAGGAGCACTAGCAGCTTTAATATAGACCAGGCCTCCTCTACAAACCGCACCCATCGGTAGTTAGATAAATGCCAGAACATCTTGAGCTTGATCCGCAGAGATCGGTCAAGGAC02265554 1999-03-16GTGATTTAATCATTATTTGGGCTATGCGCTATTTACTGGGAGTCCTACTTTTGTCCACCGTGCCCAAGTACTTTCATGCCTCCAAGAATGTTGGGAACCGGCAAGTGTTATTGCTGCCGAGGAGGTATCCAGAGGTTGGAAATTGTTGGATAGTCTACATGTTGCTGCAGTAAAGCCTGACTCTTGAAACGCATCAAGCCATTAAAACTTTTCTACAAAGATGATTGATAGGGTTGCTAGCATAAAAGCCGTGCTGACACSUBSHTUTESHEET(RULE26)PCT/US97/16718CGGCCAGAAG 6060CCCCAGCTTA 6120TGGAGGAGAC 6180CATCTTAGAG 6240CATTGTCCTC 6300GCTAGAGGGG 6360TGTTGCAACC 6420AGAGGGGACT 6400CCTCCGGGGG9 6540GTTCATTTTA 6600CACAACAGGA 6660TCACTGCCCG 6720AGACGCTGTG 6780CGTAGGGACA 6840GTCATCGGAC 6900CCTGATTGCA 6960GGGGCGTTGT 7020TCTTACGGGA 7080ACAAATGTCC 7140CACCTGAAAT 7200AGGGTGCAAG 7260ATAACCCCCA 7320GACCTTATGT 7380CCATTGCAGG 7440TCAGCACCAA 7500CACTCTTCAA 7560?W0 98/ 13501AATCATCGGTCATCTCTGACTTGGTGTATCGGCTGCTGAACAATCAGTTCATTCTCAAACATCTATAGTCTAATCTGAGCAGGTGTTATCGCAACCAGTCAGCCCTTTGTCAGCTTCCAGCCCCTTATCATATCGCTGACAATGGAGACACGAGTGGGCAGAGTCTGACACGGTTCAGGGGCCAATGAAGGGTTAGTCCCAACATACCTAACCTGGTCAATGTGGTTTATGCCTATAAAGCTGGTGCCGTTGGGATGGTGGATGAAGTGGAAGATTAAATAACCCGCCAGGAGCTCATGACTAGCTGTCTATGTCGCTGTACTATGACATAGCAAAAGGTAGAAATCCGGAGTAATGATCCACGGGGAAGCTCGTCAAGCACGGATGATCAATCAAGCAATGCTTCCAACCCATTGAAGGGTTGAGCTTAATGGACCTATAACCTAGCCTAACCTCTTCACCTGCGGAGGGATCTCCAATTACGTTTACAGGGGTCCCCACACTTCTGTGGGCATGGGAGCAGCCTGAGGACTCCTTAATCCAGAGAATCAAATGCATTGGTCAAAGGGAAACCCTGTTAGACCCAGGGAATCAGAGTTGTCGTTTGGGGGCTCAGCAACTGATTCTATCACTAGGTGTCTGCAGTGATAGAAATGGGCTGTAGGCGTGTAAATAACAGGATAAATCAAAATACAAATCCAATAGGTGTAATCTGTCCCAATTGGATGGTGAATGTTTTGGCGCCCAGGCCGTCGAATTACATGCTTGCGGATCAGCTGCACACCTCAGAGAGGATAGGGAGATTGGATTATGAACTCAACTCTGCTCAGGGCTTGTATTTAGTATGGGGGAACAACTGAGCTCCGGTGTTCTATGGTGGCTAATTCCCTATGAAATCCCCACAGGCTTTACCCCGACAACAGGGTAAAATCTCCTTCATACTGCTTCGGGACCACAACAATCAACACATTGTAAGGAAGCATGTCAAACTCAACCTACGATCTCATTTTCTAGTGGAATGCCTCAGAATCTAGTCACCCGG02265554 1999-03-16TTCACTGACCTACGACTTCAGATCAATACTCTACTGGAGACCCACTACAAAGTCGAGGTTACTTACCTAGATGTACCGAGCATATGACAATTGGGGGAGCCAGGGATCAGACCGACATGCCTCTCATCTCCGAACAGATGCAAGCACTCTGGGGTCTTGTTTCGGGCCATGTGTATTGGCGAGTGGATACGGCGAAGACTAGTTCCAATCACTTCCAGGGTACTTTTATCTTCACATGGGGGTGGACATAGAAGATGGAASUBSTITUTE SHEET (RULE 26)PCT/US97/16718TAGTGAAATT 7620GAGATCTCAC 7680GTGCAGATGT 7740CCAGAACAAC 7800TCAGAGGTCA 7860ACAATGTGTC 7920TGGAAAAGCC 7980TGTTTGAAGT 8040ACTATCTTGA 8100TCAAACTCGC 8160GGAAAGGTGT 8220AATCCTGGGT 8280ACAGAGGTGT 8340ACAAGTTGCG 8400GCGAGAATCC 8460CTGTTGATCT 8520TGATCACACA 8580TGACTATCCC 8640CGAGATTCAA 8700GCCATGCCCC 8760TGGTGATTCT 8820TTGAACATGC 8880CTTTTAGGTT 8940ACCAAAAACT 9000TCACTCACTC 9060CCAATCGCAG 9120?WO 98/13501ATAGGGCTGCGTGAAATAGACGCTATCTGTATAAGATAGTCTACACTGTGTAAACAATGTCTCATATTCCCGAGGAAGATAGGTTTTCCAAGGACATCAAAGCCCTTTCTCCCATACTTGTGCTAATCTCTGACATTTGACCGCTATGACAACTGATAGATGGAGCCTCTCTTTCCTTAAATGAAGGTACTACATCTGACCAGTAACGGCAGACTCTGATGGCACGGAGGATGCTCAAGCTTGCTGGAGTACCTAAAGGATAGTGAACCACATCAGAATTCAACCAGATCAGCCATCCTGTCAGAACATCGGAAGTTGGGATATCCAAATCCGTGAACTCATGCTTAAGGGGAGAAAGTTGTTTTGGTTTCCATAGGAGGTCGTGACCTTACTGGTTTTGTATTGATGCTTGGTTTCTTCTTCACTTGCTCCACTGCTTTTTATCATGAGAGGGGAGATTTGCTGAAAATGAAAGGTCATCAGTTGGCCATTCAGGTGAAGAAATTTGGCCAAGGCACTTCAATCACATGATAAGAAAAACGTTATACCCTGGAGTATGCTCAAGCACCGCCAATGTCATCATGTAATCAGGCTCAAAAAGGGACACTAACTATTAACTTGGACAGTCAAGAAGACACACACGTTGCTATAAATGTATTGTGAGGTATACAGCCTGCACTCGTACCTGCAGCACTGAAATACTTAATTGAAGTTCTCATTTTGTTAGGAAATGCCATATTTTCCGCTGACCCGGGTTAACACTGCTTTATGCGCTGCTCTCCGTCACCCAGATAGGGTCCAAAAGTTCACCTGAGTCCCTCATAAAAAACGGAGTCCAAGCTATTTATTTAAGGAATTCGCTCACGGCTTGGGAGTTTACATCTGAGATGAGCTGTATTCTTTCAGTAAAGAATGTCATAGAAGCTTCTAGGGGAATCCAACTGAGGGILTATATGATGTTCTCTCTAGATTATCAGAAGTTTACATGAATCAGTGGAATCATTCCCCCTGCAATGAGCAGTGCTCTTAGCCTAAAGGGAATG02265554 1999-03-16CATCAGGCATGTGGTTCCCCAGATAGCCCGCGCTTACAGCATTTTCCAACTAGGAGTTATCATAGAAGACGTACTCCAAACCTAGGCTCCGCACAGCTCCGTCAGTGATTCACTGGTAGTGTCTCAACATGGGGAGGTTAAAGAGTCAGATTATCAAATTAACAGTAGAATGACCAAAACCATTTTCATACGGCCACCCCGCCTAAAGTCAATCAACGGCTGCTGCAGACCGTTGATAACGGATAGTGATGGATTCAGTTSUBSTITUTE SHEET (RULE 26)PCT/US97l16718ACCCACTAGT 9180GTTATGGACT 9240ATAGTTACCA 9300CTGGAGGACC 9360CAAATGATTA 9420CCGGCCCACT 9480AAAGAGTCAA 9540GTCAGTGATA 9600GAATTGAGGG 9660CAGTGGTTTG 9720AAATCACAAA 9780TCAGTTGAGT 9840GTATATTACC 9900ATGACAGAGA 9960TACATGTGGA 10020GTAGCCATGC 10080CTCAGAGGTG 10140GGGTTTTCTG 10200ACTGATGACA 10260AGACTTGAAG 10320ATTGTGTATG 10380TATCGTGACA 10440ACAATCCGGA 10500TGGAAATCTT 10560CTGACAATGT 10620TACCCGAAAG 10680?W0 98/13501AGTTCCTGCGTTAATGATTCTCCATGACCCGTAGACTTTTTAATCTCAAAATTTGACTAAGTCACAGGGGGGAACGTGAGACACTGATCAATCTCAAGAATAAATGAGATCTGTCCTGTAATAAAGTCCCGTCAGAAGCTGAGTAAGGATTACCCAGCACACTTTGTAATCAATTGTTTCTGTCCCAATCAAACAAGGGCATGACCGTTACTCTTGGCTTACAACGACCTTGAATATGAGATCTCAAGAGCACAACAACCTTACGACCCTGAGCTTTGACTGAGTTCAACTGCTAAAATGCGGGATTGGCGGCACTCCACGGGGCCAGTCAGCAGCAAAATCCGGAGAATGTACTGCCTTTTACGGATTGTGTAAGTGACCAATGATCAAGTGGACCATCTGCTTCGTTAATGGCCCTACTCTTAGGCAAATCACATTTTACTCAAGAGCAGCATGCAGTCCTTGCATATCACAATCAATCTTAATAAGGCAGGCTGTTTAATGATTCTCGGGGGACTCTCACCCAAGGGAACCATATGATGCTGTCTTACAACTTACAAAAAAATATTTTAACTCTAGCTGTTAAAAACCTGGGTTTATAGATGGAAGCTTAATTGGAGATCCCTCATTTTCCTCATTGCCATCTTCATTAAGCACCATTCGTGCAAGGGGAACCTTAAGAAGGCTACATGTTTGTCTATTATCGCAAGATAATATTGCTATCCCTGAACGTCAACCATGAATGGCACTGTGTCAGAAACAGCCTCACTAATCATTCCTAGCCGGGTCACGTGATAATGTAGCCTGAAAGATGAGGGCATGAGGACAATGGTCTCAGGAGTACTCCCGAAGGGTTCCCTCAACGAGACAGTATGAGACCATTCCAGTGGCTCCCCCGACCTAGTACCCTATCCTATCTATAACAATCAGACAACGGGAAGCATATTGGCCACAAAAGGAATGTGTATTCTGCAACAATGGCTCCTAAAAGTCCCGGGATGTTGCCCGCTCCTCGGTGATCCTGCCTGAAGAACTGGGCTAG02265554 1999-03-16GAGGCTTGTATGTTGTAAGTAAAGGAGATCCCAAGTGATTGATGGCCAAGCCCCAAAGATCCCAGTCCACAGTAATTCGGCAGTGCATTTCAGCTTGTTTGCATAAGAGGTGACGCCCATGGGAGGTATACCTGGCTGCTCATAGCCGTATGCTAGAGTATCACCTCAAGATATTATGATGTCAGAGACTTAAAAGCATCGATACAGCAAAGTCATACCCTATTGGGGGGAGTAACATCAGACCCTCCATCGACCCTTACSUBSTITUTE SHEET (RULE 26)1PCT/U S971 16718GATGTTTTCC 10740GGAGCTTACC 10800AAGGAAACAG 10860GCTGAAAATC 10920GATGAGCACG 10980CTCAAAGAAA 11040ACAAGTACCA 11100CAGGACCAAG 11160ATCACGACTG 11220GCACAGAGGC 11280CTTGAGACCT 11340ATCCCGTTAT 11400GAAGGGTATT 11460TATGAGAGCG 11520ACAAAAAGGG 11580ACTAGAGATT 11640GCAAATGAGA 11700GGGCTACTTG 11760ATAGTTGATG 11820GAGAGAGGTT 11880ATTCTGATCT 11940CTCCTCACAA 12000ATGAATTATC 12060TCAATTGCTG 12120CAAGTAATGA 12180TCAGCAAATC 12240?W0 98/13501TTGTATGTGTTCCATAGTCCAGGGACTGGCTCCTGGATCAAAGGCCTGATTGTCCAATTAGAAATGTCCTATATGTGGGCTAGAATCTATGATCAGTCAAAGGAAACATCTGAAGCTTGCCAGTGTACTCCTAGGCAAAGCGACTAATTTCCCTTGTCCGCAGATAAGAATTTTAGAAACTTCACGTCGACCCGCAAGCTCTTTAATTGAAATTTGTTACCTATGATTGATAGGGGATGATCACTATCTAGACCATCAGGCCAGAGCATCAAACCCAATGGGCATTCCTCTAGTGTCACATCGAGCCAGCTGACTATGAACATTGACAAAGAGGCTAGCTGCGAGGCCACCTACGGATGGATCCTTGAGACTTCGTAAGAATGGGCTTACGGCCAATGTGAGCGCATAGGAGTGGCGAGGGGTTGATACTATTGTTTCGAAACAGATTGTAGAGCTGAGGCAGAGATGCAATGGTCCACACCTGGTAACACGATATCAATCTTGGGCCAGGAAATATCAGCAACTAGACTCCTTAAAAGGATATGGACAGGCGGGGCAAGAGATGAGGAAGGCAATTCAGAGGAGTCATGTTCGAGGACGGCcrwnrrcsscTTTTTTGTCCGTCCCATATAGCCCCAAGTCGGTGATGATGAGCCTGGAGGTTGAGGGATCTATACCACAAAACTTTATATCTCGAGAAAGTGCGTGATCCGCAGAGCTATACAAGGCTATCCCCAACTATAAATTTGAGAAGTTTCATAATGTGCGGCCAATGGGTGAGCTCAAGAACATTATTCCATGAATATTATAGTAGTCTATTGCGGGGGTTAACCAGGGATGGTCAGTGCAGCTCTATTTACGGGTCATGAGACCCTCGGGTTGTTGGTTCTACGATCCTTGCGATAGCTCTTGAGCTAAGGGTGTAGCACTCATCTCCAACGAACCAACAAGGATACCGGATCCGATGATAGAGTACCAACCCACACCCAGAGATCACATTTTAGGACCAENTCTGAGTTTCTTCAATTGGGCTGTTGTCATC02265554 1999-03-16AACTGCAAGGTGACAGTAAAACCTAGGGCAAGGCATGCTGCTCTCGAGTGGCTATTGACAGGCGAGAGCTCCTTGAGGTCATGTGTCATCCCAACTGGATCACTGATGAGATCTGCTGTTGAACGAAGCCGATCACTCCCAGTGAAATACCAATCTCTCAAATGCTTCTAATCTAACACGTCATCCCAGGATTGATATATCCATAGGAGGAGCTAAGTCCGAATGAAATTGCTCATAGAGATTTGATGTAGTTCCTTTCTSUBSTITUTE SHEET (FIULE 26)PCT/US97/16718TTTGTCCTGA 12300GAAGAGGACG 12360GCTCATGAAA 12420GATACCACAA 12480ATAACCAGAT 12540GGAAGAAAGA 12600CTAAGAAGCC 12660CCTGATGTAC 12720TGCGAGTGTG 12780GATATTGACA 12840AGAACAGACA 12900AGAATAGCAA 12960TGGTTGTTGG 13020ATCTCAACTT 13080TCAGGTACAT 13140TTTGTCATAT 13200GGGTTGGGTG 13260GTATTACATC 13320ATACCCAGCT 13380GATAATGCAC 13440CACCTTGTGG 13500ACAGCACTAT 13560TCAGCTCTCA 13620CCAAGATTAT 13680CATTATCATA 13740AGAATGAGCA 13800?WO 98113501AAGGAGTGTTGGCATTGTGGCAACTGTGTGAAGAGTTAGAGATTCGACAAGGACCTGCCCATATCAAGGCTTGTAGACCAGATTGAGAGTCAAAGATCGGATGATGTTGCGGGGCAATCTCTTGCTACAAACGGCTTGTTAACTAAACAAAATTAGCACCTTGTCAAAGTTCAATTTCATAGACCTTGCCTGGCTCTGCTGGGATTTTGTTATACCCTAGAGGCTAACCGGGACTTCACCCAATTGTGGGCTATAGAGCATAAGGTGCTTTATTATAGAGCAACATGGTTAGAGTTCACACATCCAGGCAACCAATTCGAAGAGGCTAGGTTACTCATGCTGATCCAGGACAGCAACAACAAAATTGCTCCGCCAATTATAGCTGTTGAGCTTGGGTGAGGTGCTTCTATCTATCCCTCCGCTCTTTAACAGTTAGTAATTAACAAAGATCCTGGGCAAATCAGGGATTTATACAGCAACGCTAATGAATTGGACTTATAAGACGCAGTTGGTGCTGATCCAGTCAATGCTCCCTATCCATGTACACATGCTTTTCTCTTGTAAACACTTATGGTCTAAGACTTATCTCCAGTCTCTGACTTTTCATTTTCGATCTCAAATAAAAGATATCAGAAATCCATGATATCAACATGGATCGGGTTAATAGTGGGGGAAGTTGGCCGGGAGGCCCGATCCCTACCTACTATAGAGAATAGGATCAAATAAGTTATGTTCATATCTACCTGAAAAGAGGTCACATCCAGTAGAGGTGAATTGCGGGTTAAGCCACCCGTCCTTCACTATATGACCTAGTGAAAGCGAGTGTTCTGGCCGGTAGAGAACAGGATCTTCATCTCCGGCGACGCCCTCGCTGAGCATCAAACACAAGCAACTTTCCGCAGTAATTAGGAGCTATGTTGATTTTCCGCCAATTGTCGAACAAAGTCACGTGCTAGTGTGGGAGCTAGAGGATACTGGTGATTAGGGTCTCACTGAATCTTATTAAGCAGCATATCCATTAAATATCAATCCTGGCAATTAA02265554 1999-03-16AAAGATCTACTGATGCTCAACCTCQ§CCTGCGAGGATGTAAGATTTGTACATGTGCAGTTGTGGAACATAAGGATCGATCTGAGGTAAATGGATTTCAGAGCACAATCTTAATCGGGTTGATGCCTTGAGCACTTATAAGTTCTAGATCTCAGAATGGGAGGTAGGCAGTGTTTATCCATATTGGCAGCCTAAGCTTATGTTATAGAGAATTTGGTTATGGATAATTGAAGCAACTAAGCTACTCTGAAACGGACCTAAGSUBSTITUTE SHEET (RULE 26)PCTIU S97/ 16718AAGAAATTCT 13860AACTTGCACA 13920TTGTTGAATG 13980GTACCGGACA 14040TGTCAACCAG 14100CTAACCGACC 14160AATCCAATTA 14220AAACAGATAA 14280GTCAGTCAGC 14340CCCCCACACG 14400CCCATTTCAG 14460AACTCATCTG 14520CCAGGGGAAG 14580GAGATACTTA 14640GGTCAAAGGG 14700GTAGGTAATA 14760GTAGATTGCT 14820TCAGATATAG 14880ATCTTATCGA 14940CCTTTCAGCG 15000GTGAACCTTG 15060ACAGATCTCA 15120TCATCTGTGA 15180TGCATACAAG 15240AAACTTACAC 15300CTGTGCAAAG 15360?GCTTAATTCTTCAACAAGGGATCTAGGATCGATAAATAAGTATCTTCGTTACGTGAGTGGATACAGTGCCTGGTTAGGCACAGCTTTGTCTyrAlaProPro GluIle LeuThr LeuPCT/US97/16718ATACTCATCCATGTTCCACGACCCGCAAATTTTATCCAGAAAGAATCTATGTTTTTAAGGCTGATTAAGGTTATTTGCAATGGTVal His15Glu30TyrCys Gln45Asn60Lys75AsnCA 02265554 1999-03-16W0 98/ 1 3501- 97 _AATTGATCCA CCATGATGTT GCCTCAGGGC AAGATGGATTTCTACAGGGA GTTGGCAAGA TTCAAAGACA ACCAAAGAAGCTTACCCCGT ATTGGTAAGT AGCAGGCAAC GAGAACTTATTTTGGGGGCA CATTCTTCTT TACTCCGGGA ACAGAAAGTTATCTCAAGTC CGGCTATCTG ATACTAGACT TACACCAGAACCAAGTCAGA GAAACAGATT ATTATGACGG GGGGTTTGAATAACAGTCAA GGAGACCAAA GAATGGTATA AGTTAGTCGGACTAATTGGT TGAACTCCGG AACCCTAATC CTGCCCTAGGTATATTAAAG AAAACTTTGA AAATACGAAG TTTCTATTCC(2) INFORMATION FOR SEQ ID NO:2:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 2183 amino acids(B) TYPE: amino acid(C) STRANDEDNESS:(D) TOPOLOGY: linear(ii) MOLECULE TYPE: protein(xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:Met Asp Ser Leu Ser Val Asn Gln Ile Leu1 5 10Asp Ser Pro Ile Val Thr Asn Lys Ile Val20 25Arg Val Pro His Ala Tyr Ser Leu Glu Asp35 40Ile Lys His Arg Leu Lys Asn Gly Phe Ser50 55Asn Val Glu Val Gly Asn Val Ile Lys Ser65 70Ala His Ser His Ile Pro Tyr Pro Asn Cys85 90SUBSTITUTE SHEET (RULE 26) Gln MetLeu ArgGln AspIle IleSerTyrPhe95LeuLeuAlaAsnAsnPro80Aan154201548015540156001566015720157801584015894?CA 02265554 1999-03-16W0 98/13501 PCTIUS97/16718Ile Glu Asp Lys Glu Ser Thr Arg Lys Ile Arg Glu Leu Leu Lys Lys100 105 110Gly Asn Ser Leu Tyr Ser Lys Val Ser Asp Lys Val Phe Gln Cys Leu115 120 125Arg Asp Thr Asn Ser Arg Leu Gly Leu Gly Ser Glu Leu Arg Glu Asp130 135 140Ile Lys Glu Lys Val Ile Asn Leu Gly Val Tyr Met His Ser Ser Gln145 150 155 160Trp Phe Glu Pro Phe Leu Phe Trp Phe Thr Val Lys Thr Glu Met Arg165 170 175Ser Val Ile Lys Ser Gln Thr His Thr Cys His Arg Arg Arg His Thr180 185 190Pro Val Phe Phe Thr Gly Ser Ser Val Glu Leu Leu Ile Ser Arg Asp195 200 205Leu Val Ala Ile Ile Ser Lys Glu Ser Gln His Val Tyr Tyr Leu Thr210 215 220Phe Glu Leu Val Leu Met Tyr Cys Asp Val Ile Glu Gly Arg Leu Met225 230 235 240Thr Glu Thr Ala Met Thr Ile Asp Ala Arg Tyr Thr Glu Leu Leu Gly245 250 255Arg Val Arg Tyr Met Trp Lys Leu Ile Asp Gly Phe Phe Pro Ala Leu260 265 270Gly Asn Pro Thr Tyr Gln Ile Val Ala Met Leu Glu Pro Leu Ser Leu275 280 285Ala Tyr Leu Gln Leu Arg Asp Ile Thr Val Glu Leu Arg Gly Ala Phe290 295 300Leu Asn His Cys Phe Thr Glu Ile His Asp Val Leu Asp Gln Asn Gly305 310 315 320Phe Ser Asp Glu Gly Thr Tyr His Glu Leu Ile Glu Ala Leu Asp Tyr325 330 335Ile Phe Ile Thr Asp Asp Ile His Leu Thr Gly Glu Ile Phe Ser Phe340 345 350Phe Arg Ser Phe Gly His Pro Arg Leu Glu Ala Val Thr Ala Ala Glu355 360 365Asn Val Arg Lys Tyr Met Asn Gln Pro Lys Val Ile Val Tyr Glu ThrSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/13501 PCT/US97/16718370 375 380Leu Met Lys Gly His Ala Ile Phe Cys Gly Ile Ile Ile Asn Gly Tyr385 390 395 400Arg Asp Arg His Gly Gly Ser Trp Pro Pro Leu Thr Leu Pro Leu His405 410 415Ala Ala Asp Thr Ile Arg Asn Ala Gln Ala Ser Gly Glu Gly Leu Thr420 425 430His Glu Gln Cys Val Asp Asn Trp Lys Ser Phe Ala Gly Val Lys Phe435 440 445Gly Cys Phe Met Pro Leu Ser Leu Asp Ser Asp Leu Thr Met Tyr Leu450 455 460Lys Asp Lys Ala Leu Ala Ala Leu Gln Arg Glu Trp Asp Ser Val Tyr465 470 475 480Pro Lys Glu Phe Leu Arg Tyr Asp Pro Pro Lys Gly Thr Gly Ser Arg485 490 495Arg Leu Val Asp Val Phe Leu Asn Asp Ser Ser Phe Asp Pro Tyr Asp500 505 510Val Ile Met Tyr Val Val Ser Gly Ala Tyr Leu His Asp Pro Glu Phe515 520 525Asn Leu Ser Tyr Ser Leu Lys Glu Lys Glu Ile Lys Glu Thr Gly Arg530 535 540Leu Phe Ala Lys Met Thr Tyr Lys Met Arg Ala Cys Gln Val Ile Ala545 550 555 560Glu Asn Leu Ile Ser Asn Gly Ile Gly Lys Tyr Phe Lys Asp Asn Gly565 570 575Met Ala Lys Asp Glu His Asp Leu Thr Lys Ala Leu His Thr Leu Ala580 585 590Val Ser Gly Val Pro Lys Asp Leu Lys Glu Ser His Arg Gly Gly Pro595 600 605Val Leu Lys Thr Tyr Ser Arg Ser Pro Val His Thr Ser Thr Arg Asn610 615 620Val Arg Ala Ala Lys Gly Phe Ile Gly Phe Pro Gln Val Ile Arg Gln625 630 635 640Asp Gln Asp Thr Asp His Pro Glu Asn Met Glu Ala Tyr Glu Thr Val645 650 655SUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/ 13501 PCT/US97/16718-100-Ser Ala Phe Ile Thr Thr Asp Leu Lys Lys Tyr Cys Leu Asn Trp Arg660 665 670Tyr Glu Thr Ile Ser Leu Phe Ala Gln Arg Leu Asn Glu Ile Tyr Gly675 680 685Leu Pro Ser Phe Phe Gln Trp Leu His Lys Arg Leu Glu Thr Ser Val690 695 700Leu Tyr Val Ser Asp Pro His Cys Pro Pro Asp Leu Asp Ala His Ile705 710 715 720Pro Leu Tyr Lys Val Pro Asn Asp Gln Ile Phe Ile Lys Tyr Pro Met725 730 735Gly Gly Ile Glu Gly Tyr Cys Gln Lys Leu Trp Thr Ile Ser Thr Ile740 745 750Pro Tyr Leu Tyr Leu Ala Ala Tyr Glu Ser Gly Val Arg Ile Ala Ser755 760 765Leu Val Gln Gly Asp Asn Gln Thr Ile Ala Val Thr Lys Arg Val Pro770 775 780Ser Thr Trp Pro Tyr Asn Leu Lys Lys Arg Glu Ala Ala Arg Val Thr785 790 795 800Arg Asp Tyr Phe Val Ile Leu Arg Gln Arg Leu His Asp Ile Gly His805 810 815His Leu Lys Ala Asn Glu Thr Ile Val Ser Ser His Phe Phe Val Tyr820 825 830Ser Lys Gly Ile Tyr Tyr Asp Gly Leu Leu Val Ser Gln Ser Leu Lys835 840 845Ser Ile Ala Arg Cys Val Phe Trp Ser Glu Thr Ile Val Asp Glu Thr850 855 860Arg Ala Ala Cys Ser Asn Ile Ala Thr Thr Met Ala Lys Ser Ile Glu865 870 875 880Arg Gly Tyr Asp Arg Tyr Leu Ala Tyr Ser Leu Asn Val Leu Lys Val885 890 895Ile Gln Gln Ile Leu Ile Ser Leu Gly Phe Thr Ile Asn Ser Thr Met900 905 910Thr Arg Asp Val Val Ile Pro Leu Leu Thr Asn Asn Asp Leu Leu Ile915 920 925SUBSTITUTE SHEET (RULE 26)?CAW0 98ll3501- 101 —Met Ala Leu Leu930Pro Ala Pro Ile935Arg GlyMet945Val950Ser Arg Leu Phe Arg Asn Ile GlyIle Ala Met Ile Leu Ala970Asp Leu Lys965ArgThr Gln Val980Met Thr Gln Gln985Leu His ProAla995Asp Trp Ser Asp Pro Tyr Ser Ala1000AsnIle Thr Arg Leu1010Asn Ile Thr1015Leu Lys AlaSer Pro Asn Pro Met Phe1025Leu Lys Gly Leu1030Glu Asp Glu Leu Ala Ala Phe Leu1045Gly MetPro Arg Ala Ala His Glu Ile Leu Asp His1060 1065Glu Ser Ile Ala Gly Met Leu Asp Thr Thr1075 1080Ser Met Arg Lys Gly Gly Leu Thr Ser Arg1090 1095105002265554 1999-03-16PCTIUS97/16718Gly Met940Asn Tyr Leu AsnPro Val Thr Ser960Asp Ser955Met Pro Glu975Ser Leu GluSer Ser Phe990Gly Asp LeuLeu Val Cys Val Gln1005SerArg Phe Val Leu Ile His1020Glu1040His Asp Asp Ser1035LyeIle Val1055Asp Arg His IleSer Val Thr Gly Ala Arg1070Lys Gly Leu Ile Arg Ala1085Val Ile Thr Arg Leu Ser1100Asn Tyr Asp Tyr Glu Gln Phe Arg Ala Gly Met Val Leu Leu Thr Gly1105 1110Arg Lys Arg Asn Val Leu Ile Asp Lys Glu1125Ala Arg Ala Leu Arg Ser His Met Trp Ala1140 1145Pro Ile Tyr Gly Leu Glu Val Pro Asp Val1155 1160His Leu Ile Arg Arg His Glu Thr Cys Val1170 1175Val Asn Tyr Gly Trp Phe Phe Val Pro Ser1185 1190Ile Asp Lys Glu Thr Ser Ser Leu Arg Val11301115 1120Ser Cys Ser Val Gln Leu1135Arg Leu Ala Arg Gly Arg1150Leu Glu Ser Met Arg Gly1165Ile Cys Glu Cys Gly Ser1180Gly Cys Gln Leu Asp Asp1195 1200Pro Tyr Ile Gly Ser ThrSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/ 13501 PCTIUS97/16718-102-1205 1210 1215Thr Asp Glu Arg Thr Asp Met Lys Leu Ala Phe Val Arg Ala Pro Ser1220 1225 1230Arg Ser Leu Arg Ser Ala Val Arg Ile Ala Thr Val Tyr Ser Trp Ala1235 1240 1245Tyr Gly Asp Asp Asp Ser Ser Trp Asn Glu Ala Trp Leu Leu Ala Arg1250 1255 1260Gln Arg Ala Asn Val Ser Leu Glu Glu Leu Arg Val Ile Thr Pro Ile1265 1270 1275 1280Ser Thr Ser Thr Asn Leu Ala His Arg Leu Arg Asp Arg Ser Thr Gln1285 1290 1295Val Lys Tyr Ser Gly Thr Ser Leu Val Arg Val Ala Arg Tyr Thr Thr1300 1305 1310Ile Ser Asn Asp Asn Leu Ser Phe Val Ile Ser Asp Lys Lys Val Asp1315 1320 1325Thr Asn Phe Ile Tyr Gln Gln Gly Met Leu Leu Gly Leu Gly Val Leu1330 1335 1340Glu Thr Leu Phe Arg Leu Glu Lys Asp Thr Gly Ser Ser Asn Thr Val1345 1350 1355 1360Leu His Leu His Val Glu Thr Asp Cys Cys Val Ile Pro Met Ile Asp1365 1370 1375His Pro Arg Ile Pro Ser Ser Arg Lys Leu Glu Leu Arg Ala Glu Leu1380 1385 1390Cys Thr Asn Pro Leu Ile Tyr Asp Asn Ala Pro Leu Ile Asp Arg Asp1395 1400 1405Ala Thr Arg Leu Tyr Thr Gln Ser His Arg Arg His Leu Val Glu Phe1410 1415 1420Val Thr Trp Ser Thr Pro Gln Leu Tyr His Ile Leu Ala Lys Ser Thr1425 1430 1435 1440Ala Leu Ser Met Ile Asp Leu Val Thr Lys Phe Glu Lys Asp His Met1445 1450 1455Asn Glu Ile Ser Ala Leu Ile Gly Asp Asp Asp Ile Asn Ser Phe Ile1460 1465 1470Thr Glu Phe Leu Leu Ile Glu Pro Arg Leu Phe Thr Ile Tyr Leu Gly1475 1480 1485SUBSTITUTE SHEET (RULE 26)?W0 98/ 13501CA 02265554 1999-03-16PCT/US97/16718-103-Gln Cys Ala Ala Ile Asn Trp Ala Phe Asp Val His Tyr His Arg Pro1490 1495 1500Ser Gly Lys Tyr Gln Met Gly Glu Leu Leu Ser Ser Phe Leu Ser Arg1505 1510 1515 1520Met Ser Lys Gly Val Phe Lys Val Leu Val Asn Ala Leu Ser His Pro1525 1530 1535Lys Ile Tyr Lys Lys Phe Trp His Cys Gly Ile Ile Glu Pro Ile His1540 1545 1550Gly Pro Ser Leu Asp Ala Gln Asn Leu His Thr Thr Val Cys Asn Met1555 1560 1565Val Tyr Thr Cys Tyr Met Thr Tyr Leu Asp Leu Leu Leu Asn Glu Glu1570 1575 1580Leu Glu Glu Phe Thr Phe Leu Leu Cys Glu Ser Asp Glu Asp Val Val1585 1590 1595 1600Pro Asp Arg Phe Asp Asn Ile Gln Ala Lys His Leu Cys Val Leu Ala1605 1610 1615Asp Leu Tyr Cys Gln Pro Gly Thr Cys Pro Pro Ile Arg Gly Leu Arg1620 1625 1630Pro Val Glu Lys Cys Ala Val Leu Thr Asp His Ile Lys Ala Glu Ala1635 1640 1645Arg Leu Ser Pro Ala Gly Ser Ser Trp Asn Ile Asn Pro Ile Ile Val1650 1655 1660Asp His Tyr Ser Cys Ser Leu Thr Tyr Leu Arg Arg Gly Ser Ile Lys1665 1670 1675 1680Gln Ile Arg Leu Arg Val Asp Pro Gly Phe Ile Phe Asp Ala Leu Ala1685 1690 1695Glu Val Asn Val Ser Gln Pro Lys Ile Gly Ser Asn Asn Ile Ser Asn1700 1705 1710Met Ser Ile Lys Asp Phe Arg Pro Pro His Asp Asp Val Ala Lys Leu1715 1720 1725Leu Lys Asp Ile Asn Thr Ser Lys His Asn Leu Pro Ile Ser Gly Gly1730 1735 1740Asn Leu Ala Asn Tyr Glu Ile His Ala Phe Arg Arg Ile Gly Leu Asn1745 1750 1755 1760SUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/13501 PCT/US97/16718— 104 -Ser Ser Ala Cys Tyr Lys Ala Val Glu Ile Ser Thr Leu Ile Arg Arg1765 1770 1775Cys Leu Glu Pro Gly Glu Asp Gly Leu Phe Leu Gly Glu Gly Ser Gly1780 1785 1790Ser Met Leu Ile Thr Tyr Lys Glu Ile Leu Lye Leu Asn Lys Cys Phe1795 1800 1805Tyr Asn Ser Gly Val Ser Ala Asn Ser Arg Ser Gly Gln Arg Glu Leu1810 1815 1820Ala Pro Tyr Pro Ser Glu Val Gly Leu Val Glu His Arg Met Gly Val1825 1830 1835 1840Gly Asn Ile Val Lys Val Leu Phe Asn Gly Arg Pro Glu Val Thr Trp1845 1850 1855Val Gly Ser Val Asp Cys Phe Asn Phe Ile Val Ser Asn Ile Pro Thr1860 1865 1870Ser Ser Val Gly Phe Ile His Ser Asp Ile Glu Thr Leu Pro Asn Lys1875 1880 1885Asp Thr Ile Glu Lys Leu Glu Glu Leu Ala Ala Ile Leu Ser Met Ala1890 1895 1900Leu Leu Leu Gly Lys Ile Gly Ser Ile Leu Val Ile Lys Leu Met Pro1905 1910 1915 1920Phe Ser Gly Asp Phe Val Gln Gly Phe Ile Ser Tyr Val Gly Ser His1925 1930 1935Tyr Arg Glu Val Asn Leu Val Tyr Pro Arg Tyr Ser Asn Phe Ile Ser1940 1945 1950Thr Glu Ser Tyr Leu Val Met Thr Asp Leu Lys Ala Asn Arg Leu Met1955 1960 1965Asn Pro Glu Lys Ile Lys Gln Gln Ile Ile Glu Ser Ser Val Arg Thr1970 1975 1980Ser Pro Gly Leu Ile Gly His Ile Leu Ser Ile Lys Gln Leu Ser Cys1985 1990 1995 2000Ile Gln Ala Ile Val Gly Asp Ala Val Ser Arg Gly Asp Ile Asn Pro2005 2010 2015Thr Leu Lys Lys Leu Thr Pro Ile Glu Gln Val Leu Ile Asn Cys Gly2020 2025 2030Leu Ala Ile Asn Gly Pro Lys Leu Cys Lys Glu Leu Ile His His AspSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/ 13501 PCT/US97/16718-105-2035 2040 2045Val Ala Ser Gly Gln Asp Gly Leu Leu Asn Ser Ile Leu Ile Leu Tyr2050 2055 2060Arg Glu Leu Ala Arg Phe Lys Asp Asn Gln Arg Ser Gln Gln Gly Met2065 2070 2075 2080Phe His Ala Tyr Pro Val Leu Val Ser Ser Arg Gln Arg Glu Leu Ile2085 2090 2095Ser Arg Ile Thr Arg Lys Phe Trp Gly His Ile Leu Leu Tyr Ser Gly2100 2105 2110Asn Arg Lys Leu Ile Asn Lys Phe Ile Gln Asn Leu Lys Ser Gly Tyr2115 2120 2125Leu Ile Leu Asp Leu His Gln Asn Ile Phe Val Lys Asn Leu Ser Lys2130 2135 2140Ser Glu Lys Gln Ile Ile Met Thr Gly Gly Leu Lys Arg Glu Trp Val2145 2150 2155 2160Phe Lys Val Thr Val Lys Glu Thr Lye Glu Trp Tyr Lys Leu Val Gly2165 2170 2175Tyr Ser Ala Leu Ile Lys Asp2180(2) INFORMATION FOR SEQ ID NO:3:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 15894 base pairs(B) TYPE: nucleic acid(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: DNA (genomic)(xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:ACCAAACAAA GTTGGGTAAG GATAGATCAA TCAATGATCA TATTCTAGTA CACTTAGGAT 60TCAAGATCCT ATTATCAGGG ACAAGAGCAG GATTAGGGAT ATCCGAGATG GCCACACTTC 120TAAGGAGCTT AGCATTGTTC AAAAGAAACA AGGACAAACC ACCCATTACA TCAGGATCCG 180GTGGAGCCAT CAGAGGAATC AAACACATTA TTATAGTACC AATCCCGGGA GATTCCTCAA 240SUBSTITUTE SHEET (RULE 26)?W0 98/13501TTACCACTCGGCGGGCCCAAGTCAATTGATTCCAGAGTGAATGAGGCGGAGATGGTTCGATGATTCTGGGCAGACACGGCTAGTTGGTGAAGGACCTCTCGGAACAAACCGATTAGCCAGGACTGCATGAAAATGGGGGAGTGCAGGATCACTCCATGGGGGCAAGAGATGTATCACTGCGGATCAGTAGATGAAAATGAGAGAAGCCAGCCCATCCTCCCGCAGGACAGCGGAAGAACAACTAGGTGCAAAAACTTAGGATCTAGACTTACTAACAGGGTCAGAGGATCCCAGTCACAACCAATATTTTGAACAAGGAATACCATCCTAAGCTGATTCGATTTAGATTGCTTACGCCGACAGGATTGCTTTTTATCCTGATTTGCTGGTAACTGCACCAATACCCTCTGAGGTTTGAACGGTGAGGAGGCGAAGATGCAAGCGGTTGGAGCTACCGAGAAGAGAGCTACAACCGACACATCGAAGGTCAAGGCTCAGACAGAGGCCGAGAACCAGGTCCCACTGGACCGGTGCACTAATAGACCGATGACCTCTGGCCTTATCACATGATGATCTCAGATAGCTCAAATTTGAGCTAAGAAGAGAGAAAATTTCATGGTCGGAAATGATATACTATTAAGTGAGTTATCCATACATGGTAACTCTGGAGCTTTTGGCCGATTCAGCTGGAAAGGCTTGTTTCCCAGACAATTGGGGGGGTAAGAGAAACCACCCTTAGACAGCTGACGCCCACGGACACCCGACCAGAACAACACAGCCGC-106-TGGTCAGGTTGTATATTATCCTGACGTTAGCCTTCGCATCATCCAAGTAGTTGAAGTGCAGGGTCTTUCTGGTGGATAAAGGTTGGATGTCTCTAATCCTGTGACATTGATTGGGATAGACACTTGAGTCTCCTGGAGAAATGCCATGGGCTTACTTCGAAGGTCAGTTCCAGAGATCGCCCCAAGTGTCAGGAAGATATGGCCCAGCAGTTGACACTGCTGCTCAGGCTCTAGAGTGTAACATCCGCCTCAGCCCACCA02265554 1999-03-16AATTGGAAACCTTATTTGTGCATAAGGCTGAAGAGGTACCTAGTGATCAAAGACCCTGAGCGCAAAGGCGGTACACCCAAGGTGAGGAACGGATATCAAGTACATATATCAACTATGTATCTTGATGAATCTCAATTCAGAGTAGGAGTGTCCAGCATATCACATTGGCAAATGCATACTATTCCTACACGAGGGTCAAAAGCAAGTGACATCGGAGTCCGCAAGCCATGCAATGACAGAACCCTCCATCACCATCCACTSUBSTITUTE SHEET (RULE 26)PCT/US97/16718CCGGATGTGA 300GAGTCTCCAG 360TTAGAGGTTG 420AACATGGAGG 480TCCAGGTTCG 540GGATTCAACA 600GTTACGGCCC 660CAAAGAAGGG 720AGGATTGCCG 780AGAACACCCG B40GTAGAGGCAG 900CCTGCTCTTG 960CTTTACCAGC 1020AACAAGTTCA 1080GAACTTGAAA 1140TTCAGACTAG 1200TCTGAACTCG 1260ACAGAGGACA 1320GGTGATCAAA 1380CAGAGTCGGG 1440GCGAGAGCTA 1500AGCCAAGATC 1560GCAGGAATCT 1620GATCTTCTAG 1680ATTGTTATAA 1740CCCACGATTG 1800?W0 98/ 13501GGGCCGATGGCTCAAGGCCGATATCAGACAGGTCTCAGCACGCGGTCAGGAATCTCCAGGGCGGTTAAGGAGCACCCTCTGATACCGAGGGCTTCTGATGAGAGGCAACAGGTAGGGCCATTTGGAACGGCCCTCGGAACGCCGTACTGAAATAATGAAGAAAACAGCCTCTGCTGTTATAGCATATCCAAAGGATCCCAGGCAGAGATTATCCAAGGAACTAAAGCCGAGCATCACGCACGTTACCTGACAGATGCTGACAGAAGAGCAAGCCCATCGGACCCAGGACAAACCATGCCTGATCTGGAGACATCAAGCACGAATCCAAGACAGGAGGAGAGATATGCTATTTGAAACTGCACTTTCCAAAGCACTTCCGAAGATCGCGTCCATCAGGGCCTACAGGAGTGAAGGGGGAGATGGCCAAAATTGAAGGGGGACCTTGGAAGGACGACCCCACCAGGCCGAGCTGACAAATGGTCGGGAAAAAGTGTAATCCGTGACTCTCCTTGAAGATAATCAGGCACGCCATCTCACTGGCCGGAGCGAGCCCTCAGCAATTGAGCGATGACTGGGTTACAGTGCTGACTCTCAATGAATCTCACTGACCGGAGAAGGAGGGGCTTAGGAAAGACACCCATTTTTATTGACAAGGTGCACCTGACACCCGAATTATTATGATACACGAGGATAGTTGAGTCAACACCTCTCATGCAGATGTCACTGGCTGAAACGGACCAGTGATGAGCTCACTCCATTATATGATGACATCAATGAAGTAG-107-GTCAAAKACGATCGAGGAAGGCCTGCAAGGGGATCAACTGGACGCTGAAATGTTATTATGATCATGGTTCGAAAACAGCGGGATCTGCTCGAGATCCACGACTCTCAATGAAAAAGGGCAGGTGGTGCAAGCGGGGAATGTCTGGTACCAGATGAGCTGTAATCAGAAGAATCAAGAAGCAGCATCATGAGAAATCAATCGTTCTCAAGATCCAGAGGACGCCGTCGGGTAAATCCAGCCAAAGGAGCCACTACAGCTCA02265554 1999-03-16GACTGGAATGCTATGGCAGCAAGAGAAGGCAAGGCGGTGCCTTTGGGAATTTTATGATCAAATCAGGCCTATGTGGATATCCATCTCTATAGCTCCTGAGTTCCCCCGCCCAGACGCGAGCCCAATGTGCTCCCCGAGTGCAATCTCCCCTCTCTGATGTTAATCACCAAAGATCAACAGTCGCCATTCCCCGACTTGAAAACCCGTTGCAGCTGCTGAATTGTTCCGGAGGCTAGAGGAACGATCTTGCACTTACCTGCSUBSTITUTE SHEET (RULE 26)PCTIUS97/16718CATCCGGGCT 1860ATGGTCAGAA 1920AAGCAGTCCG 1980ACCTCGCATC 2040CCCCTCAGGA 2100CAGCGGTGAA 2160TGATGGTGAT 2220TGGCGAACCT 2280GGGGTTCAGG 2340ACTCCAATCC 2400CCCGGACCCT 2460ATTAGCCTCA 2520TCGAAAGTCA 2580TGTGAGCAAT 2640GAGATCCCAG 2700CCAAGATATT 2760GCTAGAATCA 2820GCAAAATATC 2880TGGACTTGGG 2940ACCCATCATA 3000CAGCCGACAA 3060GGAATTTCAG 3120CACCGGCCCT 3180GGATCGGAAG 3240CAAGTTCCAC 3300CAACCCCATG 3360?WO 98/13501CCAGTCGACCGCCTCCCAAGAAGGGTCGATTCAGAGTCATTGCTGGGGGTCTCTGCCCTTCTGAGCTTGAACAACACCCCTCAACGCAAATCCGTGTTGTGAAGAATGCTGGATTGACAACAACATTTATATTATTGCAAGCACCAGTCTGGTTCAAGAATCTGGAGGAGAAGAATTCCGTGTAGACCGTGCCCGGAAAAGCTGACGGCACCACCAGCCATGCCCCCCACACTGGAAGAGGACCGAGGTGACTAAACAAATAGCTAATACTTCCACAATGCGCTCCGATAAGATCCTGGTTGTTGAGGACAGGTGTTGGCCATAGTTGTTACTAACTCTCCCAAGTGTGCTTATATGAGCGGAATTCAGAGGCGATTGGCGGTCCACATCAATGAAAATCTCACATTAGAGACCCTATGTCAGATGCAAGCATTTACGACAGTGCCCAGCAAAGGCCCCCAGCACGAACATCCCAATCTGCCAAACCACCCCCTTCCCCTACCCAACCGCACTTAGGGCCCAAACCTAAATCACAGAGATCTCAACCCACCACTAGGCGACAAGCGATCTCCAGATCCACAGAGACGTACAGCTCATACCTTAATGCGGTTAATCACCCGTCTCGGTCAATGCATGGGAAGAGGGAACTTCAGAAAAGATGGAGCACAGGCATACCCACTGAATAGTAAGAAGACGTTATCAAATGCCCGAATCCGAAAGACCCAGGCGGCCCATCCTCCTCAACCGCATCCTTCCCTCAACAGGCACCCGAAAGGAACATA-108-CATTATAAAAACGACTTCGACCTACAGTGAGAAAAGATGATAGGGCCTCCCAAAACCCGACAGGGCTCAAGGAGAAAGGTATCTGATACCTTTCAGATAACAGTGGCCTTTCATCGACAAGGAGAAAGAAGCCTGGTTTTAAATGAGCAATGGATATCAATCCAGGCAGTTAAATGATGAGACGACCCTCTCCACAGACCCCAGCACAGAGTAGGACCCCCTACCACCCCACAAGAACTCCTCCCTAGACCACACCCAAC02265554 1999-03-16AACTTAGGAGCAAGTCGGCATGGCAGGCTGATGTTTTATGAATCGGGCGAAGAACTCCTCTGAAAAACTGCCTAACAACAGCTGGATACCCGGGTATTACCAACCTGCTGTGCAGAGCAAAAGTGAAGTCTGCACTTGGTGACTCTCCATTGAAGACCTTTTTGCAGCCACCAAGGATTACTCACAATGAAAATGAGAGGACAGCCCTGACGAGGACCAACGGGAAAGAACACAACCGAAAGATCCTCTCAGAACCCAGASUBSTITUTE SHEET (RULE 26)PCT/US97/16718CAAAGTGATT 3420TGGGACATCA 3480GTGCCCCAGG 3540TACATGTTTC 3600GCATTTGGGT 3660AAAGAGGCCA 3720GTGTTCTACA 3780GGGAGTGTCT 3840CCGCAGAGGT 3900ACCGTTCCTA 3960GTGACCCTTA 4020CTTCCTGAGG 4080TACTCTGCCG 4140GGGATAGGGG 4200GCACAACTCG 4260AATCGATTAC 4320TCAGTTCCTC 4380TTCAAAGTTC 4440CAGCCAGAAG 4500CCAGCCAGCA 4560CATAAGGCCA 4620CCCCCAAGGT 4680ACCCCCAGCA 4740CCACACAAGC 4800CCCCTGGCAA 4860CCCCGGCCCA 4920?W0 98/13501CGGCGCCGCGCCCCGGTGCCAATCCAAGACGAGGAAGCCCGGGCCACCAGACCCCAGCCCCGAAGGACCCCTCTTCCTCTCCCGTCCCTAGGTCTCAAGGACCGGTCAAAAGCTACAAAGATAACTCTCCACAGTTTTGGCAGAGTGTAGGCCCTAGGCGCTGAACTCTCGAGGCAATCAATCAATAATGCTCGGGCTCACGGGACCCCAATCAATAAGGAGCAGAGGAAAGTATAGCCTGTCTCGTACACAAGGGTACCCCCCCAACCCCACAGGCAGGGGGGGGGCCCACCCACCCCATTCCCAGACTCGATCCGGCGCCGAACCGCATCTCGAAGGGAAGGAGACACTGAACGTCTCTCCATTGGGGTTATGACTCGTCAATAACTGAACCAATTAGCTTCAAGTAGTTGCCACAGCAAGCCATCGAGACAAGCAGGAGCTGATACCAATTGCTCAGTATCTGCGGATGTTAGAAAATAAAGGCCCGACCCGACGCTACATAGGCTCTTATCTCGAACA 02265554 1999-03-16CCGACAACCACACACCAACCCCCCAAAAAACACACGACCACGGCCATCACGGCAGCCACCAAGGACATCAACTAAAAGATCGGGAATCCCTGCCATATTCCAATCTCTCTTTCCAGCCATCACGAGGGTAAGATGCACTTGAGACACAAGTGCTCAGATACAATCTGAGGGCAGGAGATGGTCTATGAACATACTATACAGATATCTATCGCTCGGATATGATAACTCACGTCCGAGATCTCAAGAGTGGTTTTGATGAG-109-GAGGGAGCCCCCCGAACAGAAGGCCCCCAGCGACAACCAACCCGCAGAAACAACCCTAACGTATCCCACACAATCCACCAGGAATTAAGAATGGCAGTACAAGATAGGGGCAATCATTAGGAGATTGCAGAATGCAATGAAGATTTGCAGACAGCCGGCAGCAAGTCTGGATATTGGCTGCAACTATCTTGAAATCCTGTCAGGCTTTGAAGTGGAGGTGGTCGACACAGAAGGGGGTGATATACGACTGTCATCGTGTACCAACCAATCCCCAGCACCCGGGCCGACAGACCAGAACCCGGAAAGGCCACAGCACCCAAGCCTCTCCAACATCCGACGACTCATCCAATTGTTAACTCTTGGTAGGAATTCATAAAATTAATACAGGAGCCCAGAATATGAGTAGTCCTTTGCACTTCAAAACTACTAATTCAGGGTGTGTGATTTAATCATTATTTGGGCTATGCGCTATTTACTGGGAGTCCTACTTTTGTCCACCGTGCCCAAGTACTTTCATGCCSUBSTITUTE SHEET (RULE 26)PCT/U S97! 16718CCGCCGGCTC 4980AGCCATCGAC 5040CCAGCACCGC 5100AGACCACCCT 5160CAACCTGCGC 5220GAGCGATCCC 5280GTCCCCCGGT 5340CACTCAACTC 5400GTCCATCATG 5460CCAAACACCC 5520AGGAAGTGCA 5580AATGCCCAAT 5640ACTACTGAGA 5700AAGACCGTTT 5760GGCAGGTGCG 5820CCAGTCCATG 5880TCAGGCAATT 5940CCAAGACTAC 6000CGGCCAGAAG 6060CCCTAGCTTA 6120CGGAGGAGAT 6180CATCTTAGAG 6240CATTGTCCTC 6300GCTAGAGGGG 6360TGTTGCAACC 6420AGAGGGGACT 6480?W0 98/ 13501GTGTGCAGCCTCCACCAAGTTCACAAGGGAACGATCATTAGTAGTCGAGGTACTTGCACAAATCTGGGGACAGATATTGAGTGTGTCTTGAACAAAAAGGACATCGAAATCACAAGTCTCTATCTCCGGCATCATCCACATCCCAAGGGATTTGCTGGCTCATTAGACTTTCTAGATGTAAATCATCGGTCATCTCTGACTTGGTGTATCGGCTGCTGAACAATCAGTTCATTCTCAAACATCTATAGTCTAATCTGAGCAAAATGCCTTCCTGTGCTCGACCTAATAGCATCAAGACCCTGAACGGCGTGAATTGACCTATGCAATTGCGGAGTATGAAGAGGGTTGATGAGAACAAGTCCTATGTAAGCTCTTCGTCATCCCCTTTGGATGTCACCACAGTAGGATAGGTTCTGTTCGCATCGGGCAGACTAACTCAAGATGAAGTGGAAGATTAAATAACCCGCCAGGAGCTCATGACTAGCTGTCTATGTCGCTGTACTATGACATAGCAAAGGGTCAGTACCCGATGTACACTCGTACAATTGTGCATGACAAGATCGACCATCCAACGGTCCTCCCTAAGTTGGAGAGGTTTGTCGAGGGATCCCCTGGTATGTCAGTCGCTCTGATCAAGCAACCCCGAACAATAAACGAGACCGTTATCAACAGTCATGTTTCTCCATCTACACTTGAGCATCAGCCTGAGGACTCCTTAACCCAGAGAATCAAATGCATTGGTCAAAGGGAAACCCTGTTGGACCCAGGGAATCAGAGTTGTC-110-AGTCCTCTGCTCTGGGTCTTTCAATCCTTTCTAACATACAGTCGGGAGCAATATCATTGGGATGCCAAGGAGCACTAGCAGCTTTAATATAGACCAGGCCTCCTCTACAAACCGCATCCATCGGTAGTTAGATAAATGCCAGAACACCTTGAGCTTGATCCGCAGAGATCGGTCAAGGACACCTCAGAGAGGATAGGGAGATTGGATTATGAACTCAACTCTGCTCAGGGCTTGTATTTAGTACGGGGGAACAACTGAGC02265554 1999-03-16TCCAAGAATGTTGGGAACCGGCAAGTGTTATTGCTGCCGAGGAGGTATCCAGAGGTTGGAAATTGTTGGATAGTCTACATGTTGCTGCAGTAAAGCCTGACTCTTGGAACGCATCAAGCCATTAAAACTTTTCTACAAAGATGATTGATAGGGTTGCTAGCATAAAAGCCGTGCTGACACTTCACTGACCTACGACTTCAGATCAATACTCTACTGGAGACCCACTACAAAGTCGAGGTTACTTACCTAGATGTACCGAGSUBSTITUTE SHEET (RULE 26)PCT/US97/ 16718CCTCCGGGGG 6540GTTCATTTTG 6600CACAACAGGA 6660TCACTGCCCG 6720AGACGCTGTG 6780CGTAGGGACA 6840GTCATCGGAC 6900CCTGATTGCA 6960GGGGCGTTGT 7020TCTTACAGGA 7080ACAAATGTCC 7140CACCTGAAAT 7200AGGGTGCAAG 7260ATAACCCCCA 7320GACCTTATGT 7380CAATTGCAGG 7440TCAGCACCAA 7500CACTCTTCAA 7560TAGTGAAATT 7620GAGATCTCAC 7680GTGCAGATGT 7740CCAGAACAAC 7800TCAGAGGTCA 7860ACAATGTGTC 7920TGGAAAAGCC 7980TGTTTGAAGT 8040?WO 98113501AGGTGTTATCGCAACCAGTCAGCCCTTTGTCAGCTTTCAGCCCCTTCTCATATCGCTGACAATGGAGACACGAGTGGGCAGAGTCTAACACGGTTCAGGGGCCAATGAAGGGTTAGTCCCAACATACCTAACCTGGTCAATGTGGTTTATGCCTATAAAGCTGGTGCCGTTGGGATGGTGATAGGGCTGCGTGAAATAGACGCTATCTGTACAAGATAGTCTACACTGTGTAAACAATGTCTCATATTCCCGAGGAAGATAGAAATCCGGAGTAATGATCCACGGGGGAGCTCGTCAAGCACGGATGACCAATCAAGCAATGCTTCCAGCCCATTGAAGGGTTGAGCTTAATGGACCTATAACCTAGCCCAACCTCTTCACCTGCGGAGGGATCTCCAATTACGTTTACAGGGATCCCCACACTTCTGTGGGCATGGGAGCAGTGAACCACATCAGAATTCAACCAGATCAGCCATCCTGTCAGAACATCGGAAGTTGGGATATCCAAACCCGTGAACTCCAGTTTGGGGGCTCAGCAACTGATTCTATCACTAGGTGTCTGCAGTGATAGAAATGGGCTATAGGCGTGTAAATAACAGGATAAATCAAAATACAAGTCCAATAGGTGTAATCTGTCCCAATTGGATGGTGAATGTTTTGGCGCCCAAGCCGTCGAATTACATGCTTGCGGATCAGCTGCACATCACATGATAAGAAAAACGTTATACCCCGGAGTATGCTCAAGCACCGCCAATGTCATCATGTAATCAGGCTCAAAAAGG-111-TCCGGTGTTCTATGGTGGCTAATTCCCTATGAAATCOCCACAGGCTTTACCCCGACAACAGGGTAAAATCTCCTTCATACTGCTTCGGGACCACAACAATCAACACATTGTAAGGAAGCATGTCAAACTCAACCTACGATCTCATTTTCTAGTGGAATGCCTCAGAATCTAGTCACCCGGGTCACCCAGATAGGGTCCAAAAGTTCACCTGAGTCCCTCATAAAAAACGGAGTCCAAGCTATTTATTTEAGAAATTCGCT02265554 1999-03-16CATATGACAATTGGGGGAGCCAGGGATCAGACCGACATGCCTCTCATCTCAGAACAGATGCAAGCACTCTGGAGTCTTGTTTCGGGCCATGAGTATTGGCGAGTGGATACGGCGAAGACTAGTTCCAATCACTTCCAGGGTACTTTTATCTTCACATGGGGGTGGACATAGAAGATGGAACATCAGGCATGTGGTTCCCCAGATAGCCCGCGCTTACAGCATTTTCCAACTAGGAGTTATCATAGAAGACGTACTCTAAASUBSTITUTE SHEET (RULE 26)PCT/US97/16718ACTATTTTGA 8100TCAAACTCGC 8160GGAAAGGTGT 8220AATCCTGGGT 8280ACAGAGGTGT 8340ACAAGTTGCG 8400GCGAGAATCC 8460CTGTTGATCT 8520TGATCACACA 8580TGACTATCCC 8640CGAGATTCAA 8700GCCATGCCCC 8760TGGTGATCCT 8820TTGAACATGC 8880CTTTTAGGTT 8940ACCAAAAACT 9000TCACTCACTC 9060CCAATAGCAG 9120ACCCACTAGT 9180GTTATGGACT 9240ATAGTTACCA 9300CTGGAGGACC 9360CAAATGATTA 9420CCGGCCCACT 9480AAAGAGTCAA 9540GTCAGTAATA 9600?W0 98/13501AGGTTTTCCAAGGACATCAAAGCCCTTTCTCCCATACTTGTGCTAATCTCTAACATTTGACTGCTATGACAATTGATAGATGGAGCCTCTCTTTCCTTAAATGAAGGTACTACACCTGACCAGTAACGGCAGACTCTGATGGCACGGAGGATGCTCAAGCTTGCTGGAGTACCTAAAGGAAGTTCCTGCGTTAATGATTCTCCATGACCCGTAGACTTTTTAATCTCAAAATTTGACTAAGTCACAGAGGGGAACGTGAGATGCTTGAGGGGAGAAAGTTGTTTTGGTTTCCATAGGAGGTCGTGACCTTGCTGGTTTTGCATTGATGCTTGGTTTCTTCTTCACTTGCTCCACTGCTTTTTATCACGAGAGGGGAGATTTGCTGAAAATGAAAGGTCATCAGTTGGCCACTCAGGTGAAGAAATTTGGCCAAGGCACTTTTACGACCCCGAGCTTTGACTGAGTTCAACTGCTAAAATGCGGGATTGGCGGCACTCCACGGGGCCAGTCGGCAGCAAAACAGACACTAATTATTAACTTGGACAGTCAAGAAGACACACACGTTGCTATAAATGTATTGTGAGATATACAGCCTGCACTCGTACCTGCAGCACTGAAATACTTAGTTGAAGTTCTCATTTTGTTAGGAAATGCCATATTTTCCGCTGACCCGGATTAACACTGCTTCATGCGCTGCTCTCCCCCAAGGGAACCATATGATACTGTCTTACAACTTACAAAAAAATATTTTAACTCTGGCTGCTAAAAACCTGGGTTTATAG-112-CACGGCTTGGGAGTTTACATCTGAGATGAGCTGTATTCTTTCAGTAAAGAATGTCATAGAAGCTTCTAGGGGAATCCAACTGAGGGATATATGATGTTCTCTCTAGATTATCAGAAGTTTACATGAATCAGTGGAATCATTCCCCCTGCAATGAGCAGTGCTCTTAGCCTAAAGGGAATGCCGGGTCACGTGATAATGTAGCCTGAAAGATGAGGGCATGAGGACAATGGTCTCAGGAGTACTCCCGAAGGGTTCCCTCA02265554 1999-03-16TCTAGGCTCCGCACAGCTCCGTCAGTGATTCACTGGTAGTGTCTCAACATGGGGAGGTTAAAGAGTCAGATTATCAAATTAACGGTAGAATGACCAAAACCATTTTCATACGGCCACCCCGCCTAAAGTCAATCAACGGCTGCTGCAGACCGTTGATAACGGATAGTGATGGATTCAGTTGAGGCTTGTATGTTGTAAGTAAAGGAGATCCCAAGTGATTGATGGCCAAGCCCTAAAGATCCCAGCCCACGATAATTCGGSUBSTITUTE SHEET (RULE 26)PCT/US97/1 6718GAATTGAGGG 9660CAATGGTTTG 9720AAATCACAAA 9780TCAGTTGAGT 9840GTATATTACC 9900ATGACAGAGA 9960TACATGTGGA 10020GTAGCCATGC 10080CTCAGAGGTG 10140GGGTTTTCTG 10200ACTGATGACA 10260AGACTTGAAG 10320ATTGTGTATG 10380TATCGTGACA 10440ACAATCCGGA 10500TGGAAATCTT 10560CTGACAATGT 10620TACCCGAAAG 10680GATGTTTTCC 10740GGAGCTTACC 10800AAGGAAACAG 10860GCTGAAAATC 10920GATGAGCACG 10980CTCAAAGAAA 11040ACAAATACCA 11100CAGGACCAAG 11160?W0 98/13501ACACTAATCAATCTCAAGAATAAATGAGATCTGTCCTGTAGCAAAGTCCCGTCAGAAGCTGAGTAAGGATTACCCAGCACACTTTGTAATCAATTGTTTCTGTCCCAATCAAACAAGGGCATGACCGTTACTCTTGGCTTACAACGACCTTGAATATGAGATCTCAAGAGCACAGCAACCTTGTATGTGTTCCACAGTCCAGGGACTGGCTCCTGGATCAAAGGCCTGATTGTCCAATTAGAAATGTCCTATATGTGGGCTCCGGAGAATGTACTGCCTTTTACGGATTATGTAAGTGACCAATGACCAAGTGGACCATCTGCTTCATTAATGGCCTTACTCTTAGGCAAATCACATTTTACTCAAGAGCAGCATGCAGT-CCTTGCATATCACAATCAATCTTAATAAGGCAGGCTGTTTAATGATTCTCGGGGGACTCTCCAGAGCATCAAACCCAATGAGCATTCCTCTAGTGTCACATCGAGCCAGCTGACTATGAACATTGACAAAGAGGCTAGCTCAATGGAAGCTTAATTGGAGATCCCTCATTTTCCTCATTGCCATCTTCAITAAGCACCATTCGTGCAAGGGGAACCTTAAGAAGGCTACATGTTTGTTTATTATCGCAAGATAATATTGCTATCCCTGAACGTCAACCATGAATGGCACTGTGTCAGAAACAGCATCACTGATCATTCCTAGACTAGACTCCTTAAAGGGATATGGACAGGCGGGGCAAGAGATGAGGAAGGCAATTCAGAGGAGTCATGTTCGAGGACGGC-113-ACGAGACAGTATGAGACCATTTCAGTGGCTCCCCCGACCTAGTACCCTATCCTATTTATAACAATCAGACAATGGGAAGCACATTGGCCACAAAAGGAATGTGTATTCTGCAACAATGGCTCCTAAAAGTCCCAGGATGTTGCCCGCTCCTCGGTGATCCTGCCTGAAGAACTGGGCTAGTCAAGAACATTATTCCATGAATATTATAGTAGTCTATTGCGGGGGTTAACCAGGGATGGTCAGTGCAGCTCTATTTACGG02265554 1999-03-16CAGTGCATTTCAGCTTGTTTGCATAAGAGGTGACGCCCATGGGAGGTATACCTGGCTGCTCATAGCTGTATGCTAGAGTATCACCTCAAGATATTATGATGTCAGAGACTTAAAAGCATCGATACAGCAGAGTCATACCCTATTGGGGGGAGTAACATCAGACCCTCCATCGACCCTTACAACTGCAAGGTGACAGTAAAACCTAGGGCAAGGCATGCTACTCTCGAGTGGCTATTAACAGGCGAGAGCCCCTTGAGGTCSUBSTITUTE SHEET (RULE 26)PCT/US97/16718ATCACAACTG 11220GCACAGAGGC 11280CTTGAGACCT 11340ATCCCGTTAT 11400GAAGGGTATT 11460TATGAGAGCG 11520ACAAAAAGGG 11580ACTAGAGATT 11640GCAAATGAGA 11700GGGCTACTTG 11760ATAGTTGATG 11820GAGAGAGGTT 11880ATTCTGATCT 11940CTCCTCACAA 12000ATGAATTATC 12060TCAATTGCTG 12120CAAGTAATGA 12180TCAGCAAATC 12240TTTGTCCTAA 12300GAAGAGGACG 12360GCTCATGAAA 12420GATACCACAA 12480ATAACCAGAT 12540GGAAGAAAGA 12600CTAAGAAGCC 12660CCTGATGTAC 12720?WO 98/13501TAGAATCTATGATCAGTCAAAGGAAACATCTGAAGCTTGCCAGTGTACTCCAAGGCAAAGCGACTAATTTCCCTTGTCCGCAGATAAGAATTTTAGAAACTTCACGTCGACTCGCAAGCTCTTTAATTGAAATTTGTTACCTATGATTGATAGGGGATGATCACTATCTAGACCATCAGGAAGGAGTGTTGGCACTGTGGCAACTGTGTGAAGAGTTAGAGATTCGACAAGGACCTGCCCATATCAAGGCTTGTAGACCAGCGAGGCCACCTACGGATGGATCCTTGAGACTTCGTAAGAATGGGCTTACGGCTAATGTGAGCACATAGGAGTGGCAAGGGGTTGATACTATTGTTTCGAAACAGATTGTAGAGCTGAGGCAGAGATGCAATGGTCCACACCTGGTAACACGATATCAATCTTGGGCCAGGAAATATCAGTAAGGTGCTTTATTATAGAGCAACATGGTTAGAGTTTACACATCCAGGCAACCAATTCGAGGAGGCTAGGTTACTCATGCCACTTATTCGGCTTTTTTGTCCGTCCCATATAGCCCCAAGTCGGTGATGATGAGCCTGGAGGTTGAGGGATCTATACCACAAAACTTTATATCTCGAGAAAGTGCGTGATCCGCAGAGCTGTACAAGGCTATCCCCAACTATAAATTTGAGAAGTTTCATAATGTGCGGCCAATGGGTGAGCGTCAATGCTCCCTATCCATGTACACATGCTTTTCTTTTGTAAACACTTGTGGTCTAAGACTTATCTCCAGTCTCTGACTT-114-GTCATGAGACCCTCGGGTTGTTGGTTCTACGATCCTTGCGATAGCTCTTGAGCTAAGGGTGTAGCACTCATCTCCAACGAACCAACAAGGATACCGGATCCAATGATAGAGTACCAACCCACACCCAGAGATCACATTCTAGGACCATATCTGAGTTTCTTCAATTGGGCTGTTGTCATCTAAGCCACCCGTCCTTCACTATATGACCTAGTGAAAGTGAGTGTTCTGGCCGGTAGAGAACAGGATCTTCATCTTCGGCG02265554 1999-03-16ATGTGTCATCCCAACTGGATCACTGATGAGATCTGCTGTTGAACGAAGCCGATCACTCCCAGTGAAATACCAATCTCTCAAATGCTCCTAATCTAACACGTCATCCCAGGATTGATATATCCATAGGAGGAGCTAAGTCCGAATGAAATTGCTTATAGAGATTTGATGTAGTTCCTTTCTAAAGATCTACTGATGCTCAACCTCGACCTGCGAGGATGTAAGATTTGTACATGTGCAGTTGTGGAACATAAGGATCGATCSUBSTITUTE SHEET (RULE 26)PCT/US97/16718TGCGAGTGTG 12780GATATTGACA 12840AGAACAGACA 12900AGAATAGCAA 12960TGGTTGTTGG 13020ATCTCAACTT 13080TCAGGTACAT 13140TTTGTCATAT 13200GGGTTGGGCG 13260GTATTACATC 13320ATACCCAGCT 13380GATAATGCAC 13440CACCTTGTAG 13500ACAGCACTAT 13560TCAGCTCTCA 13620CCAAGATTAT 13680CATTATCATA 13740AGAATGAGCA 13800AAGAAATTCT 13860AACTTGCACA 13920TTGTTGAATG 13980GTACCGGACA 14040TGTCAACCAG 14100CTAACCGACC 14160AATCCAATTA 14220AAACAGATAA 14280?WO 98/13501GATTGAGAGTCAAAGATCGGATGATGTTGCGGGGCAATCTCTTGCTACAAACGGCTTATTAACTAAACAAAATTAGCACCTTGTCAAGGTTCAATTACATAGACCTTACCTGGCTCTGCTGGGATTTTGTTATACCCCAGAGGCTAACCGGGACTTCACCCAATTGTGGGCTATAGAGCAAATTGATCCATCTACAGGGACTTACCCCGTTTTGGGGGCAATCTCAAGTCCCAAGTCAGATAACAGTCAAACTAATTGGTTGATCCAGGACAGCAACAACAAAATTGCTCCGCCAATTATAGCTGTTGAGCTTGGGTGAGGTGCTTCTATCTATCCCTCCGCTCTTTAACAGTTAGTAATTAACAAAGATCCTGGGCAAATCAGGGATTTATACAGCAACGCTAATGAATTGGACTTATAAGACGCAGTTGGTGCTGATCCCATGATGTTGTTGGCAAGAATTGGTAAGTCATTCTTCTTCGGTTATCTGGAAACAGATTGGAGACCAAGTGAACTCCGGCATTCATTTTCGATCTCAAATAAAAGATATCAGAAATCCATGATATCAACATGGATCGGGTTAATAGTGGGGGAAGTTGGCCGGGAGGCCCGATCCCTACCTACTATAGAGAATAGGATCAAATAAGTTATGTTCATATCTACCTGAAAAGAGGTCACATCCAGTAGAGGTGAATTGCGGGTGCCTCAGGGCTTCAAGGACAAGCAGGCAACTACTCCGGGAATACTAGACTATTATGACGGGAATGGTATAAACCCTAATC-115-ACGCCCTCGCTGAGCATCAAATACAAGCAACTTTCCGCAGTAATTAGGAGCTATGTTGATTCTCTGCCAATTGTCGAACAAAGTCACATGCTAGTGTGGGAGCTAGAGGATACTGGTGATATAGGGTCTCACTGAATCTTATTAAGCAGCATATCCATTAAATATCAAICCTGGCAATTAAAAGATGGATTACCAAAGAAGGAGAACTTATACAGAAAGTTTACACCAGAAGGGGTTTGAAAGTTAGTCGGCTGCCCCAGG02265554 1999-03-16TGAGGTAAATGGATTTCAGAGCACAATCTTAATCGGGTTGATGCCTTGAGCACTTATAAGTTCTAGATCTCAGAATGGGAGGTAGGCAGTGTTTATCCATATTGGCAGCCTAAGCTTATGTTATAGAGAATTTGGTTATGGATAATTGAAGCAACTAAGCTACTCTGAAACGGACCTAAAGCTTAATTCTTCAACAAGGGATCTAGAATCGATAAATAAGTATCTTCGTTACGTGAGTGGATACAGTGCCTGGTTAGGCASUBSTITUTE SHEET (RULE 25)PCT/US97/ 16718GTCAGTCAGC 14340CCCCCACACG 14400CCCATTTCTG 14460AACTCATCTG 14520CCAGGGGAAG 14580GAGATACTTA 14640GGTCAAAGGG 14700GTAGGTAATA 14760GTAGATTGCT 14820TCAGATATAG 14880ATCTTATCGA 14940CCTTTCAGCG 15000GTGAACCTTG 15060ACAGATCTCA 15120TCATCTGTGC 15180TGCATACAAG 15240AAACTTACAC 15300CTGTGCAAAG 15360ATACTCATCC 15420ATGTTCCACG 15480ACTCGCAAAT 15540TTTATCCAGA 15600AAGAATCTAT 15660GTTTTTAAGG 15720CTGATTAAGG 15780TTATTTGTAA 15840?W0 98/13501CA- 11602265554 1999-03-16PCT/US97ll6718TATATTAAAG AAAACTTTGA AAATACGAAG TTTCTATTCC CAGCTTTGTC TGGT(2) INFORMATION FOR SEQ ID NO:4:(i)(ii)(xi)MetAspArgIleAsn65AlaIleGlyArgIle145TrpSerSEQUENCE CHARACTERISTICS:LENGTH: 2183 amino acidsTYPE:STRANDEDNESS:(A)(B)(C)(D)TOPOLOGY:amino acidlinearMOLECULE TYPE: proteinSEQUENCE DESCRIPTION: SEQ ID NO:4:Asp Ser LeuSerVa1Lys50ValHisGluAsnAsp130LysPheValProPro35HisGluSerAspSer115ThrGluGluIleIle20HisArgValHisLys100LeuAsnLysProLys180Ser Val Asn Gln Ile5Val Thr AsnAla SerTyrAsn55Leu LysAsn Val70GlyIle85ProTy:Glu Ser ThrTyr Ser LysLeu135Ser ArgVal Ile150A611Phe Phe165LeuSer Gln ThrSUBSTITUTE SHEET (RULE 26)LysLeu40GlyIleProArgVal120GlyLeuTrpHisIle25GluPheLysAsnLys105SerLeuGlyPheThr185Leu10ValAspSerSerCys90IleAsnGlyValThr170CysTyrAlaProAsnLys75AsnArgLysSerTyr155ValHisProIleThrGln60LeuGlnGluValGlu140MetLysArgGluLeuLeu45MetArgAspLeuPhe125LeuHisThrArgValGlu30CysIleSerLeuLeu110GlnArgSerGlu’Arg190His15TyrGlnIleTyrPhe95LysCysGluSerMet175HisLeuAlaAsnAsnPro80AsnLysLeuAspGln160ArgThr15894?CA 02265554 1999-03-16WO 98113501 PCT/US97l16718-117-Pro Val Phe Phe Thr Gly Ser Ser Val Glu Leu Leu Ile Ser Arg Asp195 200 205Leu Val Ala Ile Ile Ser Lys Glu Ser Gln His Val Tyr Tyr Leu Thr210 215 220Phe Glu Leu Val Leu Met Tyr Cys Asp Val Ile Glu Gly Arg Leu Met225 230 235 240Thr Glu Thr Ala Met Thr Ile Asp Ala Arg Tyr Thr Glu Leu Leu Gly245 .250 255Arg Val Arg Tyr Met Trp Lys Leu Ila Asp Gly Phe Phe Pro Ala Leu260 265 270Gly Asn Pro Thr Tyr Gln Ile Val Ala Met Leu Glu Pro Leu Ser Leu275 280 285Ala Tyr Leu Gln Leu Arg Asp Ile Thr Val Glu Leu Arg Gly Ala Phe290 295 300Leu Asn His Cys Phe Thr Glu Ile His Asp Val Leu Asp Gln Asn Gly305 310 315 320Phe Ser Asp Glu Gly Thr Tyr His Glu Leu Val Glu Ala Leu Asp Tyr325 330 335Ile Phe Ile Thr Asp Asp Ile His Leu Thr Gly Glu Ile Phe Ser Phe340 345 350Phe Arg Ser Phe Gly His Pro Arg Leu Glu Ala Val Thr Ala Ala Glu355 360 365Asn Val Arg Lys Tyr Met Asn Gln Pro Lys Val Ile Val Tyr Glu Thr370 375 380Leu Met Lys Gly His Ala Ile Phe Cys Gly Ile Ile Ile Asn Gly Tyr385 390 395 400Arg Asp Arg His Gly Gly Ser Trp Pro Pro Leu Thr Leu Pro Leu His405 410 415Ala Ala Asp Thr Ile Arg Asn Ala Gln Ala Ser Gly Glu Gly Leu Thr420 425 430His Glu Gln Cys Val Asp Asn Trp Lys Ser Phe Ala Gly Val Lys Phe435 440 445Gly Cys Phe Met Pro Leu Ser Leu Asp Ser Asp Leu Thr Met Tyr Leu450 455 460SUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/ 13501 PCT/U S97/ 16718-118-Lys Asp Lys Ala Leu Ala Ala Leu Gln Arg Glu Trp Asp Ser Val Tyr465 470 475 480Pro Lys Glu Phe Leu Arg Tyr Asp Pro Pro Lys Gly Thr Gly Ser Arg485 490 495Arg Leu Val Asp Val Phe Leu Asn Asp Ser Ser Phe Asp Pro Tyr Asp500 505 510Met Ile Met Tyr Val Val Ser Gly Ala Tyr Leu His Asp Pro Glu Phe515 520 525Asn Leu Ser Tyr Ser Leu Lys Glu Lys Glu Ile Lys Glu Thr Gly Arg530 535 540Leu Phe Ala Lys Met Thr Tyr Lys Met Arg Ala Cys Gln Val Ile Ala545 550 555 560Glu Asn Leu Ile Ser Asn Gly Ile Gly Lys Tyr Phe Lye Asp Asn Gly565 570 575Met Ala Lys Asp Glu His Asp Leu Thr Lys Ala Leu His Thr Leu Ala580 585 590Val Ser Gly Val Pro Lys Asp Leu Lys Glu Ser His Arg Gly Gly Pro595 600 605Val Leu Lys Thr Tyr Ser Arg Ser Pro Ala His Thr Asn Thr Arg Asn610 615 620Val Arg Ala Ala Lys Gly Phe Ile Gly Phe Pro Gln Ile Ile Arg Gln625 630 635 640Asp Gln Asp Thr Asn His Pro Glu Asn Met Glu Ala Tyr Glu Thr Val645 _ 650 655Ser Ala Phe Ile Thr Thr Asp Leu Lys Lys Tyr Cys Leu Asn Trp Arg660 665 670Tyr Glu Thr Ile Ser Leu Phe Ala Gln Arg Leu Asn Glu Ile Tyr Gly675 680 685Leu Pro Ser Phe Phe Gln Trp Leu His Lys Arg Leu Glu Thr Ser Val690 695 700Leu Tyr Val Ser Asp Pro His Cys Pro Pro Asp Leu Asp Ala His Ile705 710 715 720Pro Leu Cys Lys Val Pro Asn Asp Gln Ile Phe Ile Lys Tyr Pro Met725 730 735Gly Gly Ile Glu Gly Tyr Cys Gln Lys Leu Trp Thr Ile Ser Thr IleSUBSTITUTE SHEET (RULE 25)?CA 02265554 1999-03-16W0 98l1350l PCT/U S97! 16718— 119 -740 745 750Pro Tyr Leu Tyr Leu Ala Ala Tyr Glu Ser Gly Val Arg Ile Ala Ser755 760 765Leu Val Gln Gly Asp Asn Gln Thr Ile Ala Val Thr Lys Arg Val Pro770 775 780Ser Thr Trp Pro Tyr Asn Leu Lys Lys Trp Glu Ala Ala Arg Val Thr785 790 795 800Arg Asp Tyr Phe Val Ile Leu Arg Gln Arg Leu His Asp Ile Gly His805 810 815His Leu Lys Ala Asn Glu Thr Ile Val Ser Ser His Phe Phe Val Tyr820 _ 825 830Ser Lys Gly Ile Tyr Tyr Asp Gly Leu Leu Val Ser Gln Ser Leu Lys835 840 845Ser Ile Ala Arg Cys Val Phe Trp Ser Glu Thr Ile Val Asp Glu Thr850 855 860Arg Ala Ala Cys Ser Asn Ile Ala Thr Thr Met Ala Lys Ser Ile Glu865 870 875 880Arg Gly Tyr Asp Arg Tyr Leu Ala Tyr Ser Leu Asn Val Leu Lys Val885 890 895Ile Gln Gln Ile Leu Ile Ser Leu Gly Phe Thr Ile Asn Ser Thr Met900 905 910Thr Gln Asp Val Val Ile Pro Leu Leu Thr Asn Asn Asp Leu Leu Ile915 920 925Arg Met Ala Leu Leu Pro Ala Pro Ile Gly Gly Met Asn Tyr Leu Asn930 935 940Met Ser Arg Leu Phe Val Arg Asn Ile Gly Asp Pro Val Thr Ser Ser945 950 955 960Ile Ala Asp Leu Lys Arg Met Ile Leu Ala Ser Leu Met Pro Glu Glu965 970 975Thr Leu His Gln Val Met Thr Gln Gln Pro Gly Asp Ser Ser Phe Leu980 985 990Asp Trp Ala Ser Asp Pro Tyr Ser Ala Asn Leu Val Cys Val Gln Ser995 1000 1005Ile Thr Arg Leu Leu Lys Asn Ile Thr Ala Arg Phe Val Leu Ile His1010 1015 1020SUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16WO 98/13501 PCT/US97l 16718-120-Ser Pro Asn Pro Met Leu Lys Gly Leu Phe His Asp Asp Ser Lys Glu1025 1030 1035 1040Glu Asp Glu Gly Leu Ala Ala Phe Leu Met Asp Arg His Ile Ile Val1045 1050 1055Pro Arg Ala Ala His Glu Ile Leu Asp His Ser Val Thr Gly Ala Arg1060 1065 1070Glu Ser Ile Ala Gly Met Leu Asp Thr Thr Lys Gly Leu Ile Arg Ala1075 1080 1085Ser Met Arg Lys Gly Gly Leu Thr Ser Arg Val Ile Thr Arg Leu Ser1090 1095 1100Asn Tyr Asp Tyr Glu Gln Phe Arg Ala Gly Met Val Leu Leu Thr Gly1105 1110 1115 1120Arg Lys Arg Asn Val Leu Ile Asp Lys Glu Ser Cys Ser Val Gln Leu1125 1130 1135Ala Arg Ala Leu Arg Ser His Met Trp Ala Arg Leu Ala Arg Gly Arg1140 1145 1150Pro Ile Tyr Gly Leu Glu Val Pro Asp Val Leu Glu Ser Met Arg Gly1155 1160 1165His Leu Ile Arg Arg His Glu Thr Cys Val Ile Cys Glu Cys Gly Ser1170 1175 1180val Asn Tyr Gly Trp Phe Phe Val Pro Ser Gly Cys Gln Leu Asp Asp1185 1190 1195 1200Ile Asp Lys Glu Thr Ser Ser Leu Arg Val Pro Tyr Ile Gly Ser Thr1205 1210 1215Thr Asp Glu Arg Thr Asp Met Lys Leu Ala Phe Val Arg Ala Pro Ser1220 1225 1230Arg Ser Leu Arg Ser Ala Val Arg Ile Ala Thr Val Tyr Ser Trp Ala1235 1240 1245Tyr Gly Asp Asp Asp Ser Ser Trp Asn Glu Ala Trp Leu Leu Ala Arg1250 1255 1260Gln Arg Ala Asn Val Ser Leu Glu Glu Leu Arg Val Ile Thr Pro Ile1265 1270 1275 1280Ser Thr Ser Thr Asn Leu Ala His Arg Leu Arg Asp Arg Ser Thr Gln1285 1290 1295SUBSTITUTE SHEET (RULE 26)?W0 98l1350lCA 02265554 1999-03-16121Val Lys Tyr Ser Gly Thr Ser Leu Val Arg Val Ala Arg Tyr1300 1305Ile Ser Asn Asp Asn Leu Ser Phe Val Ile1315 1320Ser Asp Lys Lys1325Thr Asn Phe Ile Tyr Gln Gln Gly Met Leu1330 1335Leu Gly Leu Gly1340Glu Thr Leu Phe Arg Leu Glu Lys Asp Thr1345 1350Gly Ser Ser Asn1355Leu His Leu His Val Glu Thr Asp Cys Cys Val Ile Pro Met1365 1370His Pro Arg Ile Pro Ser Ser Arg Lys Leu Glu Leu Arg Ala1380 1385Cys Thr Asn Pro Leu Ile Tyr Asp Asn Ala Pro Leu1395 1400Ile Asp1405Ala Thr Arg Leu Tyr Thr Gln Ser His Arg Arg His Leu Val1410 1415 1420Val Thr Trp Ser Thr Pro Gln Leu Tyr His Ile Leu Ala1425 1430 1435LysAla Leu Ser Met Ile Asp Leu Val Thr Lys Phe Glu Lys1445 1450AspAsn Glu Ile Ser Ala Leu Ile Gly Asp Asp Asp Ile Asn Ser1460 1465Thr Glu Phe Leu Leu Ile Glu Pro Arg Leu Phe Thr Ile Tyr1475 1480 1485Gln Cys Ala Ala Ile Asn Trp Ala Phe Asp Val His Tyr His1490 1495 1500Ser Gly Lys Tyr Gln Met Gly Glu Leu Leu Ser Ser Phe Leu1505 1510 1515Met Ser Lys Gly Val Phe Lys Val Leu Val Asn Ala Leu Ser1525 1530Lys Ile Tyr Lys Lys Phe Trp His Cys Gly Ile Ile Glu Pro1540 1545 1550Gly Pro1555 1560 1565Val Tyr Thr Cys Tyr Met Thr Tyr Leu Asp Leu Leu Leu AsnSUBSTITUTE SHEET (RULE 26)..........._..—.—...—............... . _. .,.,‘.....13901470PCT/US97/16718Thr Thr1310Val AspVal LeuThr Val1360Ile Asp1375Glu LeuArg AspGlu PheSer Thr1440His Met1455Phe IleLeu GlyArg ProSer Arg1520His Pro1535Ile HisSer Leu Asp Ala Gln Asn Leu His Thr Thr Val Cys Asn MetGlu Glu?CA 02265554 1999-03-16W0 98/ 13501 PCT/US97/ 16718-122-1570 1575 1580Leu Glu Glu Phe Thr Phe Leu Leu Cys Glu Ser Asp Glu Asp Val Val1585 1590 1595 1600Pro Asp Arg Phe Asp Asn Ile Gln Ala Lys His Leu Cys Val Leu Ala1605 1610 1615Asp Leu Tyr Cys Gln Pro Gly Thr Cys Pro Pro Ile Arg Gly Leu Arg1620 1625 1630Pro Val Glu Lys Cys Ala Val Leu Thr Asp His Ile Lys Ala Glu Ala1635 1640 1645Arg Leu Ser Pro Ala Gly Ser Ser Trp Asn Ile Asn Pro Ile Ile Val1650 _ 1655 1660Asp His Tyr Ser Cys Ser Leu Thr Tyr Leu Arg Arg Gly Ser Ile Lys1665 1670 1675 1680Gln Ile Arg Leu Arg Val Asp Pro Gly Phe Ile Phe Asp Ala Leu Ala1685 1690 1695Glu Val Asn Val Ser Gln Pro Lys Ile Gly Ser Asn Asn Ile Ser Asn1700 1705 1710Met Ser Ile Lys Asp Phe Arg Pro Pro His Asp Asp Val Ala Lys Leu1715 1720 1725Leu Lys Asp Ile Asn Thr Ser Lys His Asn Leu Pro Ile Ser Gly Gly1730 1735 1740Asn Leu Ala Asn Tyr Glu Ile His Ala Phe Arg Arg Ile Gly Leu Asn1745 1750 1755 1760Ser Ser Ala Cys Tyr Lys Ala Val Glu Ile Ser Thr Leu Ile Arg Arg1765 1770 1775Cys Leu Glu Pro Gly Glu Asp Gly Leu Phe Leu Gly Glu Gly Ser Gly1780 1785 1790Ser Met Leu Ile Thr Tyr Lys Glu Ile Leu Lys Leu Asn Lys Cys Phe1795 1800 1805Tyr Asn Ser Gly Val Ser Ala Asn Ser Arg Ser Gly Gln Arg Glu Leu1810 1815 1820Ala Pro Tyr Pro Ser Glu Val Gly Leu Val Glu His Arg Met Gly Val1825 1830 1835 1840Gly Asn Ile Val Lys Val Leu Phe Asn Gly Arg Pro Glu Val Thr Trp1845 1850 1855SUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/13501 PCT/U S97! 16718-123-Val Gly Ser Val Asp Cys Phe Asn Tyr Ile Val Ser Asn Ile Pro Thr1860 1865 1870Ser Ser Val Gly Phe Ile His Ser Asp Ile Glu Thr Leu Pro Asn Lys1875 1880 1885Asp Thr Ile Glu Lys Leu Glu Glu Leu Ala Ala Ile Leu Ser Met Ala1890 1895 1900Leu Leu Leu Gly Lys Ile Gly Ser Ile Leu Val Ile Lys Leu Met Pro1905 1910 1915 1920Phe Ser Gly Asp Phe Val Gln Gly Phe Ile Ser Tyr Val Gly Ser His1925 1930 1935Tyr Arg Glu Val Asn Leu Val Tyr Pro Arg Tyr Ser Asn Phe Ile Ser1940 1945 1950Th: Glu Ser Tyr Leu Val Met Thr Asp Leu Lys Ala Asn Arg Leu Mot1955 1960 1965Asn Pro Glu Lys Ile Lys Gln Gln Ile Ile Glu Ser Ser Val Arg Thr1970 1975 1980Ser Pro Gly Leu Ile Gly His Ile Leu Ser Ile Lys Gln Leu Ser Cys1985 1990 1995 2000Ile Gln Ala Ile Val Gly Asp Ala Val Ser Arg Gly Asp Ile Asn Pro2005 2010 2015Thr Leu Lys Lys Leu Thr Pro Ile Glu Gln Val Leu Ile Asn Cys Gly2020 2025 2030Leu Ala Ile Asn Gly Pro Lys Leu Cys Lys Glu Leu Ile His His Asp2035 2040 2045Val Ala Ser Gly Gln Asp Gly Leu Leu Asn Ser Ile Leu Ile Leu Tyr2050 2055 2060Arg Glu Leu Ala Arg Phe Lys Asp Asn Gln Arg Ser Gln Gln Gly Met2065 2070 2075 2080Phe His Ala Tyr Pro Val Leu Val Ser Ser Arg Gln Arg Glu Leu Ile2085 2090 2095Ser Arg Ile Thr Arg Lys Phe Trp Gly His Ile Leu Leu Tyr Ser Gly2100 2105 2110Asn Arg Lys Leu Ile Asn Lys Phe Ila Gln Asn Leu Lys Ser Gly Tyr2115 2120 2125SUBSTWUTE SHEET (RULE 26)?W0 98/13501Leu Ile Leu2130Ser Glu Lys2145Phe Lys ValTyr Ser AlaCA21502165Leu Ile Lys Asp(2) INFORMATION FOR SEQ ID NO:5:-124-02265554 1999-03-1621402155Thr Val Lys Glu Thr Lys Glu Trp Tyr Lys2170(i) SEQUENCE CHARACTERISTICS:(A)(B)(C)(D)TYPE:LENGTH: 15894 base pairsnucleic acidSTRANDEDNESS:TOPOLOGY: linearsingle(ii) MOLECULE TYPE: DNA (genomic)(xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:ACCAAACAAA GTTGGGTAAG GATAGATCAA TCAATGATCATCAAGATCCTTAAGGAGCTTGTGGAGCCATTTACCACTCGGCGGGCCCAAGTCAATTGATTCCAGAGTGAATGAGGCGGAGATGGTTCGATGATTCTGGGCAGACACGGCTAGTTGGTGAATTATCAGGGAGCATTGTTCCAGAGGAATCATCCAGACTAACTAACAGGGTCAGAGGATCCCAGTCACAACCAATACTTTGAACAAGGAATACCATTCTAAGCTGATTCGATTCAGATTGACAAGAGCAGAAAAGAAACAAAACACATTACTGGACCGGTGCACTAATAGACCGATGACCTCTGGCCTTATCACATGATGATCTCAGATAGCCCAAATTTGAGCTAAGAAGAGAGAAAATGATTAGGGATAGGACAAACCTTATAGTACCTGGTCAGGTTGTATATTATCCTGACGTTAGCCTTCGCATCATCCAAGTAGTTGAAGTGCAGGGTCTTGCTGGTGGATAAAGGTTGGATGTTATTCTAGTAATCCGAGATGACCCATTACAAATCCCTGGAAATTGGAAACCTTGTTTGTGCATCAGGCTGAAGAGGTACCTAGTGATCAAAGACCCTGAGCGCGAAGGCGGTACACCCAAGGTGAGGAACSUBSTITUTE SHEET (RULE 26)PCT/US97/ 167182175CACTTAGGATGCCACACTTTTCAGGATCCGGATTCCTCAACCGGATGTGAGAGTCTCCAGTTAGAGGTTGAACATGGAGGTCCAGGTCCGGGATTCAACAGTTACGGCCCCAAAGAAGGGAGGATTGCCGAsp Leu His Gln Asn Ile Phe Val Lys Asn Leu Ser Lys2135Gln Ile Ile Met Thr Gly Gly Leu Lys Arg Glu Trp Val2160Leu Val Gly60120180240300360420480540600660720780?W0 98/13501AGGACCTCTCGGAACAAACCGATTAGCCAGGACTGCATGAAAATGGGAGAGTGCAGGATCACTCCATGGGGGCAAGAGATGTATCACTGCGGACCAGTAGGTGAGAATGAGAGAAGCCAG_cccarcrrccCGCAGGACAGTGGAAGAACAACTAGGTGCGAAAACTTAGGGGGCCGATGGCTCAAGGCCGATATCAGACAGGTCTCAGCATGCGGTCAGGAATCTCCAGGGCGGTTAAGGAGCACCCTCTGATACCGAGGCTTACGCCGAAAGGATTGCTTTTTATCCTAATTTGCTGGTAACTGCACCCATACCCCCTGAGGTTTGAACGGTGAGGAGGTGAGGATGCAAGCGGTTGGAGCTACCAGGAGGAGAGCTACAACCAGCGCATCGACGGTCAAGGCTCAGACAGAGGCCGAGAACCAGGTCCCAGAAGAGCAAGCCCATCGGACCCAGGACAAACCATGCCTGATCTGGAGACATCAAGCACGAATCCAAGACAGGAGGAGAGATATGCTATCATTCATGGTCGGAAATGATATACTATTAAGTGAGTTATCCATACATGGTAACTCTGGAGCTTTTGGTCGATTCAGCTGGGAAGGCTTGTTTCCCAGACAAGTTGGGGGGCAAGAGAAACCGCCCCTAGACAGCTGACGCCCACGGACACCCGACCAGAACAACACAGCCGCGGCACGCCATCTCACTGGCCGGACCGAACCCTCAGCAATTGAGCGATGACTGGGTTACAGTGCTGACTCTCGATGAATCTCACTGACCGG- 125 —CTCTAATCCTGTGACATTGATTGGGATAGACACTTGAGTCTCCTGGAGAAATGCCATGGGCTTACTTTGAAAGTCAGTTCCAGAGATTGCCCCAAGTGTCAGGAAGATAGGGTCTAGCAGTTGACACTGCTGCTCAGGCTCTAGGGTGTAACATCCGCCTCAGCCAACCAGTCAAAAACGGTCGAGGAAGACCCGCAAGGGGATCAACTGAACGCTGEAATGTTATCATGATCATGGTTCGAAAACAGCGGGATCTGCTC02265554 1999-03-16GGATATCAAGTACATATATCAACTATGTATCTTGATGAATCTCAATTCAGAGTAGGGGTGTCCAGCATATCACATTAGCAAATGCACACTATTTCTACACGAGGGTCAAAAGCAAGCGATATCGGAGTCAGCAAGCCATGCAATGACAGAACCCTCCATCACCATCCACTGACTGGAATGCCATGGCAGCAAGAGGAGGCAAGGCAGTGCCTTTGGGAATTTTATGATCAAATCAGGCCTATGTGGATATCCATCTCTATSUBSTITUTE SHEET (RULE 26)PCT/US97/16718AGGACACCCG 840GTAGAGGCAG 900ccrccrcrrs 960CTTTACCAGC 1020AACAAGTTCA 1080GAACTTGAAA 1140TTTAGATTAG 1200TCTGAACTCG 1260ACTGAGGACA 1320GGTGATCAAA 1380CAGAGTCGGG 1440GCGAGAGCTG 1500GGCCAAGATC 1560GCAGGAATCT 1620GATCTTCTAG 1680ATTGTTATAA 1740CCTACGACTG 1800CATCCGGGCT 1860ATGGTCACAA 1920AGGCAGTTCG 1980ACCTCGCATC 2040CCCCTCAAGA 2100CAGCGGTGAA 2160TGATGGTGAT 2220TGGCGAACCT 2280GGGGTTCAGG 2340?W0 98/ 13501GCTTCTGATGAGAGGCAACAGGTAGGGCCATTTGGAGCGGCCCTCGGAACGCCGCACTGAAATAATGAAGAAAACAGCCTCTGCTGTTATAGCATATCCAAAGGATCCCAGGCAGAGATTCTCCAAGGAACTAAAGCCGAGCATCACGCACGTTACCTGACAGATGCTGACCAATCGACCGCCTCCCAAGAAGGGTCGATTCAGAGTCATTGCTGGGGGTCCCTGCCCTTCTGAGCTTGAACAACACCCCTCAACGCAAATTGAAACTGCACTTCCCGAAGCACTTCCGAAGATCGCGTCCATCAGGGCCTACAGGAGTGAAGGGGGAGATGGCCAAAATTGAAGGGAGACCCTGGAAGGACGACCCCACCAGGCCGAGCTGACAAATGGTCGGGAAAAAGTGTAATCCGTGACTCTCCTTGAAGATAATTAATTAGTACTTCCACAATGCGCTCCGATAAGATCCTGGTTGTTGAGGACAGGTGTTGGTCATAGTCGTTACTAACTCTCCCAAGTGTGCCAAGAAGGAGGGGCTTGGGAAAGACACCCATTTTTATTGACAAGGTGCACCTGACACCCGAATTATTATGATACACGAGGATAGTTGAGTCAACACCTCTCATGCAGATGTCACTGGCCGAAACGGACCAGTGATGAGCTCACTCCATTATATGATGATATCAATGAAGTAGAGCCTAAATCACAGAGATCTCAACCTACCACTAGGCGACAAGCGATCCCCAGATCCACAGAGACGTACAGCTCACACCTTAATGCGGTTA-125-GAGATCCACGACTCTCAATGAAAAAGGGGAGGTGGTGCAAGTGGGGAATGTCTGGTACCAGATGAGCTGTAATCAGAAGAATTAAAAAGCAGCATCATGAGAACTCAATCGTTCTCAAGATCCAGAGGACGCCGTCGGGTAAATCCAGCCAAAGGAGCCACTACAGCTCACATTATAAAAACGACTTCGACCTACAGTGAGGAAGGATGATAGGGCCTCCCAAAACCCGACAGGGCTCAAGGAGAAAGGTATCTGATACC02265554 1999-03-16AGCTCCTGAGTTCCTCCGCCCAGACGCGAGCCCAATGTGCTCCCCGAGTGCAATCTCCCCTCTCCGATGTTAATCTCCAAAGATCAACAGTCGCCATTCCCCGACCTGAAAACCCGTTGCAGCTGCTGAATTGTTCCTGAGGCTAGAGGAACGATCTTGCACTTACCTGCAACTTAGGAGCAAGTCGGCATGGCAGGCTGATGCTTTACGAATCGGGCGAAGAACTCCTCTGAAAAACTGCCTAACAACAGCTGGATACCSUBSTITUTE SHEET (RULE 26)PCT/US97/ 16718ACTCCAATCT 2400CCCGAACCCC 2460ATTAGCCTCA 2520TCGAAAGTCA 2580TGTGAGCAAT 2640GAGATCCCAG 2700CCAAGACATC 2760GCTAGAATCA 2820GCAAAATATC 2880TGGACTTGGG 2940ACCCATCATA 3000CAGCCGACAA 3060GGAATTTCAA 3120CACCGGCCCC 3180GGATCGGAAG 3240CAAGTTCCAC 3300CAACCTCATG 3360CAAAGTGATT 3420TGGGACATCA 3480GTGCCCCAGG 3540TACATGTTTC 3600GCATTTGGGT 3660AAAGAGGCCA 3720GTGTTCTACA 3780GGGAGTGTCT 3840CCGCAGAGGT 3900?WO 98/13501TCCGTGTTGTGAAGAATGCTGGATTGACAACAACATTTATATTATTGCAAGCACCAGTCTGGTTCAAAAATCTGGAGGAGAAGAATTCCGTGTAGACCGTGCCCGGACAAGCTGACGGCACCACCAGCCACGCCCCCGACACTGGAAGGCGATCGAGGTGACTAAACAAACGGCGCCGCGCCCCGGTGCCAATTCAAGACGAGGAAGCCCGGGCCACCAGACCCCTGCCCCGAAGGGCCCCTCCCCCTCTCCCACCCCTATTATATGAGCAGAATTCAGAAGCGATTGGCGGTCCACATCAATGAAAATCTCACATTAGAGACCTTATGTCAGATGCAAGCATTTACGACAGTGCCCAGCAAAAGCCCCCAGCGTGAACATCCCAATCTGCCAGACCACCCCCTCCCCCTACCCAACCGCACTTAGGGCCCCCCCACCTCCACAGGCAGGGGGGGGCCCCACCCACCCCATTCCCAGACTTGATCCGGTGCCGAACCGCATCTCGAAGGGAAGGAGACACCAATCACCCGTCTCGGTCAATGCCTGGGAAGAGGGAACTTCAGAAAAGATGGAGCACAGGCATACCCACTGAATAGTAAGAAGACGTGATCAAATACCCGAATCCAAAAGACCCAGGCGGCCCGTCCTCCTCAACCGCATCCTTCCCTCAACAGGCATCCGAAAGGAACATACCGACAACCACACACCAACCCCCCAAAAAACACACGACCACGGCCATCACGGCGGCCACCAAAGACATCAACCAAAAGATCGGGAATCCC-127-TTTCGGATAACAGTGGCTTTTCATCGATAAGGAGAAAGAAGCCTGGTTTTAAATGAGCAATGGATATCAATCCAGGCAGTTAAATGATGAAACGACCCCCTCCACGGQCCTGGGCAC%GAGTGGGACCCCCCACAGCCCCGCAAGAACTCCTCCCTAGACCACACCCGACGAGGGAGCCCCTCGAACAGAAGGCCCCCAGCAGGAACCGACCCGCAGAAACAACCCGAACGTATCCCACACAATCCACCAAGAATCAAGA02265554 1999-03-16CGGGTATTACCAACCTGCTGTGCAGAGCAAGAGTGAAGTCTGCACTTGGTGACTCTCCATTGAAGACCTTTTTGCAGCCACCAAGGACTACTCATAATGAAAGTGAGAGGACAGCCCCGACGAGGACCAACGGGAAAGAGCACAACCGAAAGATCCTCTCAGAACCCAGACCAACCAATCCCCAGCACCCGGGCCGACAGACCAGAATCCGGAAAGGCCACAGCACCCAAGCCTCTCCAACACCCGACGACTCATCCAATSUBSTITUTE SHEET (RULE 26) PCT/U S97/ 16718ACCGTTCCTA 3960GTGACCCTTA 4020CTTCCTGAGG 4080TACTCTGCTG 4140GGGATAGGGG 4200GCACAACTCG 4260AATCGATTAC 4320TCAGTTCCCC 4380TTCAAAGTTC 4440CAGCCAGAAG 4500CCAGCCAGCA 4560CACAAGGCAA 4620CCCCCAAGGT 4680ACCCCCAGCA 4740CCGCACAAGC 4800CCCCCGGCAA 4860CCCCGGCCCA 4920CCGCCGGCTC 4980AGCCATCGAC 5040CCAGCACCGC 5100AGACCACCCT 5160CAACCCGCGC 5220GAGCGATCCC 5280GTCCCCCGGT 5340CACTCAATTC 5400GTCCATCATG 5460?W0 98/ 13501GGTCTCAAGGACCGGTCAAAAGCTACAAAGATAACTCTCCACAGTTTTGGCAGAGTGTAGGCCCTAGGCGwrcnncwcwcGAGGCAATCAATCAATAATGCTAGGGCTCACGGGACCCCA_ATCAATAAGGAGCAGAGGAAAGTATAGCCTGTCTCGTACAcnacccraccGTGTGCAGCCTCCACCAAGTTCACAAGGGAACGATCATTAGTGGTCGAGGTACTTGCACAAATCTGGGGACAGATATTGAGTGTGTCTTGTGAACGTCTCTCCATTGGGGTTATGACTCGTCAATAACTGAACCAATTAGCTTCAAGTAGTTGCCACAGCAAGCCATCGAGACAAGCAGGAGCTGATACCAATTGCTCAGTATCTGCGGATGTTAGAAAATAAAGGCCCGATCCGACGCTACATAGGCTCTTATCTCGAAAAAATGCCTTCCTGTGCTCGATCTAATAGCATCAGGACCCTGAACGGCGTGAATTGACCTATGCAATTGCGGAGTATGAAGAGGGTTGATCATGCCATATTCCAATCTCTCTTTCCAGCCATCACGAGGGTAAGATGCACTTGAGACACAAGTGCTCAGATACAATCTGAGAGCAGGAGATGGTCTATGAACATACTATACAGATATCTATCGCTCGGATACGATAACTCACGTCCGAGATTTCAAGAGTGGTTTTGATGAGGTACCCGATGTACACTTGTACAATTGTGCATGACAAGATCGACCATCCAACGGTCCTCCCTAAGTTGGAGAGGTTTATCGAGGGATCCCC-128-ATGGCAGTACAAGATAGGGGCAATCATTAGGAGATTGCAGAATGCAATGAAGATTTGCTGACAGCCGGCAGCGAGCCTGGATATTGGCTGCAACTATCTTGAAATCCTGTCAGGCTTTGAAGTGGAGGTGGTCGACACAGAAGGGGGTGATATACCACTGTCATCGTGTAAGTCCTCTGCTCCGGGTCTTTCAATCCTTTCTAACATACAGTCGGGAGCAATATCATTGGGATGCCAAGGAGCACTAGCAGCTTTAATAT02265554 1999-03-16TGTTAACTCTTGGTAGGGATTCATAAAATTAATACAGGAGCCCAGAATATGAGTTGTCCTTTGCACTTCAAAACTACTAATTCAGGGTGTGTGATTTAATCACTATTTGGGCTATGCGCTATTTACTGGGAGTCCTACTTTTGTCCACCGTGCCCAAGTACTTTCATGCCTCCAAGAATGTTGGGAACCGGCAAGTGTTATTGCTGCCGAGGCGGTATCCAGAGGTTGGAAATTGTTGGATAGTTTACATGTTGCTGCAGSUBSHTUTESHEET(RULE25)PCT/US97/16718CCAAACACCC 5520AGGAAGTGCA 5580AATGCCCAAT 5640ACTACTGAGA 5700AAGACCGGTT 5760GGCGGGTGCG S820CCAGTCCATG 5880TCAGGCAATT 5940CCAAGACTAC 6000CGGCCAGAAG 6060CCCCAGCTTA 6120TGGAGGAGAT 6180CATCTTAGAG 6240CATTGTACTC 6300GCTAGAAGGG 6360TGTTGCAACC 6420AGAGGGGACT 6480ccrccscsss 6540GTTCATTTTA 6600CACAACAGGA 6660TCACTGCCCG 6720GGACGCTGTG 6780CGTAGGGACA 6840GTCATCGGAC 6900CCTGATTGCA 6960sccsccrrsr 7020?WO 98/13501AACAAAAAGGACATCAAAATCACAAGTCTCTGTCTCCGGAATCATCCACATCCTAGGGGATTTGCTGGCTCATAAGACTTTCTAGATGTAGATCATCGGTCATCTCTGACTTGGTGTATCGGCTGCTGAACAATCAGTTCATTCTCAAACATCTATAGTCTAATCTGAGCAGGTGTTATCGCAACCAGTCAGCCCTTTGTCAGCTTCCAGCCCCCTATCATATCGCTGACAATGGAGACACGAGTGGGCAGAGTCTGACAGAGAACAAGTCCTATGTAAGCTCTTCGTCATTCCCTCTGGATGTCACCACAGTAGGATAGGTTCTATTCGCATCGGGCAGACTAACTCAAGATGAAGTGGAAGATTAAATAACCCGCCAGGAACTCATGACTAGCTGTCTATGTCGCTGTACTATGACATAGTAAAGGGTAGAAATCCGGAGTAATGATTCACAGGGAAGCTCGTCAAGCACGGATGATCAATCAAGCAATGCTTCCAGCCCATTGAAGGGTTGAGCTTACATGGTATGTCAGTCGCTCTGATCAAGCAACCCCGAACAATAAACGAGACCGTTATTAACAGTCATGTTTCTCCATCTACACTCGAGCATCAGCCTGAGGACTCCTTAATCCAGAGAATCAAATGCATTGGTCAAAGGGAAACCCTGTTGGACCCAGGGAATCAGAGTTGTCGTTTGGGGGCTCAGCAACTGATTCTATCACTAGGTGTCTGCAGTGATAGAAATGGGCTGTAGGCGTGTAAATAACAGGATAAATCAAAAT-129-AGACCAGGCCTCCTCTACAAACCGCATCCATCGGTAGTTAGATAAATGCCAGAACATCTTGAGCTTGATCCGCAGAGATCGGTCAAGGACACCTCAGAGAGGATAGGGAGATTGGATTATGAACTCAACTCTGCTCAGGGCTTGTATTTAGTACGGGGGAACAACTGAGCTCCGGTGTTCCATGGTGGCTAATTCCCTATGAAATCCCCACAGGCTCTACCCCGACAACAGGGTAAAATCTCCTTCATACTGCTTCAGGA02265554 1999-03-16TAAAGCCTGACTCTTGAAACGCATCGAGCCATTAAAACTTTTCTACAAAGATGATTGATAGGGTTGCTAGCATAAAAGCCGTGCTGACACTTCACCGACCTACGACTTCAGATCAATACTCTACTGGAGGCCCACTACAAAATCGAGGTTACTTACCTAGATGCACCGAGCATATGACAATTGGGGGAGCCAGGGATCAGACCGACATGCCTCTCATCTCCGGACAGATGCAAGCACTCTGGGGTCTTGTTTCGGGCCATSUBSTITUTE SHEET (RULE 26)PCTIU S97/ 16718TCTTACAGGA 7080ACAAATGTCC 7140CACCTGAAAT 7200AGGGTGCAAG 7260ACAACCCCCA 7320GACCTTATGT 7380CCATTGCAGG 7440TCAGCACCAA 7500CACTCTTCAA 7560TAGTGAAATT 7620GAGATCTCAC 7680GTGCAGATGT 7740CCAGGGTAAC 7800TCAGAGGTCA 7860ACAATGTGTC 7920TGGAAAAGCC 7980TGTTTGAAGT 8040ACTATTTTGA 8100TCAAATTCGC 8160GGAAAGGTGT 8220AATCCTGGGT 8280ACAGAGGCGT 8340ACAAGTTGCG 8400GCGAGAATCC 8460CTGTTAATCT 8520TGATCACACA 8580?W0 98/13501CGGTTCAGGGGCCAATGAAGGGTTAGTCCCAACATACCTAACCTGGTCAATGTGGTTTATGCCTATAAGGCTGGTGCCGTTGGGATGGTGATAGGGCTGCGTGAAATAGACGCTATCTGTATAAGATAGTCTACACTGTGTAAACAATGTCTCATATTCCCAAGGAAGATAGGTTTTCCAAGGACATCAAAGCCCTTTCTCCCATACTTGTGCTAATCTCTGACGTTTGACCGCTATGACAACTGATAGATGGAGCCTCTATGGACCTATAACCTAGCCTTACCTCTTCACCTGCGGAGGGATCTCCAATTACGTTTACAGGGGTCCCCACACTTCTGTGGGCATGGGAGCAGTGAACCACATCAGAATTCAACCAGATCAGCTATCCTGTCAGAACATCGGAAGTTGGGATATCCAAATCCGTGAGCTCATGCCTGAGGGGAGAAAATTGTTTTGGTTTCCATAGGAGGTCGTGACCTTACTGGTCTTGCATTGATGCTTGGTTTCTTCTTCACTTGCTCAACAAATCCAATAGGTGTAATCTGTTCCAATTGGATGGTGAATGTTTTGGCGCCCAAGCCGTCGAATTACATGCTTGCGGATCAGCTGCACATCACATGATAAGAAAAACGTTATACCCTGGAGTATGCTCAAGCACCGCCAATGTCATCATGTAATCAGGCTCAAAAAGGGACACTAACTATTAACTTGGACAGTCAAGAAGACACACACGTTGCTATAAATGTATTGTGAGGTATACAGCCTGCACTCGTACCTGCAGC02265554 1999-03-16PCT/US97/16718- 130 -CCACAACAAT GTGTATTGGC TGACTATCCC 8640CAACACATTG GAGTGGATAC CGAGATTCAA 8700TAAGGAAGCA GGCGAGGACT GCCATGCCCC 8760TGTCAAACTC AGTTCCAATC TGGTGATTCT 8820AACCTATGAT ACTTCCAGAG TTGAACATGC 8880CTCALLLLLI TALLLLLATC LLLLLAGGTT 8940AGTGGAATGC TTCACATGGG ACCAAAAACT 9000CTCAGAATCT GGTGGATATA TCACTCACTC 9060AGTCACTCGG GAAGATGGAA CCAACCGCAG 9120GTCACCCAGA CATCAGGCAT ACCCACTAGT 9180TAGGGTCCAA GTGGTTCCCC GTTATGGACT 9240AAGTTCACCT AGATAGCCCG ATAGTTACCA 9300GAGTCCCTCA CGCATACAGC CTGGAGGACC 9360TAAAAAACGG ATTTTCCAAC CAAATGATTA 9420AGTCCAAGCT TAGGAGTTAT CCGACCCACT 9480ATTTATTTAA CATAGAAGAC AAAGAGTCAA 9540GAAATTCGCT GTACTCCAAA GTCAGTGATA 9600CACGGCTTGG CCTAGGCTCC GAATTGAGGG 9660GAGTTTACAT GCACAGCTCC CAATGGTTTG 9720CTGAGATGAG GTCAGTGATT AAATCACAAA 9780CAGTATTCTT CACTGGTAGT TCAGTTGAGT 9840TCAGTAAAGA GTCTCAACAT GTATATTACC 9900ATGTCATAGA GGGGAGGTTA ATGACAGAGA 9960AGCTTCTAGG AAGAGTCAGA TACATGTGGA 10020GGAATCCAAC TTACCAAATT GTAGCCATGC 10080TGAGGGATAT AACAGTAGAA CTCAGAGGTG 10140SUBSTITUTE SHEET (RULE 26)?WO 98/13501CTTTCCTTAAATGAAGGTACTACATCTGACCAGTAACGGCAGACTCTGATGGCACGGAGGATGCTCAAGCTTGCTGGAGTACCTAAAGGAAGTTCCTGCGTTAATGATTCTCCATGACCCGTAGACTTTTTAATCTCAAAATTTGACTAAGTCACAGGGGAGAACGTGAGACACTGATCAATCTCAAGAATAAATGAGATCTGTCCTCTAGCAAAGTCCCGTCAGAAGCTGAGTAAGGATTACCCAGCACACTTTGTAATCCACTGCTTTTTATCATGAGAGGGGAGATTTGCTGAAAATGAAAGGTCATCAGTTGGCCATTCAGGTGAAGAAATTTGGCCAAGGCACTTTTACGACCCTGAGCTTTGACTGAGTTCAACTGCTAAAATGCGGGATTGGCGGCACTCCACGGGGCCAGTCAGCAGCAAAATCCGGAGAATGTACTGCCTTTTACGGATTATGTAAGTGACCAATGACCAAGTGGACCATCTGCTTCGTTAATGGCCTTACTCTTAGGCAACAACTGAAATACTTAATTGAAGTTCTCATTTTGTTAGGAAATGCCATATTCTCCCCTGACCCGGGTTAACACTGCTTTATGCGCTGCTCTCCCCCAAAGGAACCATATGACACTGTCTTACAACTTACAAAAAATTATTTTAACTCTAGCTGTTAAAAACCCGGGTTTATAGATGGAGGCTTAATTGGAGATCCCTCATTTTCCTCATTGCCATCTTCATTAAGCACCATTCGTGCAAGGGGAACCTTAAGAAGGCTACATG-131-ATGATGTTCTCCCTAGATTATCAGAAGTTTACATGAATCAGTGGAATCATTCCCCCTGCAATGAGCAGTGCTCTTAGCCTAAAGGGAATGCTGGGTCACGTGATAATGTAGCCTGAAAGATGAGGGCATGAGGACAATGGTCTCAGGAGTACTCCCGAAGGATTCCCTCAACGAGACRGTATGAGACCATTCCAGTGGCTCCCCTGACCTAGTACCCTATCCTATTTATAACAATCAGACAACGGGAAGCACATAGGCCA02265554 1999-03-16TGACCAAAACCATTTTCATACGGCCACCCCGCCTAAAGTCAATCAACGGCTGCTGCAGACCGTTGATAACGGATAGTGATGGATTCAGTTGAGGCTTGTATGTTGTAAGTAAAGGAGATCCCAAGTGATTGATGGCCAAGCCCCAAAGATCCCAGTCCACTGTAATTCGGCAGTGCATTTCAGCTTATTTGCATAAGAGGTGACGCCCATGGGAGGTATACCTGGCTGCTCATAGCCGTATGCTAGAGTATCACCTCAAGSUBSTITUTE SHEET (IRULE 25)PCT /US97/16718GGGTTTTCTG 10200ACTGATGACA 10260AGACTTGAAG 10320ATTGTGTATG 10380TATCGTGACA 10440ACAATCCGGA 10500TGGAAATCTT 10560CTGACAATGT 10620TACCCGAAAG 10680AATGTTTTCC 10740GGAGCTTACC 10800AAGGAAACAG 10860GCTGAAAATC 10920GACGAGCACG 10980CTCAAAGAAA 11040ACAAGTACCA 11100CAGGACCAAG 11160ATCACGACTG 11220GCACAAAGGC 11280CTTGAAACCT 11340GTCCCGTTAT 11400GAAGGGTATT 11460TATGAGAGCG 11520ACAAAAAGGG 11580ACTAGAGATT 11640GCAAATGAGA 11700?W0 98/ 13501CAATTGTCTCTGTCCCAATCAAACAAGGGCATGACCGTTACTCTTGGCTTACAACGATCTTGAATATGAGATCTCAAGAGCACAACAACCTTGTATGCGTTCCATAGTCCAGGGACTGGCTCCTGGATCAAAGGCCTGATTGTCCAATTAGAAATGTCCTATATGTGGGCTAGAATCTATGATCAGTCAAAGGAAACATCTGAAGCTTGCCAGTGTACTCCAAGGCAAAGCGACTAATTTCCCTTGTCCGCAGATAAGAAATCACATTTTACTCAAGAGCAGCATGCAGTCCTTGCATATCACAATCAATCTTAATAAGGCAGGCTGTTTAATGATTCTCGGGGGACTCTCCAGAGCATCAAACCCAATGGGCATTCCTCTAGTGTCACATCGAGCCAGCTGACTATGAACATTGACAAAAAGGCTAGCTGCGAGGCCACCTACGGATGGATCCTTGAGACTTCGTAAGAATGGGCTTATGGCCAATGTGAGCGCATAGGAGTGGCAAGGGGTTGATACTCATTTGTCTATTATCGCAAGATAATATTGCTATCCCTGAACGTCAACCATGAATGGCACTGTGTCAGAAACATCATCACTAATCATTCCTAGACTAGACTCCTTAAAAGGGTATGGACAGGCGGGGCAAGAGATGAGGAAGGCAATTTAGAGGAGTCATGTTCGAGGACGGCCTTATTCGGCTTTTTTGTCCGTCCCATATAGCCCCAAGTCGGTGATGATGAGCCTGGAGGTTGAGGGATCTATACCACAAAACTTTATAT-132-CAAAAGGAATGTGTATTCTGCAACAATGGCTCCTAAAAGTCCCGGGATGTTGCCCGCTCCTCGGTGATCCTGCCTGAAGAACTGGGCTAGTCAAGAACATTATTCCATGAATATTATAGTAGTCTATTGCGGGGGTTAACCAGGGATGGTCAGTGCAGCTCTATTTACGGGCCATGAGACCCTCGGGTTGTTGGTTCTACGATCCTTGCGATAGCTCTTGAGCTAAGGGTGTACCACTCATCTCCAACGAACCAACAGGG02265554 1999-03-16ATATTATGATGTCAGAGACTTAAAAGCATCGATACAGCAAAGTCATACCCTATCGGGGGGAGTAACATCAGACCCTTCATCGACCCTTACAACTGCAAGGTGACAGTAAAACCTAGGGCAAGGCATGCTACTCTCGAGTGGCTATTGACAGGCTAGAGCCCCTTGAGGTCATGTGTCATCCCAACTGGATCACTGATGAGATCTGCTGTTGAACGAAGCCGATCACTCCCAGTGAAATACCAATCTCTCAAATGCTTCTASUBSTITUTE SHEET (RULE 26)PCT/US97/16718GGGCTACTTG 11760ATAGTTGATG 11820GAGAGAGGTT 11880ATCCTGATCT 11940CTCCTCACAA 12000ATGAATTATC 12060TCAATTGCTG 12120CAAGTAATGA 12180TCAGCAAATC 12240TTTGTCCTGA 12300GAAGAGGACG 12360GCTCATGAAA 12420GATACCACAA 12480ATAACCAGAT 12540GGAAGAAAGA 12600CTAAGAAGCC 12660CCTGATGTAC 12720TGCGAGTGTG 12780GATATTGACA 12840AGAACAGACA 12900AGAATAGCAA 12960TGGTTGTTGG 13020ATCTCAACTT 13080TCAGGTACAT 13140TTTGTCATAT 13200GGGTTGGGTG 13260?WO 98/13501TTTTAGAAACTTCACGTCGACCCGCAAGCTCTTTAATTGAAATTTGTTACCTATGATTGATAGGGGATGATCACTATCTAGACCATCAGGAAGGAGTGTTGGCATTGTGGCAACTGTGTGAAGAGTTAGAGATTCGACAAGGACCTGCCCATATCAAGGCTTGTAGACCAGATTGAGAGTCAAAGATCGGATGATGTTGCGGGGTAATCTCTTGCTACAAACGGCTTGTTAACTAAACAAAATTAGCACCTTGTCAAAGTATTGTTTCGAAACAGATTGTAGAGCTTAGGCAGAGATGCAATGGTCCACACCTGGTAACACGATATCAATCTTGGGCCAGGAAATATCAGTAAGGTGCTTTATTATAGAGCAACATGATTAGAGTTCACATATCCAGGCAACCAATTCGAAGAGGCTAGGTTACTCATGCTGATCCAGGACAGCAACAACAAAATTGCTCCGCCAATTATAGCTGTTGAGCTTGGGTGAGGTGCTTCTATCTATCCCTCCGCTLTTTAACCACTCGAGAAAGTGCGTGATCCGCAGAGCTATACAAGGCTATCCCCAACTATAAATTTGAGAAGTTTCATAATGTGCAGCCAATGGGTGAGCGTCAATGCTCCCTATCCATGTACACATGCTTTTCTTCTGTAAACACTTGTGGTCTACGACTTATCTCCAGTCTCTGACTTTTCATTTTTGATCTCAAATAAAAGATATCAGAAATCCACGATATCAACATGGGTCGGGTTAATAGTGGGGGAAGTTGGTCGGGAGGCCCG-133-ATACCGGATCCGATGATAGAGTACCAACCCACACCCAGAGATCACATTTTAGGACCATATCTGAGTTTCTTCAATTGGGCTGTTGTCTTCTAAGCCACCCGTCCTTCACTATATGACCTAGTGAAAGCGAGTGTTCTAGCCTGTAGAGAACAGGGTCTTCATCTCCGGCGACGCCCTCGCTGAGCATCAAACACAAGCAACTTTCCGCAGTAATTAGGAGCTATGTTGATTTTCCGCC$ATTGTCGAACAAAGTCACGTG02265554 1999-03-16ATCTAACACGTCATCCCAGGATTGATATATCCATAGGAGGAGCTAAGTCCGAATGAAATTGCTTATAGAGATTTGATGTAGTTCCTTTCTAAAGATCTACTGATGCTCAACCTCGACCTGCGAGGATGTAAGATTTGTACATGTGCAGTTGTGGAACATAAGGATCGATCTGAGGTAAATGGATTTCAGAGCACAATCTTAATCGGGTTAATGCCTTGAGCACTTATAAGTTCTAGATCTCAGAATGGGAGGTAGGCAGTSUBSTITUTE SHEET (RULE 26)PCT/US97/16718GTATTACATC 13320ATACCCAGCT 13380GATAATGCAC 13440CACCTTGTGG 13500ACAGCACTAT 13560TCAGCTCTCA 13620CCAAGATTAT 13680CATTATCATA 13740AGAATGAGCA 13800AAGAAATTCT 13860AACTTACACA 13920TTGTTGAATG 13980GTACCGGACA 14040TGTCAACCAG 14100CTAACCGATC 14160AATCCAATTA 14220AAACAGATAA 14280GTCAGTCAGC 14340CCTCCACACG 14400CCCATTTCAG 14460AACTCATCCG 14520CCAGGGGAAG 14580GAGATACTAA 14640GGTCAAAGGG 14700GTAGGTAATA 14760GTAGATTGCT 14820?W0 98/13501TCAATTTCATAGACCTTACCTGGCTCTGCTGGGATTTTGTTCTACCCTAGAAGCTAACCGGGACTTCACCCAATTGTGGGCTATAGAGCAAATTGATCCATCTACAGGGACTTACCCCGTTTTGGGGGCAATCTCAAGTCCTAAGTCAGATAACAATCAAATTAATTGGTTATATTAAAGAGTCAGTAATTAACAAAGATCCTTGGCAAATCAGGGATTTATACAGCAACGCTAATGAATTGGACTTATAAGACGCAGTTGGTGCTGATCCCATGATGTTGTTGGCAAGAATTGGTAAGTTATTCTTCTTCGGTTACCTGGAAACAGATTGGAGACCAAATGGACTCCGGAAAACTTTGACAATCCCTACCTACTATAGAGAATAGGATCAAATAAGTTATGTTCATATCTACCTGAAAAGAGGTCACATCCAGTAGAGGTGAATTGCGGGTGCCTCAGGGCTTCAAAGACAAGCAGGCAACTACTCCGGGAATACTAGACTATTATGACGGGAATGGTATAGACCCTAATCAAATACGAAG(2) INFORMATION FOR SEQ ID NO:6:-134-CTAGTGTGGGAGCTAGAGGATACTGGTGATTAGGGTCTTACTGAATCTTATTAAGCAGCATATCCATTAAGTATCAACCCTGGCAATTAAAAGATGGATTACCAAAGAAGGAGAACTTATACAGAAAGTTTACACCAGAAGGGGTTTAAAAGTTAGTCGGCTGCCCTAGGTTTCTATTCC(i) SEQUENCE CHARACTERISTICS:02265554 1999-03-16GTTTATCCATATTAGCAGCCTAAGCTTATGTTATAGAGAATTTAGTCATGGATAATTGAAGCAACTAAGCTATTCTGAAGCGGACCTAAAGCTTAACTCTTCAACAAGGGATCTAGGATCGATAAATCGGTATCTTCGTTACGTGAGTGGATACAGTGCCTAGTTAGGCACAGCTTTGTC(A) LENGTH: 2183 amino acids(8) TYPE: amino acid(C) STRANDEDNESS:(D) TOPOLOGY: linear(ii) MOLECULE TYPE: protein(xi) SEQUENCE DESCRIPTION:SEQ ID NO:6:SUBSTHUTESHEET(RULE25)PCT/U S97/ 16718TCAGATATAG 14880ATCTTATCGA 14940CCTTTCAGCG 15000GTGAACCTTG 15060ACAGATCTCA 15120TCATCTGTGC 15180TGCATACAAG 15240AAACTTACAC 15300CTGTGCAAAG 15360ATACTCATCC 15420ATGTTCCATG 15480ACCCGCAAAT 15540TTTATCCAGA 15600AAGAATCTAT 15660GTTTTTAAGG 15720CTGATTAAGG 15780TTATTTGCAA 15840TGGT 15894?CA 02265554 1999-03-16W0 98/13501 PCT/US97/ 16718-135-Met Asp Ser Leu Ser Val Asn Gln Ile Leu Tyr Pro Glu Val His LeuAsp Ser Pro Ila Val Thr Asn Lys Ile Val Ala Ile Leu Glu Tyr Ala20 25 30Arg Val Pro His Ala Tyr Ser Leu Glu Asp Pro Thr Leu Cys Gln Asn35 40 45Ile Lys His Arg Leu Lys Asn Gly Phe Ser Asn Gln Met Ile Ile Asn50 55 60Asn Val Glu Val Gly Asn Val Ile Lys Ser Lys Leu Arg Ser Tyr Pro65 70 75 80Thr His Ser His Ile Pro Tyr Pro Asn Cys Asn Gln Asp Leu Phe Ann85 90 95Ile Glu Asp Lys Glu Ser Thr Arg Lys Ile Arg Glu Leu Leu Lys Lys100 105 110Gly Asn Ser Leu Tyr Ser Lys Val Ser Asp Lys Val Phe Gln Cys Leu115 120 125Arg Asp Thr Asn Ser Arg Leu Gly Leu Gly Ser Glu Leu Arg Glu Asp130 135 140Ile Lys Glu Lys Ile Ile Asn Leu Gly Val Tyr Met His Ser Ser Gln145 150 155 160Trp Phe Glu Pro Phe Leu Phe Trp Phe Thr Val Lys Thr Glu Met Arg165 170 175Ser Val Ile Lys Ser Gln Thr His Thr Cys His Arg Arg Arg His Thr180 185 190Pro Val Phe Phe Thr Gly Ser Ser Val Glu Leu Leu Ile Ser Arg Asp195 200 205Leu Val Ala Ile Ile Ser Lys Glu Ser Gln His Val Tyr Tyr Leu Thr210 215 220Phe Glu Leu Val Leu Met Tyr Cys Asp Val Ile Glu Gly Arg Leu Met225 230 235 240Thr Glu Thr Ala Met Thr Ile Asp Ala Arg Tyr Thr Glu Leu Leu Gly245 250 255Arg Val Arg Tyr Met Trp Lys Leu Ile Asp Gly Phe Phe Pro Ala Leu260 265 270Gly Asn Pro Thr Tyr Gln Ile Val Ala Met Leu Glu Pro Leu Ser LeuSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16WO 98/13501 PCTIUS97/16718-136-275 280 285Ala Tyr Leu Gln Leu Arg Asp Ile Thr Val Glu Leu Arg Gly Ala Phe290 295 300Leu Asn His Cys Phe Thr Glu Ile His Asp Val Leu Asp Gln Asn Gly305 310 315 320Phe Ser Asp Glu Gly Thr Tyr His Glu Leu Ile Glu Ala Leu Asp Tyr325 330 335Ile Phe Ile Thr Asp Asp Ile His Leu Thr Gly Glu Ile Phe Ser Phe340 345 350Phe Arg Ser Phe Gly His Pro Arg Leu Glu Ala Val Thr Ala Ala Glu355 360 365Asn Val Arg Lys Tyr Met Asn Gln Pro Lys Val Ile Val Tyr Glu Thr370 375 380Leu Met Lys Gly His Ala Ile Phe Cys Gly Ile Ile Ile Asn Gly Tyr385 390 395 400Arg Asp Arg His Gly Gly Ser Trp Pro Pro Leu Thr Leu Pro Leu His405 410 415Ala Ala Asp Thr Ile Arg Asn Ala Gln Ala Ser Gly Glu Gly Leu Thr420 425 430His Glu Gln Cys Val Asp Asn Trp Lys Ser Phe Ala Gly Val Lys Phe435 440 445Gly Cys Phe Met Pro Leu Ser Leu Asp Ser Asp Leu Thr Met Tyr Leu450 455 460Lys Asp Lys Ala Leu Ala Ala Leu Gln Arg Glu Trp Asp Ser Val Tyr465 470 475 480Pro Lys Glu Phe Leu Arg Tyr Asp Pro Pro Lys Gly Thr Gly Ser Arg485 490 495Arg Leu Val Asn Val Phe Leu Asn Asp Ser Ser Phe Asp Pro Tyr Asp500 505 510Met Ile Met Tyr Val Val Ser Gly Ala Tyr Leu His Asp Pro Glu Phe515 520 525Asn Leu Ser Tyr Ser Leu Lys Glu Lys Glu Ile Lys Glu Thr Gly Arg530 535 540Leu Phe Ala Lys Met Thr Tyr Lys Met Arg Ala Cys Gln Val Ile Ala545 550 555 560SUBSTITUTE SHEET (RULE 26)?W0 98/1350]GluMetValValVal625AspSerTyrLeuLeu705ProGlyProLeuSer785ArgHisA311AlaSerLeu610ArgGlnAlaGluPro690TyrLeuGlyTyrVal770ThrAspLeuLeuLysGly595LysAlaAspPheThr675SerValCysIleLeu755GlnTrpTyrLysIleAsp580ValThrAlaThrIle660IlePheSerLysGlu740TyrGlyProPheAla820CASer565GluProHisLysAsp645ThrSerPheAspval725GlyLeuAspTyrVal805AsnSUBSTITUTE SHEET (RULE 26)AsnHisLysSerGly630HisThrLeuGlnPro710ProTyrAlaAsnAsn790IleGluGlyAspAspArg615PheProAspPheTrp695HisAsnCysAlaGln775LeuLeuThr- 137IleLeuLeu600SerIleGluLeuAla680LeuCysAspGlnTyr760ThrLysArgIleG12!Thr585LysProGlyAsnLys:665GlnProGlnLys745GluIleLysGlnVal82502265554 l999-03- 16Asn570LysGluValPheMet650LysArgLysProIle730LeuSerAlaArgArg810Serl’CT/US97/ 16718Tyr Phe Lys AspAlaSerHisPro635GluTyrLeuArgAsp715PheTrpGlyValGlu795LeuSerLeuHisThr620HisAlaCysAsnLeu700LeuIleThrValThr780AlaHisHisHisArg605SerValTyrLeuGlu685GluAspLysIleArg765LysAlaAspPheThr590GlyThrIleGluAsn670IleThrAlaTyrSer750IleArgArgIlePhe83 OAsn575LeuGlyLysArgThr655TrpTyrSerHisPro735ThrAlaValValGly‘e15ValGlyAlaP270A311Gln640ValArgGlyValVal720MetIleSerProThr800HisTyr?CA 02265554 1999-03-16W0 98/ 13501 PCT/US97/16718-138-Ser Lys Gly Ile Tyr Tyr Asp Gly Leu Leu Val Ser Gln Ser Leu Lys835 840 845Ser Ile Ala Arg Cys Val Phe Trp Ser Glu Thr Ile Val Asp Glu Thr850 855 860Arg Ala Ala Cys Ser Asn Ile Ala Thr Thr Met Ala Lys Ser Ile Glu865 870 875 880Arg Gly Tyr Asp Arg Tyr Leu Ala Tyr Ser Leu Asn Val Leu Lys Val885 890 895Ile Gln Gln Ile Leu Ile Ser Leu Gly Phe Thr Ile Asn Ser Thr Met900 905 910Thr Arg Asp Val Val Ile Pro Leu Leu Thr Asn Asn Asp Leu Leu Ile915 920 925Arg Met Ala Leu Leu Pro Ala Pro Ile Gly Gly Met Asn Tyr Leu Asn930 935 940Met Ser Arg Leu Phe Val Arg Asn Ile Gly Asp Pro Val Thr Ser Ser945 950 955 960Ile Ala Asp Leu Lys Arg Met Ile Leu Ser Ser Leu Met Pro Glu Glu965 970 975Thr Leu His Gln Val Met Thr Gln Gln Pro Gly Asp Ser Ser Phe Leu980 985 990Asp Trp Ala Ser Asp Pro Tyr Ser Ala Asn Leu Val Cys Val Gln Ser995 1000 1005Ile Thr Arg Leu Leu Lys Asn Ile Thr Ala Arg Phe Val Leu Ile His1010 1015 1020Ser Pro Asn Pro Met Leu Lys Gly Leu Phe His Asp Asp Ser Lys Glu1025 1030 1035 1040Glu Asp Glu Gly Leu Ala Ala Phe Leu Met Asp Arg His Ile Ile Val1045 1050 1055Pro Arg Ala Ala His Glu Ile Leu Asp His Ser Val Thr Gly Ala Arg1060 1065 1070Glu Ser Ile Ala Gly Met Leu Asp Thr Thr Lys Gly Leu Ile Arg Ala1075 1080 1085Ser Met Arg Lys Gly Gly Leu Thr Ser Arg Val Ile Thr Arg Leu Ser1090 1095 1100Asn Tyr Asp Tyr Glu Gln Phe Arg Ala Gly Met Val Leu Leu Thr GlySUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/13501 PCT/US97/ 16718-139-1105 1110 1115 1120Arg Lys Arg Asn Val Leu Ile Asp Lys Glu Ser Cys Ser Val Gln Leu1125 1130 1135Ala Arg Ala Leu Arg Ser His Met Trp Ala Arg Leu Ala Arg Gly Arg1140 1145 1150Pro Ile Tyr Gly Leu Glu Val Pro Asp Val Leu Glu Ser Met Arg Gly1155 1160 1165His Leu Ile Arg Arg His Glu Thr Cys Val Ile Cys Glu Cys Gly Ser1170 1175 1180Val Asn Tyr Gly Trp Phe Phe Val Pro Ser Gly Cys Gln Leu Asp Asp1185 1190 1195 1200Ile Asp Lys Glu Thr Ser Ser Leu Arg Val Pro Tyr Ile Gly Ser Thr1205 1210 1215Thr Asp Glu Arg Thr Asp Met Lys Leu Ala Phe Val Arg Ala Pro Ser1220 1225 1230Arg Ser Leu Arg Ser Ala Val Arg Ile Ala Thr Val Tyr Ser Trp Ala1235 1240 1245Tyr Gly Asp Asp Asp Ser Ser Trp Asn Glu Ala Trp Leu Leu Ala Arg1250 1255 1260Gln Arg Ala Asn Val Ser Leu Glu Glu Leu Arg Val Ile Thr Pro Ile1265 1270 1275 1280Ser Thr Ser Thr Asn Leu Ala His Arg Leu Arg Asp Arg Thr Thr Gln1285 1290 1295Val Lys Tyr Ser Gly Thr Ser Leu Val Arg Val Ala Arg Tyr Thr Thr1300 1305 1310Ile Ser Asn Asp Asn Leu Ser Phe Val Ile Ser Asp Lys Lys Val Asp1315 1320 1325Thr Asn Phe Ile Tyr Gln Gln Gly Met Leu Leu Gly Leu Gly Val Leu1330 1335 1340Glu Thr Leu Phe Arg Leu Glu Lys Asp Thr Gly Ser Ser Asn Thr Val1345 1350 1355 1360Leu His Leu His Val Glu Thr Asp Cys Cys Val Ile Pro Met Ile Asp1365 1370 1375His Pro Arg Ile Pro Ser Ser Arg Lys Leu Glu Leu Arg Ala Glu Leu1380 1385 1390SUBSTITUTE SHEET (RULE 25) ?CA 02265554 1999-03-16W0 98/13501 PCT/US97/ 16718- I140 -Cys Thr Asn Pro Leu Ile Tyr Asp Asn Ala Pro Leu Ile Asp Arg Asp1395 1400 1405Ala Thr Arg Leu Tyr Thr Gln Ser His Arg Arg His Leu Val Glu Phe1410 1415 1420Val Thr Trp Ser Thr Pro Gln Leu Tyr His Ile Leu Ala Lys Ser Thr1425 1430 1435 1440Ala Leu Ser Met Ile Asp Leu Val Thr Lys Phe Glu Lys Asp His Met1445 1450 1455Asn Glu Ile Ser Ala Leu Ile Gly Asp Asp Asp Ile Asn Ser Phe Ile1460 1465 1470Thr Glu Phe Leu Leu Ile Glu Pro Arg Leu Phe Thr Ile Tyr Leu Gly1475 1480 1485Gln Cys Ala Ala Ile Asn Trp Ala Phe Asp Val His Tyr His Arg Pro1490 1495 1500Ser Gly Lys Tyr Gln Met Gly Glu Leu Leu Ser Ser Phe Leu Ser Arg1505 1510 1515 ‘ 1520Met Ser Lys Gly Val Phe Lys Val Leu Val Asn Ala Leu Ser His Pro1525 1530 1535Lys Ile Tyr Lys Lys Phe Trp His Cys Gly Ile Ile Glu Pro Ile His1540 1545 1550Gly Pro Ser Leu Asp Ala Gln Asn Leu His Thr Thr Val Cys Asn Met1555 1560 1565Ile Tyr Thr Cys Tyr Met Thr Tyr Leu Asp Leu Leu Leu Asn Glu Glu1570 1575 1580Leu Glu Glu Phe Thr Phe Leu Leu Cys Glu Ser Asp Glu Asp Val Val1585 1590 1595 1600Pro Asp Arg Phe Asp Asn Ile Gln Ala Lys His Leu Cys Val Leu Ala1605 1610 1615Asp Leu Tyr Cys Gln Pro Gly Thr Cys Pro Pro Ile Arg Gly Leu Arg1620 1625 1630Pro Val Glu Lys Cys Ala Val Leu Thr Asp His Ile Lys Ala Glu Ala1635 1640 1645Arg Leu Ser Pro Ala Gly Ser Ser Trp Asn Ile Asn Pro Ile Ile Val1650 1655 1660SUBSTITUTE SHEET (RULE 26)?.................._............u..W... .CA 02265554 1999-03-16W0 98/13501 PCT/US97I16718-141-Asp His Tyr Ser Cys Ser Leu Thr Tyr Leu Arg Arg Gly Ser Ile Lys1665 1670 1675 1680Gln Ile Arg Leu Arg Val Asp Pro Gly Phe Ile Phe Asp Ala Leu Ala1685 1690 1695Glu Val Asn Val Ser Gln Pro Lys Ile Gly Ser Asn Asn Ile Ser Asn1700 1705 1710Met Ser Ile Lys Asp Phe Arg Pro Pro His Asp Asp Val Ala Lys Leu1715 1720 1725Leu Lys Asp Ile Asn Thr Ser Lys His Asn Leu Pro Ile Ser Gly Gly1730 1735 1740Asn Leu Ala Asn Tyr Glu Ile His Ala Phe Arg Arg Ile Gly Leu Asn1745 1750 1755 1760Ser Ser Ala Cys Tyr Lys Ala Val Glu Ile Ser Thr Leu Ile Arg Arg1765 1770 1775Cys Leu Glu Pro Gly Glu Asp Gly Leu Phe Leu Gly Glu Gly Ser Gly1780 1785 1790Ser Met Leu Ile Thr Tyr Lys Glu Ile Leu Lys Leu Asn Lys Cys Phe1795 1800 1805Tyr Asn Ser Gly Val Ser Ala Asn Ser Arg Ser Gly Gln Arg Glu Leu1810 1815 1820Ala Pro Tyr Pro Ser Glu Val Gly Leu Val Glu His Arg Met Gly Val1825 1830 1835 1840Gly Asn Ile Val Lys Val Leu Phe Asn Gly Arg Pro Glu Val Thr Trp1845 1850 1855Val Gly Ser Val Asp Cys Phe Asn Phe Ile Val Ser Asn Ile Pro Thr1860 1865 1870Ser Ser Val Gly Phe Ile His Ser Asp Ile Glu Thr Leu Pro Asn Lys1875 1880 1885Asp Thr Ile Glu Lys Leu Glu Glu Leu Ala Ala Ile Leu Ser Met Ala1890 1895 1900Leu Leu Leu Gly Lys Ile Gly Ser Ile Leu Val Ile Lys Leu Met Pro1905 1910 1915 1920Phe Ser Gly Asp Phe Val Gln Gly Phe Ile Ser Tyr Val Gly Ser Tyr1925 1930 1935Tyr Arg Glu Val Asn Leu Val Tyr Pro Arg Tyr Ser Asn Phe Ile SerSUBSTITUTE SHEET (RULE 26) ._........-—.m_.....m ~-— »- mm»?CA 02265554 1999-03-16W0 98/ 13501 PCT/US97/ 16718-142-1940 1945 1950Thr Glu Ser Tyr Leu Val Met Thr Asp Leu Lys Ala Asn Arg Leu Met1955 1960 1965Asn Pro Glu Lys Ile Lys Gln Gln Ile Ile Glu Ser Ser Val Arg Thr1970 1975 1980Ser Pro Gly Leu Ile Gly His Ile Leu Ser Ile Lys Gln Leu Ser Cys1985 1990 1995 2000Ile Gln Ala Ile Val Gly Asp Ala Val Ser Arg Gly Gly Ile Asn Pro2005 2010 2015Ile Leu Lys Lys Leu Thr Pro Ile Glu Gln Val Leu Ile Asn Cys Gly2020 2025 2030Leu Ala Ile Asn Gly Pro Lys Leu Cys Lys Glu Leu Ile His His Asp2035 2040 2045Val Ala Ser Gly Gln Asp Gly Leu Leu Asn Ser Ile Leu Ile Leu Tyr2050 2055 2060Arg Glu Leu Ala Arg Phe Lys Asp Asn Gln Arg Ser Gln Gln Gly Met2065 2070 2075 2080Phe His Ala Tyr Pro Val Leu Val Ser Ser Arg Gln Arg Glu Leu Ile2085 2090 2095Ser Arg Ile Thr Arg Lys Phe Trp Gly His Ile Leu Leu Tyr Ser Gly2100 2105 2110Asn Arg Lys Leu Ile Asn Arg Phe Ile Gln Asn Leu Lys Ser Gly Tyr2115 2120 2125Leu Ile Leu Asp Leu His Gln Asn Ile Phe Val Lys Asn Leu Ser Lys2130 2135 2140Ser Glu Lys Gln Ile Ile Met Thr Gly Gly Leu Lys Arg Glu Trp Val2145 2150 2155 2160Phe Lys Val Thr Ile Lys Glu Thr Lys Glu Trp Tyr Lys Leu Val Gly2165 2170 2175Tyr Ser Ala Leu Ile Lys Asp2180(2) INFORMATION FOR SEQ ID NO:7:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 15894 base pairs(B) TYPE: nucleic acidSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/13501-143-(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE:RNA (genomic)(xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:ACCAAACAAATCAAGATCCTTGAGGAGCTTGTGGAGCCATTTACCACTCGGCGGGCCCAAGTCAATTGATTTCAGAGTGAATGAGGCGGAGATGGTTCGATGATTCTGGGCAGACACGGCTAGTTGGTGAAGGACCTCTCGGAACAAACCGATTAGCCAGGACTGCATGAAAATGGGAGAGCGCAGGATCACTCCATGGGGGCAAGAGATGTTGGGTAAGATTATCAGGGAGCATTGTTCCAGAGGAATCATCCAGACTAACTAACAGGGTCAGAGGATCCCAGTCACAACCAATACTTTGAACAAGGAATACCATTCTAAGCTGATTCGATTTAGATTGTTTACGCCGATAGGATTGCTTTTTATCTTGATTTGCTGGTAACTGCACCCATACCCTCTGAGGTTTGAACGGTGAGGAGGGATAGATCAAACAAGAGCAGAAAAGAAACAAAACACATTACTGGACCGGTGCACTAATAGACCGATGACCTCTGGCCTTATCACATGATGATCTCAGATAGCCCAGATCTGAGCTAAGAAGAGAGAAAATTTCATGGTGGGAAATGATATACTATTAAGTGAGTTATCCATACATGGTAACTCTGGAGCTTTTGGTCGATTCAGCTGGAATCAATGATCAGATTAGGGATAGGACAAACCTTATAGTACCTGGTCAGGTTGTATATTATCCTGACGTTAGCCTTCGCATCATCCAAGCAGTTGAAGTGCAGGGTCTTGCTGGTGGATAAAGGTTGGATGTCTCTAATCCTGTGACATTGATTGGGATAGACACTTGAGTCTCCTAGAGAAATGCCATGGGCTTACTTTGAAGGTCAGTTCTATTCTAGTAATCCGAGATGACCCATTACAAATTCCTGGAAATTGGAAACCTTATTTGTGCATCAGGCTGAAGAGGTACCTAGTGATCAAAGATCCTGAGCGCAAAGGCGGTACACCCAAGGTGAGGAACGGATATCAAGTACATATATCAACTATGTATCTTGATGAATCTCAATTCAGAGTAGGAGTGTCCAGCATATCACATTGGCASUBSTITUTE SHEET (RULE 26)PCTIUS97/16718CACTTAGGAT 60GCCACACTTT 120TCAGGATCCG 180GATTCCTCAA 240CCGGATGTGA 300GAGTCTCCAG 360TTAGAGGTTG 420AACATGGAGG 480TCCAGGTCCG 540GGATTCAACA 600GTTACGGCCC 660CAAAGAAGGG 720AGGATTGCCG 780AGGACACCCG 840GTAGAGGCAG 900CCTGCTCTTG 960CTTTACCAGC 1020AACAAGTTCA 1080GAACTTGAAA 1140TTTAGATTAG 1200TCCGAACTCG 1260?W0 98/13501GTATCACTGCGGATCAGTAGGTGAGAATGAGAGAAGCCAGCCCATCCTCCCGCAGGACAGTGGAAGAACAATTAGGTGCGAAAACTTAGGGAGCCGATGGCTCAAGGCCGATATCAGACAGGTCTCAGCACGCGGTCAGGAATCTCCAGGGCGGTTAAGGAGCACCCTCTGATACCGAGGGCTTCTGATGAGAGGCAACAAGTAGGGCCATTTGGAACGGCCCTCGGAACGCCGCACTGAAATAATGAAGAAAACAGCCTCGAGGATGCAAGCGGTCGGAGCTACCAGGAGGAGAGCTACAACCAGCATGTCGAAGGTCAAGGCTCAGACAGAGGCCGAGAACCAGGTCCCAGAAGAGCAAGCCCATCGGATCCAGGACAAACCATGCTTGATCTGGAGACATCAAGCACGAATCCAAGACAGGAGGAGAGATATGCTATTTGAAACTGCACTTTCCGAAGCACTTCCGAAGATCGCGTCCGTCAGGGCCTACAGGAGTGAAGGGGGAGATGGCCAAAATCAAGGCTTGTTTCCCAGACAAGTTGGGGGGCAAGAGAAACCGCCCCTAGACAGCTGACGCTCACGGACACCCGACCAGAACAACACAGCCGCGGCACGCCATCTCACTGGCCGGACCGAGCCCTCAGCAATTAAGCGATGACTGGGTTACAGTGCTGACTCTCGATGAATCTCACTGACCGGAGAAGGAGGGGCTTGGGAAAGACACCCATTTTTATTGACAAGATGCACCTGACACCCGAACTATTATGATACACGAGGAT-144-CAGAGATTGCCCCAAGTATCAGGAAGACAGAGTCCAGCAGTTGACACTGCTGCTCAGGCTCTAGGGTATAACATCCGCCTCAGCCAACCAGTCAAAAACGGTCGAGGAAGGCCTGCAAGGGGATCAACTGGACGCTGAAATGTTATCATGATCATGGTTCGAAAACAGCGGGATCTGCTCGAGATCCACGACTCTCAATGAAAAAGGGGAGGTGGTGCAAGCGGGGAATGTCTGGTACCAGATGAGCTGTAATCAGAAGA02265554 1999-03-16AATGCATACTATTTCTACACGAGGGTCAAAAGCAAGTGATATCGGAGTCAGCAAGCCATGCAATGACAGAACCCTCCATCACCATCCACTGACTGGAATGCCATGGCAGCAAGAGGAGGCAAGGCGGTGCCTTTGGGAATTTTATGATCAAATCAGGCCTATGTGGATATCCATCTCTATAGCTCCTGAATTCCTCCGCCCAGACGCGAGCCCAATGTGCTCCCCGAGTGCAATCTCCCCTCTCCGATGTTAATCTCCAASUBSTITUTE SHEET (RULE 25)1‘PCT/US97l167l8ACTGAGGACA 1320GGTGATCAAA 1380CAGAGTCGGG 1440GCGAGAGCTG 1500GGCCAAGATC 1560GCAGGAATCT 1620GATCTTCTAG 1680ATTGTTATAA 1740CCCACGACTG 1800CATCCGGGCT 1860ATGGTCAGAA 1920AGGCAGTTCG 1980ACCTCGCATC 2040CCCCTCAAGA 2100CAGCGGTGAA 2160TGATGGTGAT 2220TGGCGAACCT 2280GGGGTTCAGG 2340ACTCCAATCC 2400CCCGAACCCC 2460ATTGGCCTCA 2520TCGAAAGTCA 2580TGTGAGCAAT 2640GAGATCCCAG 2700CCAAGACATC 2760GCTAGAATCA 2820?W0 98/13501TTGCTGTTATAGCATATCCAAAGGATCCCAGGCAGAGATTCTCCAGGGAACTAAAGCCGAGCATCACGCACGTTACCTGTCAGATGCTGACCAGTCGACCGCCTCCTAAGAAGGGTCGATTCAGAGTCATTGCTGGGGGTCCCTGCCCTTCTGAGCTTGAACAACACCCCTCAATGCAAATCCGTGTTGTGAAGAATGCTGGATTGACAACAACATTTATATTATTGCAAGCACCAGTCTGGTTCAAGAATCTGGAGGAGTGAAGGGAGACCCTGGAAGGACGACCCCACCAGGCCGAGCTGACTAATGGTCGGGAAAAAGTGTAATCCGTGACTCTCCTTGAAGATAATTAATTAGTACTTCCACAATGCGCTCCGATAAGATCCTGGTTGTTGAGGACAGGTGTTGGTCATAGTTGTTACTAACCCTCCCAAGTGTGCTTATATGAGCGGAATTCAGAGGCGATTGGCGGTCCACATCAATGAAAATCTCACATTAGAGACCTTATGTCAGATGCAAGCAAGTTGAGTCAACACCTCTCATGCAGATGTCACTGGCCGAAACGGACCAGTGGTGAGCTCACTCCATTATATGATGATATCAATGAAGTAGAACCTAAATCACAGAGATCTCAACCTACCACTAGGTGATAAGAGATCCCCAGATCCACAGAGACGTACAGCTCACACCTTAATGCGGTTAATCACCCGTCTCGGTCAATGCCTGGGAAGAGGGAACTTCAGAAAAGATGGAGCACAGGCATACCCACTGAATAGTAAGAA-145-ATTAAGAAGCAGCATCATGAGAACTCAATCGTTCTCAAGATCCAGAGGACGCCGTCGGGTAAATCCAGCCAAAGGAGCCACTACAGCTCACATTATAAAAACGACTTCGACCTACAGTGAGGAAGGATGATAGGGCCTCCCAAAACCCGACAGGGCTCAAGGAGAAAGGTATCTAATACCTTTCGGATAACAGTGGCCTTTCATCGACAAGGAGAAAGAAGCCTGGTTTTAAATGAGCAATGGATATCAATCCAGGCAGT02265554 1999-03-16AGATCAACAGTTGCCATTCCCCGACCTGAAAGCCCGTTGCAGCTGCTGAATTGTCCCTGAGGCTAGAGGAACGATCTTGCACTTACCTGCAACTTAGGAGCAAGTCGGCATGGCAGGCTGATGCTTTATGAATCGGGCGAGGAACTCCTCTGAAAAACTGCCTAACAACAGCTGGACACCCGGGTATTACCAACCTGCTATGCAGAGCAAGAGTGAAGTCTGCACTTGGTGACTCTCCATTGAAGACCTTTTTGCAGCCASUBSTITUTE SHEET (FIULE 26)PCTIUS97/16718GCAAAATATC 2880TGGACTTGGG 2940ACCCATCATA 3000CAGCCGACAA 3060GGAATTTCAA 3120CACCGGCCCT 3180GGATCGGAAG 3240CAAGTTCCAC 3300CAACCCCATG 3360CAAAGTGATT 3420TGGGACATCA 3480GTGCCCCAGG 3540TACATGTTTC 3600GCATTCGGGT 3660AAAGAGGCCA 3720GTGTTCTACA 3780GGGAGTGTCT 3840CCGCAGAGGT 3900ACCGTTCCCA 3960GTGACCCTCA 4020CTTCCTGAGG 4080TACTCTGCCG 4140GGGATAGGGG 4200GCACAACTCG 4260AATCGGTTAC 4320TCAGTTCCTC 4380?W0 98/ 13501AAGAATTCCGTGTAGACCGTGCCCGGACAAGCCGACAGCACCACCAGCCACGCTCCGGACACTGGAAGGCGACCGAGGTGACTAAACAAACGGCACCGCGCCCCGGTGCCAATCCAAGACGAGGAAGCCCGGGCCACCAGACCCCAGGCCTGGGGGACCCCTCCTCCTCTCCCACCCCTAGGTCTCAAGGACCGGTCAAAAGCTACAAAGATAACTCTCCACAGTTTTGGCAGAGTGTAGGCCCTAGGCGCTGAACTCTCCATTTACGACAGTGCCCAGCAAAAGCCCCCAGTGTGGACATCCCAATCCGACAGACCACCCCCTTCCCCCACCCAACCGCACTTAGGGCCCCCCCACCCCCACAGGTAGGGGGGGGCCCCACCCACCCCACTCCCAGACTCGATCCGGCGCCAAACCGCATCTCGAAGGGAAGGAGACACTGAATGTCTTTCCATTGGGGTTATGACTCGTCAATAACTGAACCAATTAGCTTCAAGTAGTTGCCACAGCAAGCCATCGACAGACGTGATCAAATACCCGAATCCAAAAGACCCAGGCGGCCCGTCCTCCTCAGCCGCATCCCTCCCCCAACAGGCATCCGAAAGGAACACACCGAAAACCACACACCAACCCCCCAAAAAACACACGACCACGGCCATCACGGAAGCCACCAAAGACATCAACCAAAAGATCGGGAATCCCTGCCATATTCCAATCTCTCTTTCCAGCCATCACGAGGGTAAGATGCACTTGAGACACAAGTGCTCAGATACAATCTGAGA-146-TAAATGATGAAACGACCCCCTTCACGGACCCAAGCACAGAGTAGGACCCCCCACAGCCCTGCAAGAACCCCTCCCTAGACCACACCCGACGAGGGAGCCCCCCGAACAGAAGGCCCCCAGCGGCAACCAACCCGAAAAAACAACCCGAACGTATCCCACCCAATCCACCAAGAATCAAGAATGGCAGTACAAGATAGGGGCAATCATTGGGAAATTGCAGAATGCAATGAAGATTTGCGGACAGCCGGCAGCAAGCCTGG02265554 1999-03-16CCAAGGACTACTCATAATGAAAGCGAGAGGACAGCCCCGACGAGGACCAACGGGAAAGGACACAACCGAAAGACCCTCCCAGAACCCAGACCAACCAATCCCCAGCACCCGGGCCGACAGACCAGAGCCCGGAAAGGCCACAGCACCCAAGCCTCTCCAACATCCGACGACTCATCCAATTGTTAACTCTTGGTAGGGATTCATAAAATTAATACAGGAGCCCAGAATATGAGTTGTCCTTTGCACTTCAAAACTACTAASUBSTITUTE SHEET (RULE 26)PCT/US97/ 16718TTCAAAGTTC 4440CAGCCAGAAG 4500CCAGCCAGCA 4560CACAAGGCCA 4620CCCCCAAGGT 4680ACCCCCAGCA 4740CCGCACAAGC 4800TCCCCGGCAT 4860CCCCGGCCCG 4920CCGCCGCCCC 4980AGCCACCGAC 5040CCAGCATCGC 5100AGACCACCCT 5160CAACCCGCGC 5220GAGCGATCCC 5280GTCCCCCGGT 5340CACTCAATTC 5400GTCCATCATG 5460CCAAACACCC 5520AGGAAGTGCA 5580AATGCCCAAT 5640ACTACTGAGA 5700AAGACCGGTT 5760GGCAGGTGCG 5820CCAGTCCATG 5880TCAGGCAATT 5940?W0 98/13501GAGGCAATCAATCAATAATGCTAGGGCTCACGGGACCCCAATCAATAAGGAGCAGAGGAAAGTATAGCCTGTCTCGTACACAAGGGTACCGTGTGCAGCCTCCACCAAGTTCACAAGGGAACGATCATTAGTGGTCGAGGTACCTGCACAAATCTGGGGACAGATATTGAGTGTGTCTTGAACAAAAAGGACATCAAAATCACAAGTCTCTGTCTCCGGCATCATCCACATCCTAAGGGATTTGCTGGCTCATTAGACTCGGCAAGCAGGAGCTGATACCAATTGCTCAGTATCTGCGGATATTAGAAAATAAAGGCCCGATCCGACGCTATATAGGCTCTTATCTCGAAAAAATGCCTTCCTGTGCTCGACCTAATAGCATCAAGACCCTGAACGGTGTGAATTGACCTATGCAATTGCGGAGTATGAAGAGGGTTGATGGGAACAAGTCCTATGTAAGCTCTCCGTCATTCCCTCTGGATGTCACCACAGTAGGATAGGTTCTATTCGCATCGGGCAGCAGCAGGAGATGGTCTATGAACATACTATACAGATATCCATCGCTCGGATACGATAACTCACGTCCGAGATTTCAAGAGTGGTTTTGATGAGGTACCCGATGTACACTCGTACAATTGTGCATGACAAGATCGACCATCCAACGGTCCTCCCTAAGCTGGAGAGGTTTATCGAGGGATCCCCTGGTATGTCAGTCGCTCTGATCAAGCAACCCCGAACGATAACCGAGACCGTTATTAACAGTCATGTTTCTCCATCTACAC-147-ATATTGGCTGCAACTATCTTGAAATCCTGTCAGGCTTTGAAGTGGAGGTGGTCGACJLCAGAAGGGGGTGATATACCACTGTCATCGTGTAAGTCCTCTGCTCCGGGTCTTTCAATCCTCTCTAACATACAGTCGGGAGCAATATCATTGGGATGCCAAGGAGCACTAGCAGCTTTAATATAGACCAGGCCTCCCCTACAAACCGCATCCATCGGTAGTTAAATAAATGCCAGAACATCTTGAGCTTGATCCGCAGAGATC02265554 1999-03-16TTCAGGGTGTGTGATTTAATCATTATTTGGGCTATGCGCTATTTACTGGGAGTCCTACTTTTGTCCACCGTGCCCAAGTACTTTCATGCCTCCAAGAATGTTGGGAACCGGCAAGTGTTATTGCTGCCGAGGAGGTATCCAGAAGTTGGAAATTGCTGGATAGTTTACATGTTGCTGCAGTAAAGCCTGACTCTTGAAACGCATCAAGGCATTAAAACTTTTCTACAAAGATGATTGATAGGGTTGCTAGCATAAGAGCCSUBSTITUTE SHEET (RULE 26)PCT/US97/16718CCAAGACTAC 6000CGGCCAGAAG 6060CCCCAGCTTA 6120TGGGGGAGAT 6180CATCTTAGAG 6240CATTGTCCTC 6300GCTAGAGGGG 6360TGTTGCAACC 6420AGAGGGGACT 6480CCTCCGGGGG 6540GTTCATTTTA 6600CACAACAGGA 6660TCACTGCCCG 6720GGACGCGGTG 6780CGTAGGGACA 6840GTCATCGGAC 6900CCTGATTGCA 6960GGGGCGTTGT 7020TCTTACAGGG 7080ACAGATTTCC 7140CACCCGAAAT 7200AGGGTGCAAG 7260ACAACCCCCA 7320GACCTTATGT 7380CCATTGCAGG 7440TCAGCACCAA 7500?W0 98/13501TCTAGATGTAGATCATCGGTCATCTCTGACTTGGTGTATCGGCTGCTGAACAATCAGTTCATTCTCAAACATCTATAGTCTAATCTGAGCAGGGGTTATCGCAACCAGTCAGCCCTCTGTCAGCTTCCAGCCCCCTATCATATCGCTGACAATGGAGACACGAGTGGGCAGAGTCTGACACGGTTCAGGGGCCAATGAAGGGTTAGTCCCAACATACCTAACCTGGTCAGTGTGGTTTATGCCTATAAAGCTGGTGCCGTACTAACTCAAGATGAAGTGGAAAATTAAATAACCCGCCAGGAACTCATGACTAGCTGTCTATGTCGCTGTACCATGACATAGTAAAGGGTAGAAATCCGGAGTAATGATTCACAGGGAAGCTCGTCAAGCACGGATGATCAATCAAGCAATGCTTCCAGCCCATTGAAGGGTTGAGCTTAATGGACCTATAACCTAGCCTAACCTCTTCACCTGCGGAGGGATCTCCAATTATGTTTACAGGGGTCCCAACACTTCTGTGCATCGAGCATCAGCCTGAGGACTCCTTAATCCAGAGAATCAAATGCATTGGTCAAAGGGAAACCCTGTTGGACCCAGGGAATCAGAGTTGTCGTTTGGGGGCTCAGCAACTGATTCTGTCACTAGGTGTCTGCAGTGATAGAAATGGGCTGTAGGCGTGTAAATAACAGGATAAATCAAAATACAAAACCAATAGGTGTAATCTGTTCCAATTGGATGGTGAATGTTTTGGCGCCCAGGCCGTCGAATTACATGCTTGCGGA-148-GGTCAAGGACACCTCAGAGAGGATAGGGAGATTGGATTATGAACTCAACTCTGCTCAGGGCTTGTATTTAGTACGGGGGAACAACTGAGCTCCGGTGTTCCATGGTGGCTGGTTCCCTATGAAATCCCCATAGGCTTTACCCCGACAACAGGGTAAAAACTCCTTCATACTGCTTCAGGACCACAACAATCAACACATTGCAAGGAAGCATGTCAAACTCAACCTACGATCTCATTTTCTAGTGGAATGCTTCAGAATCT02265554 1999-03-16GTGCTGACACTTCACTGACCTACGACTTCAGATCAATACTCTACTGGAGGCCCACTACAAAGTCGAGGTTACTTACCTAGATGCACCGAGCATATGACAATTGGGGGAGCCAGGGGTCAGACCGACATGCCTCTCATCTCCGGACAGATGCAAGCACTCTGGGGTCTTGTTTCGGGCCATGTGTATTGGCGAGTGGATACGGCGAGGACTAGTTCCAATCACTTCCAGGGTACTTTTATCTTCACATGGGGGTGGACATASUBSTITUTE SHEET (RULE 26)PCT/US97Il 6718CACTCTTCAA 7560T§GTGAAATT 76201GAGATCTCAC 7680GTGCAGATGT 7740CCAGGGCAAC 7800TCAGAGGTCA 7860ACAATGTGTC 7920TGGGAAAGCC 7980TGTTTGAAGT 8040ACTATTTTGA 8100TCAGGTTCGC 8160GGAAAGGTGT 8220AATCCTGGGT 8280ACAGAGGTGT 8340ACAAGTTGCG 8400GCGAGAATCC 8460CTGTTAATCT 8520TGATCACACA 8580TGACTATCCC 8640CGAGATTCAA 8700GCCATGCCCC 8760TGGTAATTCT 8820TTGAACATGC 8880CTTTTAGGTT 8940ACCAAAAACT 9000TCACTCACTC 9060?...........-—._..... ......... ,.W0 98/13501TGGGATGGTGATAGGGCTGCGTGAAATAGACGCTATCTGTATAAGATAGTCTACACTGTGTAAACAATGTCTCATATTCCCAAGGAAGATAGGTTTTCCAAGGACATCAAAGCCCTTTCTCCCATACTTGTGTTAATCTCTGACGTTTGACCGCTATGACAACTGATAGATGGAGCCACTCTTTCCTTAAATGAAGGTACTACATCTGACCAGTAACGGCAGACTCTGATGGCACGGAGGATGCTCAAGCTTGCTGGAGTGGCATGGGAGCAGTGAACCGCATCAGAATTCAACCAGATCAGCTATCCTGTCAGAACATCGGAAGTTGGGATATCCAAATCCGTGAGCTCATGCCTGAGGGGAGAAAATTGTTTTGGTTTCCATAGGAGGTCGTGACCTTACTGGTTTTGCATTGATGCTTGGTTTCTTCTTCACTTGCTCCACTGCTTTTTATCATGAGAGGGGAGATTTGCTGAAAATGAAGGGTCATCAGTTGGCCATTCAGGTGAAGAGATTTGGCCATCAGCTGCACATCACATGATAAGAAAAACGTTGTACCCTGGAGTATGCTCAAGCACCGCCAATGTCATCATGTAATCAGGCTAAAAAAGGGACACTAACTATTAACTTGGACAGTCAAGAAGACACACACGTTGCTATAAATGTATTGTGAGGTATGCAGCCTGCACTCGTACCTGCAACACTGAAATACTTAATTGAAGTTCTCATTTTGTCAGGAAATGCCATATTTTCCCCTGACCCGGGTTAACACTGTTTTATGC-149-AGTCACTCGGGTCACTCAGATAGGGTCCAAAAGTTCACCTGAGTCCCTCATAAAAAACGGAGTCCAAGCTATTTATTTAAGAAATTCGCTCACGGCTTGGGAGTTTACATCTGAGATGAGCTGTATTCTTTCAGTAAGGAATGTCATAGAAACTTCTAGGGGAATCCAACTGAGGGACATATGATGTTCTCCTTAGATTATCAGAAGTTTACATGAATCAGTGGAATCITTCCCCCTGCAATGAGCAGTGCTCTTAGCCT02265554 1999-03-16GAAGATGGAACACCAGGCATGTGGTTTCCCAGATAGCCCGCGCTTACAGCATTCTCCAACTAGGAGTTATCATAGAAGACGTACTCCAAACCTAGGCTCCGCACAGCTCCGTCAGTGATTCACTGGTAGTGTCTCAACATGGGGAGGTTAAAGAGTCAGATTATCAAATTAACAGTAGAATGACCAAAACCATTTTCATACGGCCACCCCGCCTAAAGTCAATCAACGGCTGCTGCAGACCGTTGATAACGGACAGTGATSUBSTITUTE SHEET (IRULE 26)_..._._._........—--u-—-.—....._...... .PCT/US97/16718CCAATCGCAG 9120ACCCACTAGT 9180GTCATGGACT 9240ATAGTTACCA 9300CTTGAGGACC 9360CAAATGATTA 9420CCGGCCCACT 9480AAAGAGTCAA 9540GTCAGTGATA 9600GAATTGAGGG 9660CAATGGTTTG 9720AAATCACAAA 9780TCAGTTGAGC 9840GTATATTACC 9900ATGACAGAGA 9960TACATGTGGA 10020GTAGCTATGC 10080CTCAGAGGTG 10140GGGTTTTCTG 10200ACTGATGACA 10260AGACTTGAAG 10320ATTGTGTATG 10380TATCGTGACA 10440ACAATCCGGA 10500TGGAGATCAT 10560CTGACAATGT 10620?W0 98/13501ACCTAAAGGAAGTTCCTGCGTTAATGATTCTCCATGACCCGTAGACTTTTTAATCTCAAAATTTGACTAAGTCACAGGGGGGAACGTTAAACACTGATCAATCTCAAGAATAAATGAGATCTGTCCTCTAGCAAAGTCCCGTCAGAAGCTGGGTAAGGATTACCCAGCACACTTTGTAATCAATTGTTTCTGTCCCAATCAAACAAGGGCATGACCGTTACTCTTGGCTTACAACGATCTTGAACATGAGATCTCAAGAGCAAGGCACTTTTACGATCCTGAGCTTTGACTGAGTTCAATCGCTAAAATGCGGGATTGGCGGCACTCCACGGGGCCAGTCAGCAGAAAAATCCGGAGAATGTACTGCCTTTTACGGATTATGTAAGTGATCAATGACCAAGTGGACCATCTGCCTCGTTAATGGCCTTACTCTTAGGCAAATCACATTTTACTCAAGAGCAGCATGCAGTTCTTGCATATCACAATCAATCTTAATAAGGCAGGCTGTTTAATGATTCTCCAGCTGCTCTCCCCCAAGGGAACCATATGATACTGTCTTACAACTTACAAAAAAGTATTTTAACTCTGGCTGTTAAAAACCTGGGTTTGTAGATAGAAACCTAATTGGAGATCCCTCATTTTCCTCATTGCCATCTTCATCAAGCACCATTCGTGCAAGGGGAACCTTAAGAAGGCTACATGTTTGTCTATTATTGCAAGATAATATTGCTATCCCTGAACGTCAACCATGAATGGCACTGTGTCAGAAACAGCATCACTAA-150-AAAGGGAATGCCGGGTCACGTGATAATGTAGCCTGAAAGATGAGGGCATGAGGACAATGGTCTCAGGAGTACTCCCGAAGGATTCCCTCAACGAGACAGTATGAGACCATTTCAGTGGCTCCCCCGACCTAGTACCCTATCCTACTTATAACAATCAGACAACGGGAAGCACATTGGCCACAAAAGGAATGTGTATTCTGCAACAATGGCTCCTAAAAGTCCCGAGATGTTGCCCGCTCCTCGGTGATCCTGCCTGAAGA02265554 1999-03-16GGATTCAGTTGAGGCTTGTATGTCGTAAGTAAAGGAGATCCCAAGTGATCGATGGCCAAGCCCCAAAGATCCCAGTCCACTGTAATTCGGCAGCGCATTTCAGCTTATTTGCATAAGAGGTGACGCCCATGGGAGGTATACCTGGCTGCTCATAGCCGTATGCTAGAGTATCACCTCAAGATATTATGATGTCAGAGACTTAAAAGCATCGATACAGCAAAGTCATACCCTATTGGGGGGAGTAACATCAGACCCTCCATSUBSTITUTE SHEET (RULE 26)PCT/US97l16718TACCCGAAAG 10680GATGTTTTCC 10740GGAGCCTACC 10800AAGGAAACAG 10860GCTGAAAATC 10920GATGAGCACG 10980CTCAAAGAAA 11040ACAAGTACCA 11100CAGAATCAAG 11160ATCACGACTG 11220GCACAGAGGC 11280CTTGAAACCT 11340GTCCCGTTAT 11400GAAGGGTATT 11460TATGAGAGCG 11520ACAAAAAGGG 11580ACTAGAGATT 11640GCAAATGAGA 11700GGGCTACTTG 11760ATAGTTGATG 11820GAGAGAGGTT 11880ATTTTGATCT 11940CTCCTCACAA 12000ATGAATTATC 12060TCAATTGCTG 12120CAAGTAATGA 12180?W0 98/13501CACAACAACCTTGTATGCGTTCCATAGTCCAGAGACTGGCTCCTGGATCAAAGGCCTGATTGTCCAATTAGAAATGTCCTATATGTGGGCTAGAATCTATGATCAGTCAAAGGAAACATCTGAAGCTTGCCAGTGTACTCCAAGGCAAAGCGACTAATTTCCCTTGTCAGCAGATAAGAATTTTAGAAACTTCACGTCGACCCGCAAGCTCTTTAATTGAAATTTGTTACCTATGATTGATAGGGGATGATCACCATCTAGGGGGACTCTCCAGAGCATCAAACCCAATGGGCATTCCTCTAGTGTCACATCGAGCCAGCTGACTATGAACATTGACAAAAAGACTAGCTGCGAGGCCACCTACGGATGGATCCTTGAGACTTCGTAAGAATGGGCTTACGGCCAATGTGAGCGCATAGGAGTGGCAAGGAGTTGATACTATTGTTTCGAAACAGATTGTAGAGCTGAGGCAGAGATGCAATGGTCCACACCTGGTAACACGATATCAATCTTGGGCCAGCATCATTCCTAGACTAGACTCCTTAAAAGGGTATGGACAGGCGGGGCAAGAGATGAGGAAGGCAATTTAGAGGAGTCATGTTCGAGGACGGCCTTATTCGGCTTTTTTGTCCGTCCCATATAGCCCCAAGTAGGTGATGATGAGCCTGGAGGTTGAGGGATCTATACCACAAAACTTTATATCTCGAGAAAGTGCGTGATCCGCAGAGCTATACAAGGCTATCCCCAACTATAAATTTGAGAAGTTTCATAATGTGCAGCCA-151-ACTGGGCTAGTCAAGAACATTATTCCATGAATATTATAGTAGTCTATTGCGGGGGTTAACCAGGGATGGTCAGTGCAGCTCTATTTACGGGTCATGAGACCCTCGGGTTGTTGGTTCTACGATCCTTGCGATAGCTCTTGAGCTAAGGGTGTAGCACTCATCTCCAACGAACCAACAAGGATACTGGATCCGATGATAGAGTACCAACCCACACCCAGAGATCACATTCTAGGACCATATCTGAGTTTCTTCAATTG$GC02265554 1999-03-16CGACCCTTACAACTGCAAGGTGACAGTAAAACCTAGGGCAAGGCATGCTACTCTCGAGTGGCTATTGACAGGCTAGAGCCCCTTGAGGTCATGTGTCATCCCAACTGGATCACTGATGAGATCTGCCGTTGAACGAAGCCGATCACTCCCAGTGAAATACCAATCTCTCAAATGCTTCTAATCTAACACGTCATCCCAGGATTGATATATCCATAGGAGGAGCTAAGTCCGAATGAAATTGCTTATAGAGATTTGATGTASUBSTITUTE SHEET (RULE 26)PCT/US97/16718TCAGCAAATC 12240TTTGTCCTAA 12300GAAGAGGACG 12360GCTCATGAAA 12420GATACCACAA 12480ATAACCAGAT 12540GGAAGAAAGA 12600CTAAGAAGCC 12660CCTGATGTAC 12720TGCGAGTGTG 12780GATATTGACA 12340AGAACAGACA 12900AGAATAGCAA 12960TGGTTGTTGG 13020ATCTCGACTT 13080TCAGGTACAT 13140TTTGTCATAT 13200GGGTTGGGTG 13260GTATTACATC 13320ATACCCAGCT 13380GATAATGCAC 13440CACCTTGTGG 13500ACAGCACTAT 13560TCAGCTCTCA 13620CCAAGATTAT 13680CATTATCATA 13740?W0 98/13501GACCATCAGGAAGGAGTGTTGGCATTGTGGCAACTGTGTGAAGAGTTAGAGATTCGACAAGGACCTGCCCATATCAAGGCTTGTAGACCAGATTGKGAGTCAAAGGTCGGATGATGTTGCGGGGTAGTCTCTTGCTACAAACGGC1TGTTAACT##HCAAAATTAGCACCTTGTCAAGGTTCAATTTCATAGACCTTACCTGGCTCTACTGGGATTTTGTTCTACCCTAGAAGCTAACCGGGACTTCACCCAATTGTGGGGAAATATCAGTAAGGTGCTTTATTATAGAGCAACATGGTTAGAGTTCACACATCCAGGCAACCGATTCGAAGAGGCTAGGTTACTCATGCTGATCCAGGACAGCAACAACAAAATTGCTC"rGccAA'r'rA'rAGCTGTTGAGCTTGGGTGAGGTGCTTCTATCTATCCCTCCGCTCTTTAACAGTCAGTAATCAACAAAGATCCTTGGCAAATCAGGGATTTGTACAGCAACGCTAATGAATTGGACTTATAAGGCGCAGTTCAATGGGTGAGCGTCAATGCTCCCTATCCATGTACACATGCTTTTCTTTTGTAAACACTTGTGGTCTAAGGCTTATCTCCAGTCTCTGACTTTTCATTTTTGATCTCAAATAAAAGATATCAGAAATCCATGATATCAACATGGGTCGGGTTAATAGTGGGGGAAGTTGGCCGGGAGGCCCGATCCCTACCTACTATAGAGAATAGGATCAAATAAGCTATGTTCATATCTACCTGAAAAGAGGTCACATCCAGTAGAGGTG-152-TGTTGTCTTCTAAGCCACCCGTCCTTCACTATATGACCTAGTGAAAGCGAGTGTTCTGGCCGGTAGAGAACAGGATCTTCATCTCCGTCGATGCCCTCGCTGAGCATCAAACACAAGCAACTTTCCGCAGTAATTAGGAGCTATGTTGATTTTCCGCCAATTGTCGAACAAAGTCACGTGCTAGTGTGGGAGTTAGAGGATACTGGTGATTAGGGTCTCACTGAATCTTATTAAGCAGCATATCTATCAAATATCAACCC02265554 1999-03-16GTTCCTTTCTAAAGATCTACTGATGCTCAACCTTGACCTGTGAGGATGTAAGATTTGTACATGTGCAGTTGTGGAACATAAGGATCTATCTGAGGTAAATGGATTTCAGAGCACAATCTTAATCGGGTTAATGCCTTGAGCACTTATAAGTTCTAGATCTCAGAATGGGAGGTAGGCAGTATTTATCCATATTGGCAGCCTAAGCTTATGTTATAGAGAATTTAGTTATGGATAATTGAAGCAACTAAGCTATTCTGAAASUBSTITUTE SHEET (RULE 26)PCT/US97/16718AGAATGAGCA 13800AAGAAATTCT 13860AACTTGCACA 13920TTGTTGAATG 13980GTACCGGACA 14040TGTCAACCAG 14100CTAACCGATC 14160AATCCAATTA 14220AAACAGATAA 14280GTCAGTCAGC 14340CCTCCACACG 14400CCCATTTCAG 14460AACTCATCTG 14520CCAGGGGAAG 14580GAGATACTAA 14640GGTCAAAGGG 14700GTAGGTAATA 14760ATAGATTGCT 14820TCAGATATAG 14880ATCTTATCGA 14940CCTTTCAGCG 15000GTGAACCTTG 15060ACAGATCTCA 15120TCATCTGTGC 15180TGCATACAAG 15240AAACTTACAC 15300?CA 02265554 1999-03-16W0 98/13501 PCT/US97/16718- 153 -CTATAGAGCA GGTGCTGATC AGTTGCGGGT TGGCAATTAA CGGACCTAAG CTGTGCAAAG 15360AATTAATCCA CCATGATGTT GCCTCAGGGC AAGATGGATT GCTTAACTCT ATACTCATCC 15420TCTACAGGGA GTTGGCAAGA TTCAAAGACA ACCAAAGAAG TCAACAAGGG ATGTTCCACG 15480CTTACCCCGT ATTGGTAAGT AGTAGGCAAC GAGAACTTGT ATCTAGGATC ACTCGCAAAT 15540TTTGGGGGCA TATTCTTCTT TACTCCGGGA ACAGAAAGTT GATAAATCGG TTTATCCAGA 15600ATCTCAAGTC CGGTTATCTA ATACTAGACT TACACCAGAA TATCTTCGTT AAGAATCTAT 15660CCAAGTCAGA GAAACAGATT ATTATGACGG GGGGTTTAAA ACGTGAGTGG GTTTTTAAGG 15720TAACAGTCAA GGAGACCAAA GAATGGTATA AGTTAGTCGG ATACAGCGCT CTGATTAAGG 15780ATTAATTGGT TGAACTCCGG AACCCTAATC CTACCCTAGG TAGTTAGGCA TTATTTGCAA 15840TATATTAAAG AAAACTTTGA AAATACGAAG TTTCTATTCC CAGCTTTGTC TGGT 15894(2) INFORMATION FOR SEQ ID NO:8:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 2183 amino acids(B) TYPE: amino acid(C) STRANDEDNESS:(D) TOPOLOGY: linear(ii) MOLECULE TYPE: protein(xi) SEQUENCE DESCRIPTION: SEQ ID NOx8:Met Asp Ser Leu Ser Val Asn Gln Ile Leu Tyr Pro Glu Val His Leu1 5 10 15Asp Ser Pro Ile Val Thr Asn Lys Ile Val Ala Ile Leu Glu Tyr Ala20 25 30Arg Val Pro His Ala Tyr Ser Leu Glu Asp Pro Thr Leu Cys Gln Asn35 40 45Ile Lys His Arg Leu Lys Asn Gly Phe Ser Asn Gln Met Ile Ile Asn50 55 60Asn Val Glu Val Gly Asn Val Ile Lys Ser Lys Leu Arg Ser Tyr Pro65 70 75 80Ala His Ser His Ile Pro Tyr Pro Asn Cys Asn Gln Asp Leu Phe Asn85 90 95SUBSTITUTE SHEET (‘RULE 26)?ICA 02265554 1999-03-16WO 98/13501 PCT/US97/ 16718-154-Ile Glu Asp Lys Glu Ser Thr Arg Lys Ile Arg Glu Leu Leu Lys Lys100 105 110Gly Asn Ser Leu Tyr Ser Lys Val Ser Asp Lys Val Phe Gln Cys Leu115 120 125Arg Asp Thr Asn Ser Arg Leu Gly Leu Gly Ser Glu Leu Arg Glu Asp130 135 140Ile Lys Glu Lys Ile Ile Asn Leu Gly Val Tyr Met His Ser Ser Gln145 150 155 160Trp Phe Glu Pro Phe Leu Phe Trp Phe Thr Val Lys Thr Glu Met Arg165 170 175Ser Val Ile Lys Ser Gln Thr His Thr Cys His Arg Arg Arg His Thr180 185 190Pro Val Phe Phe Thr Gly Ser Ser Val Glu Leu Leu Ile Ser Arg Asp195 200 205Leu Val Ala Ile Ile Ser Lys Glu Ser Gln His Val Tyr Tyr Leu Thr210 215 220Phe Glu Leu Val Leu Met Tyr Cys Asp Val Ile Glu Gly Arg Leu Met225 230 235 240Thr Glu Thr Ala Met Thr Ile Asp Ala Arg Tyr Ala Glu Leu Leu Gly245 250 255Arg Val Arg Tyr Met Trp Lys Leu Ile Asp Gly Phe Phe Pro Ala Leu260 265 270Gly Asn Pro Thr Tyr Gln Ile Val Ala Met Leu Glu Pro Leu Ser Leu275 280 285Ala Tyr Leu Gln Leu Arg Asp Ile Thr Val Glu Leu Arg Gly Ala Phe290 295 300Leu Asn His Cys Phe Thr Glu Ile His Asp Val Leu Asp Gln Asn Gly305 310 315 320Phe Ser Asp Glu Gly Thr Tyr His Glu Leu Ile Glu Ala Leu Asp Tyr325 330 335Ile Phe Ile Thr Asp Asp Ile His Leu Thr Gly Glu Ile Phe Ser Phe340 345 350Phe Arg Ser Phe Gly His Pro Arg Leu Glu Ala Val Thr Ala Ala Glu355 360 365SUBSTITUTE SHEET (RULE 26)?W0 98/1350]AsnLeu385ArgAlaHisGlyLys465ProArgMetAsnLeu545GluMetValValVal625AsnVal370MetAspAlaGluCys450AspLysLeuIleLeu530PheAsnAlaSerLeu610LysGlnArgLysArgAspGln435PheLysGluValMet515SerAlaLeuLysGly595LysAlaAspLysGlyHisThr420qrsMetAlaPheAsp500TyrTy!‘LysIleAsp580ValThrGluThrTyrHisGly405IleValProLeuLeu485ValValSerMetSer565GluProTyrLysAspSUBSTITUTE SHEET (RULE 26)CAMetAla390GlyArgAspLeuAla470ArgPheValLeuThr550AsnHisLysSerGly630HisAsn375IleSerAsnA811Ser455AlaTyrLeuSerLys535TyrGlyAspAspArg615PhePro- 155GlnPheTrpAlaTrp440LeuLeuAspA811Gly520GluLysIleLeuLeu600SerValGluPIGCysProGln425ArgAspGlnPIOAsp505AlaLysMetGlyThr585LysProGlyA811LysGlyPIO410AlaSerSerArgPro490SerTyrGluArgLys570LysGluValPheIle02265554 l999-03- 16ValIle395LeuSerPheAspGlu475LysSerLeuIleAla555TyrAlaSerHisPro635GluIle380IleThrGlyAlaLeu460TrpGlyPheHisLys540CysPheLeuHisThr620HisThrValIleLeuGluGly445ThrAspThrAspAsp525GluGlnLysHisArg605SerValTyrTyrAsnProGly430ValMetSerGlyPro510ProThrValAspThr590GlyThrIleGluPCT/US97/16718GluGlyLeu415LeuArgTyrValSer495TyrGluGlyIleAsn575LeuGlyArgArgThrThrTyr400HisThrPheLeuTyr480ArgAspPheArgAla560GlyAlaP150AsnGln640Val?ICA 02265554 1999-03-16W0 98/13501 PCT/US97/16718-156-645 650 655Ser Ala Phe Ile Thr Thr Asp Leu Lys Lys Tyr Cys Leu Asn Trp Arg660 665 670Tyr Glu Thr Ile Ser Leu Phe Ala Gln Arg Leu Asn Glu Ile Tyr Gly675 680 685Leu Pro Ser Phe Phe Gln Trp Leu His Lys Arg Leu Glu Thr Ser Val690 695 700Leu Tyr Val Ser Asp Pro His Cys Pro.Pro Asp Leu Asp Ala His Val705 710 715 720Pro Leu Cys Lys Val Pro Asn Asp Gln Ile Phe Ile Lys Tyr Pro Met725 730 735Gly Gly Ile Glu Gly Tyr Cys Gln Lys Leu Trp Thr Ile Ser Thr Ile740 745 750Pro Tyr Leu Tyr Leu Ala Ala Tyr Glu Ser Gly Val Arg Ile Ala Ser755 760 765Leu Val Gln Gly Asp Asn Gln Thr Ile Ala Val Thr Lys Arg Val Pro770 775 780Ser Thr Trp Pro Tyr Asn Leu Lys Lys Arg Glu Ala Ala Arg Val Thr785 *790 795 800Arg Asp Tyr Phe Val Ile Leu Arg Gln Arg Leu His Asp Ile Gly His805 810 815His Leu Lys Ala Asn Glu Thr Ile Val Ser Ser His Phe Phe Val Tyr820 825 830Ser Lys Gly Ile Tyr Tyr Asp Gly Leu Leu Val Ser Gln Ser Leu Lys835 840 845Ser Ile Ala Arg Cys Val Phe Trp Ser Glu Thr Ile Val Asp Glu Thr850 855 860Arg Ala Ala Cys Ser Asn Ile Ala Thr Thr Met Ala Lys Ser Ile Glu865 870 875 880Arg Gly Tyr Asp Arg Tyr Leu Ala Tyr Ser Leu Asn Val Leu Lys Val885 890 895Ile Gln Gln Ile Leu Ile Ser Leu Gly Phe Thr Ile Asn Ser Thr Met900 905 910Thr Arg Asp Val Val Ile Pro Leu Leu Thr Asn Asn Asp Leu Leu Ile915 920 925SUBSTITUTE SHEET (RULE 26)?W0 98/1350]Met Ala930ArgMet Ser945ArgIle Ala AspThr Leu HisAla995Asp TrpIle Thr Arg1010Ser Pro Asn1025Glu Asp GluPro Arg AlaLeuLeuLeuGln980SerLeuP130ArgCA— 157Ala935Leu Pro Pro IleVal950Phe Arg Asn IleLys Met Ile Leu965ArgThr Gln Gln985Val MetSer Ala1000Asp Pro TyrAsn Ile Thr1015Leu LysMet Leu Lys Gly Leu1030Leu Ala Ala Phe Leu104502265554 1999-03-16PCT/U S97! 16718Gly Gly Met Asn Tyr940Leu A311Gly Pro Val Thr Ser960Asp Ser955Ala970Glu Glu975Ser Leu Met ProPro Gly Asp Ser Ser Phe Leu990Asn Leu Val Cys Val Gln1005SerAla Phe Val Leu1020Arg Ile HisPhe Glu1040His Asp Asp Ser1035LyeMet Asp Arg His Ile1050Ile Val1055Ala His Glu Ile Leu Asp His Ser Val Thr Gly Ala Arg106010651070Glu Ser Ile Ala Gly Met Leu Asp Thr Thr Lys Gly Leu Ile Arg Ala10751080Ser Met Arg Lys Gly Gly Leu Thr Ser10901095Asn Tyr Asp Tyr Glu Gln Phe Arg Ala11051110Arg Lys Arg Asn Val Leu Ile Asp LynAla Arg Ala Leu Arg Ser11401125His Met Trp11451085Arg Val Ile Thr Arg Leu Ser1100Gly Met Val Leu Leu Thr Gly1115 1120Glu Ser Cys Ser Val Gln Leu1130 1135Ala Arg Leu Ala Arg Gly Arg1150Pro Ile Tyr Gly Leu Glu Val Pro Asp Val Leu Glu Ser Met Arg Gly1155His Leu Ile Arg Arg His11701160Glu Thr Cys11751165Val Ile Cys Glu Cys Gly Ser1180Val Asn Tyr Gly Trp Phe Phe Val Pro Ser Gly Cys Gln Leu Asp Asp118511901195 1200SUBSTITUTE SHEET (RULE 26)?ICA 02265554 1999-03-16WO 98113501 PCT/US97/16718-158-Ile Asp Lys Glu Thr Ser Ser Leu Arg Val Pro Tyr Ile Gly Ser Thr1205 1210 1215Thr Asp Glu Arg Thr Asp Met Lys Leu Ala Phe Val Arg Ala Pro Ser1220 1225 1230Arg Ser Leu Arg Ser Ala Val Arg Ile Ala Thr Val Tyr Ser Trp Ala1235 1240 1245Tyr Gly Asp Asp Asp Ser Ser Trp Asn Glu Ala Trp Leu Leu Ala Arg1250 1255 1260Gln Arg Ala Asn Val Ser Leu Glu Glu Leu Arg Val Ile Thr Pro Ile1265 1270 1275 1280Ser Thr Ser Thr Asn Leu Ala His Arg Leu Arg Asp Arg Ser Thr Gln1285 1290 1295Val Lys Tyr Ser Gly Thr Ser Leu Val Arg Val Ala Arg Tyr Thr Thr1300 1305 1310Ile Ser Asn Asp Asn Leu Ser Phe Val Ile Ser Asp Lys Lys Val Asp1315 1320 1325Thr Asn Phe Ile Tyr Gln Gln Gly Met Leu Leu Gly Leu Gly Val Leu1330 1335 1340Glu Thr Leu Phe Arg Leu Glu Lys Asp Thr Gly Ser Ser Asn Thr Val1345 1350 1355 1360Leu His Leu His Val Glu Thr Asp Cys Cys Val Ile Pro Met Ile Asp1365 1370 1375His Pro Arg Ile Pro Ser Ser Arg Lys Leu Glu Leu Arg Ala Glu Leu1380 1385 1390Cys Thr Asn Pro Leu Ile Tyr Asp Asn Ala Pro Leu Ile Asp Arg Asp1395 1400 1405Ala Thr Arg Leu Tyr Thr Gln Ser His Arg Arg His Leu Val Glu Phe1410 1415 1420Val Thr Trp Ser Thr Pro Gln Leu Tyr His Ile Leu Ala Lys Ser Thr1425 1430 1435 1440Ala Leu Ser Met Ile Asp Leu Val Thr Lys Phe Glu Lys Asp His Met1445 1450 1455Asn Glu Ile Ser Ala Leu Ile Gly Asp Asp Asp Ile Asn Ser Phe Ile1460 1465 1470Thr Glu Phe Leu Leu Ile Glu Pro Arg Leu Phe Thr Ile Tyr Leu GlySUBSTITUTE SHEET (RULE 26)?W0 98/ 13501CA 02265554 1999-03-16-159-1475 1480 1485Gln Cys Ala Ala Ile Asn Trp Ala Phe Asp Val His Tyr His Arg Pro1490 1495 1500Ser Gly Lys Tyr Gln Met Gly Glu Leu Leu Ser Ser Phe Leu Ser Arg1505 1510 1515 1520Met Ser Lys Gly Val Phe Lys Val Leu Val Asn Ala Leu Ser His Pro1525 1530 1535Lys Ile Tyr Lys Lys Phe Trp His Cys Gly Ile Ile Glu Pro Ile His1540 1545 1550Gly Pro Ser Leu Asp Ala Gln Asn Leu His Thr Thr Val Cys Asn Met1555 1560 1565Val Tyr Thr Cys Tyr Met Thr Tyr Leu Asp Leu Leu Leu Asn Glu Glu1570 1575 1580Leu Glu Glu Phe Thr Phe Leu Leu Cys Glu Ser Asp Glu Asp Val Val1585 1590 1595 1600Pro Asp Arg Phe Asp Asn Ile Gln Ala Lys His Leu Cys Val Leu Ala1605 1610 1615Asp Leu Tyr Cys Gln Pro Gly Thr Cys Pro Pro Ile Arg Gly Leu Arg1620 1625 1630Pro Val Glu Lys Cys Ala Val Leu Thr Asp His Ile Lys Ala Glu Ala1635 1640 1645Arg Leu Ser Pro Ala Gly Ser Ser Trp Asn Ile Asn Pro Ile Ile Val1650 1655 1660Asp His Tyr Ser Cys Ser Leu Thr Tyr Leu Arg Arg Gly Ser Ile Lys1665 1670 1675 1680Gln Ile Arg Leu Arg Val Asp Pro Gly Phe Ile Phe Asp Ala Leu Ala1685 1690 1695Glu Val Asn Val Ser Gln Pro Lys Val Gly Ser Asn Asn Ile Ser Asn1700 1705 1710Met Ser Ile Lys Asp Phe Arg Pro Pro His Asp Asp Val Ala Lys Leu1715 1720 1725Leu Lys Asp Ile Asn Thr Ser Lys His Asn Leu Pro Ile Ser Gly Gly1730 1735 1740Ser Leu Ala Asn Tyr Glu Ile His Ala Phe Arg Arg Ile Gly Leu Asn1745 1750 1755 1760SUBSTITUTE SHEET (RULE 26)PCTlUS97l167l8?CA 02265554 1999-03-16W0 98/13501-160-Ser Ser Ala Cys Tyr Lys1765 1770Cys1780 1785Ser Met Leu Ile Thr Tyr1795 1800Tyr Asn Ser Gly Val Ser1810 1815 1820Ala Pro Tyr1825 1830 1835Gly Asn Ile Val Lys Val Leu Phe Asn1845Gly Arg Pro Glu1850Val Gly Ser1860 1865Ser Ser Val Gly Phe Ile His Ser Asp Ile Glu Thr Leu1875 1880Asp Thr Ile Glu Lys Leu Glu Glu Leu Ala1890 1895Ala Ile Leu1900Ile Leu Val Ile1915Leu Leu Leu Gly Lys Ile Gly Ser1905 1910LysPhe Ser Gly Asp Phe Val Gln Gly Phe Ile Ser Tyr Val1925 1930Tyr Arg Glu Val Asn Leu Val Tyr Asn1940Pro Arg Tyr Ser1945Thr Glu Ser Tyr Leu Val Met Thr Asp Leu Lys Ala1955 1960AsnAsn Pro Glu Lys Ile Lys Gln Gln Ile Ile Glu Ser Ser1970 1975 1980Ser Pro Gly Leu Ile Gly His Ile Leu Ser Ile Lys Gln1985 1990 1995Ile Gln Ala Ile Val Gly Gly Ala Val Ser Arg Gly Asp2005 2010Ile Leu Lys Lys Leu Thr Pro Ile Glu Gln Val Leu Ile2020 2025SUBSTITUTE SHEET (RULE 26)18851965PCT/US97/16718Ala Val Glu Ile Ser Thr Leu Ile Arg Arg1775Leu Glu Pro Gly Glu Asp Gly Leu Phe Leu Gly Glu Gly Ser Gly1790Lys Glu Ile Leu Lys Leu Asn Lys Cys Phe1805Ala Asn Ser Arg Ser Gly Gln Arg Glu LeuPro Ser Glu Val Gly Leu Val Glu His Arg Met Gly Val1840Val Thr Trp1855Ile Asp Cys Phe Asn Phe Ile Val Ser Asn Ile Pro Thr1870Pro Asn LysSer Met AlaLeu Met Pro1920Gly Ser His1935Phe Ile Ser1950Arg Leu MetVal Arg ThrLeu Ser Cys2000Ile Asn Pro2015Se: Cys Gly2030?CA 02265554 1999-03-16WO 98/13501-161-Leu Ala Ila Aen Gly Pro Lye Leu Cye2035 2040Lys Glu Leu Ile Hie His2045Val Ala Ser Gly Gln Aep Gly Leu Leu Aen Ser Ile Leu Ile Leu2050 2055 2060Arg Glu Leu Ala Arg Phe Lye Asp Aen2065 2070Gln Arg Ser Gln Gln Gly2075Phe Hie Ala Tyr Pro Val Leu Val Ser2085Ser Arg Gln Arg Glu Leu2090Ser Arg Ile Thr Arg Lye Phe Trp Gly His Ile Leu Leu Tyr Ser2100 2105 2110Aen Arg Lye Leu Ile Aen Arg Phe Ile Gln Aen Leu Lye Ser Gly2115 2120 2125Leu Ile Leu Asp Leu Hie Gln Aen Ile Phe Val Lye Aen Leu Ser2130 2135 2140Ser Glu Lye Gln Ile Ile Met Thr Gly Gly Leu Lye Arg Glu2145 2150 2155Phe Lye Val Thr Val Lye Glu Thr Lye Glu Trp Tyr Lye Leu2165 2170 2175Tyr Ser Ala Leu Ile Lye Asp2180(2) INFORMATION FOR SEQ ID NO:9:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 15894 base pairs(B) TYPE: nucleic acid(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: RNA (genomic)(xi) SEQUENCE DESCRIPTION: SEQ ID N039:ACCAAACAAA GTTGGGTAAG GATAGTTCAA TCAATGATCA TCTTCTAGTG CACTTAGGATTCAAGATCCT ATTATCAGGG ACAAGAGCAG GATTAGGGAT ATCCGAGATG GCCACACTTTTAAGGAGCTT AGCATTGTTC AAAAGAAACA AGGACAAACC ACCCATTACA TCAGGATCCGGTGGAGCCAT CAGAGGAATC AAACACATTA TTATAGTACC AATCCCTGGA GATTCCTCAAsuesrrrurs sneer (RULE 25)2095PCT/US97/16718AspTyrMet2080ValGlyTyrLyeTrp Val2160Val Gly60120180240?W0 98/13501TTACCACTCGGCGGGCCCAAGTCAATTGATTCCAGAGTGAATGAGGCGGAGATGGTTCGATGATTCTGGGCAGACACGGCTGGTTGGTGAAGGACCTCTCGAAACAAACCGATTAGCCAGGACTGCATGAAAATGGGGGAGTGCAGGATCACTCCATGGGGGCAAGAGATGTATCACTGCAGATCAGTAGGTGAGAATGAGAGAAGCCAGCCCATCTTCCCGCAGGACAGCGGAAGAACAACTAGGTGCGAAAACTTAGGATCCAGACTTACTAACAGGGTCAGAGGATCCCAGTCACAACCAATACTTTGAACAAGGAATACCATCCTAAGCTGATTCGATTTAGATTGCTTACGCCGACAGGATTGCTTTTTATCCTGATTTGCTGGTAACTGCACCCATACCCTCTGAGGTTTGAACGGTAAGGAGGCGAGGATGCAAGCGGTTGGAGCTACCGAGAGGAGAGCTACAACCGGCACATCGAAGGTCAAGGCTCAGACAGAGGCCGAGAACCAGGTCCCACTGGACCGGTGCACTAATAGACCGATGACCTCTGGCCTTATCACATGATGATCTCAGATAGCCCAAATTTGAGCTAAGAAGAGAGAAAATTTCATGGTCGGAAATGATATACTATTAAGTGAGTTATCCATACATGGTAACTCTGGAGCTTTTGGCCGATTCAGCTGGAAAGGCTTGTTTCCCAGACAAGTTGGGGGGCAAGAGAAACCGCCCCTAGACAGCTGACGCCCACGGACACCCGGCCAGAACAACACAGCCGC-162-TGGTCAGGTTGTATATTATCCTGACGTTAGCCTTCGCATCATCCAATTAGTTGAAGTGCAGGGTCTTGCTGGTGGATAAAGGTTGGATGTCTCTAATCCTGTGACATTGATTGGGATAGACACTTGAGTCTCCTGGAGAAATGCCATGGGCTTACTTTGAAGGTCAGTTCCAGAGATTGCCCCAAGTATCAGGAAGATAGGGCCCAGCAGTTGACACTGCTGCTTAGGCTCTATAGTGTAACATCCGCCTCAGCCCATCA02265554 1999-03-16AATTGGAAACCTTATTTGTGCATAAGGCTGAAGAGGTACCTAGTGATCAAAGACCCTGAGCGCAAAGGCGGTACACCCAAGGTGAGGAACGGATATCAAGTACATATATCAACTATGTATCTTGATGAACCTCAATTCAGAGTAGGAGTGTCCAGCATATCACATTGGCAAATGCATACTATTTCTACACGAGGGTCAAAAGCAAGTGATAACGGAGTCCGCAAGCCATGCAATGACAGAACCCTCCATCACCATCCACTSUBSTITUTE SHEET (RULE 26)PCT/US97/16718CCGGATGTGA 300GAGTCTCCAG 360TTAGAGGTTG 420AACATGGAGG 480TCCAGGTTCG 540GGATTCAACA 600GTTACGGCCC 660CAAAGAAGGG 720AGGATTGCCG 780AGAACACCCG 840GTAGAGGCAG 900CCTGCTCTTG 960CTTTACCAGC 1020AACAAGTTCA 1080GAACTTGAAA 1140TTTAGATTAG 1200TCTGAACTCG 1260ACTGAGGACA 1320GGTGATCAAA 1380CAGAGTCGAG 1440GCGAGAGCTG 1500AGCCAAGATC 1560GCAGGAATCT 1620AATCTTCTAG 1680ATTGTTATAA 1740CCCACGATTG 1800?W0 98l1350lGAGCCAATGGCTCAAGGCCGATATCAGACAGGTCTCAGCACGCGGTCAGGAATCTCCAGGGCGGTTAAGGAGCACCCTCTGATACCGAGGGCTTCTGATGAGAGGCAACAGGTAGGGCCATTTGGAACGGCCCTCGGAACGCCGCACTGAAATAATGAAGAAAACAGCCTCTGCTGTTATAGCATATCCAAAGGATCCCAGGCAGAGATTCTCCAAGGAACTAAAGCCGAGCATCACGCACGTTACCTGACAGATGCTGACAGAAGAGCAAGCCCATCGGACCCAGGACAAACCATGCCTGACCTGGAGACATCAAGCACGAATCCAAGACAGGAGGAGAGATATGCTATTTGAAACTGCACTTTCCGAAGCACTTCCGGAGATCGCGTCCATCAGGGCCTACAGGAGTGAAGGGGGAGATGGCCAAAATTGAAGGGAGACCCTGGAAGGACGACCCCACCAGGCCGAGCTGACAAATGGTCGGGAAAAAGTGTAATCCGTGACTCTCCTTGAAGATAATCAGGCACGCCATCTCACTGGCCGGAGCGAGCCCTCAGCAATTGAGCGATGACTGGGTTACAGTGCTGACTCTCAATGAATCTCACTGACCGGAGAAGGAGGGGCTTGGGAAAGACACCCATTTTTATTGACAAGGTGCACCTGACACCCGAACTATTATGATACACGAGGATAGTTGAGTCAACACCTCTCATGCAGATGTCACTGGCCGAAACGGACCAGTGATGAGCTCACTCCATTATATGATGATATCAATGAAGTAG-163-GTCAAAAACGATCGAGGAAGACCTGCAGGGGGATCAACTGGACGCTGAAATGTTATTACGATCATGGTTCGAAAACAGCGGGATCTGCTCGAGATCCACGACTCTCAATGAAAAAGGGCAGGTGGTGCAAGCGGGGAATGTCTGGTACCAGATGAGCTGTAATCAGAAGAATTAAGAAGCAGCATCATGAGAAATCAATCGTTCTCAAGATCCAGAGGACGCCGTCGGGTAAATCCAGCCAAAGGAGCCACTACAGCTCA02265554 1999-03-16GACTGGAATGCTATGGCAGCAAGAGAAGGCAAGGCGGTGCCTTTGGGAATTTTATGATCAAATCAGGCCTATGTGGATATCCATCTCTATAGCTCCTGAGTTCCTCCGCCCAGACGCGAGCCCAATGTGCTCCCCGAGTGCAATCTCCCCTCTCTGATGTTAATCTCCAAAGATCAACAGTCGCCATTCCCCGACTTGAAAACCCGTTGCAGCTGCTGAATTGTTCCTGAGGCTAGAGGAATGATCTTGCACTTACCTGCSUBSTITUTE SHEET (RULE 25)PCT/US97/16718CATCCGGGCT 1860ATGGTCAGAA 1920AGGCAGTTCG 1980ACCTCGCATC 2040CCCCCCAAGA 2100CAGCGGTGAA 2160TGATGGTGAT 2220TGGCGAACCT 2280GGGGTTCAGG 2340ACTCCAATCC 2400CCCGGACCCC 2460ATTAGCCTCA 2520TCGAAAGTCA 2580TGTGAGCAAT 2640GAGATCCCAG 2700CCAAGATATT 2760GCTAGAATCA 2820GCAAAATATC 2880TGGACTTGGG 2940ACCCATCATA 3000CAGCCGACAA 3060GGAATTTCAG 3120CACCGGCCCT 3180GGATCGGAAG 3240CAAGTTCCAC 3300CAACCCCATG 3360?W0 98/13501CCAGTCGACCGCCTCCCAAGAAGGGTCGATTCAGAGTCATTGCTGGGGGTCCCTGCCCTTCTGAGCTTGAACAACACCCCTCAACGCAAATCCGTGTTGTGAAGAATGCTGGATTGACAACAACATTTATATTATTGCAAGCACCAGTCTGGTTCAAGAATCTGGAGGAGAAGAATTCCGTGTAGACCGTGCCCGGACAAGCCGACGGCACCACCAGCCATGCCCCCGATATTGGAAGGCGACCGAGGTGACTAAACAAACAACTAGTACTTCCACAATGCGCTCCGATAAGATCCTGGTTGTTGAGGACAGGTGTTGGCCATAGTTGTTACTAACTCTCCCAAGTGTGCTTATATGAGCGGAATTCAGAGGCGATAGGCGGTCCACATCAATGAAAATCTCACATTAGAGACCTTATGTCAGATGCAAGCATTTACGACAGTGCCCAGCAAAAGCCCCCAGCGCGAACACCCCAATCTGCCAAACCACCCCCTCCCCCTACCCAACCGCACTTAGGGCCCAAACCTAAATCACAGAGACCTCAACCCACCACTAGGCGACAAGCGATTCCCAGATCCACAGAGACGTACAGCTCACACCTTAATGCGGTTAATCACCCGTCTCGGTCAATGCCTGGGAAGAGGGAACTTCAGAAAAGATGGAGCACAGGCATACCCGCTGAATAGTAAGAAGACGTGATCAAATGCCCGAATCCGAAAGACCCAGGCGGCCCATCCTCCTCAACCGCATCCCTTCCTCAACAGGCATCCGAAAGGAACATA-164-CATTATAAAAACGACTTCGACCTACAGTGAGGAAGGATGATAGGGCCTCCCAAAGCCCGACAGGGCTCAAGGAGAAAGGTATCTGATACCTTTCGGATAACAGTGGCCTTTCATCGACAAGGAGAAAGAAGCCTGGTTTTAAATGAGCAATGGATATCAATCCAGGCAGTTAAATGATGAAACGACCCCCTCCACGGACCCCAGCACAGAGTGGGACCCCCCACCACCCCACAAGAACTCCTCCCTAGACCACACCCAAC02265554 1999-03-16AACTTAGGAGCAAGTCGGCATGGCAGGCTGATGCTTTATGAATCGGGCGAAAAACTCCTCTGAAAAACTGCCTAACAACAGCTCGATACCCGGGTATTACCAACCTGCTGTACAGAGCAAGAGTGAAGTCTGCACTTGGTGACTCTCCATTGAAGACCTTTTTGCAGCCACCAAGGACTACTCACAATGAAAGCGAGAGGACAGCCCCGACGAGGACCAACGGGAAAGAACACAACCGAAAGATCCTCTCAGAACCCAGASUBSTITUTE SHEET (RULE 26)PCT/US97/16718CAAAGTGATT 3420TGGGACATCA 3480GTGCCCCAGG 3540TACATGTTTC 3600GCATTTGGGT 3660AAAGAGGCCA 3720GTGTTCTACA 3780GGGAGTGTCT 3840CCGCAGAGGT 3900ACCGTTCCTA 3960GTGACCCTTA 4020CTTCCTGAGG 4080TACTCTGCCG 4140GGGATAGGGG 4200GCACAACTCG 4260AATCGATTAC 4320TCAGTTCCTC 4380TTCAAAGTTC 4440CAGCCAGAAG 4500CCAGCCAGCA 4560CACAAGGCCA 4620CCCCCAAGGC 4680ACCCCCAGCA 4740CCGCACAAGC 4800TCCCCGGCAA 4860CCCCGGCCCA 4920?W0 98/ 13501CGGCGCCGCGCCCCGGTGCCAATCCAAGACGAGGAAGCCCGGGCCACCAGACCCCAGCCCCGAAGGACCCCTCCTCCTCTCCCACCCCTAGGTCTCAAGGACCGGTCAAAAGCTACAAAGATAACTCTCCACAGTTTTGGCAGAGTGTAGGCCCTAGGCGCTGAACTCTCGAGGCAATCAATCAATAATGCTCGGGCTCACGGGACCCCAATCAATAAGGAGCGGAGGAAAGTATAGCCTGTCTCGTACACAAGGGTACCCCCCCAACCCCACAGGCAGGGGGGGGGCCCACCCACCCCACTCCCAGACTCGATCCGGCGCCGAACCGCATCTCGAAGGGAAGGAGACACTGAACGTCTCTCCATTGGGGTTATGACTCGTCAATAACTGAACCAATTAGCTTCAAGTAGTTGCCACAGCAAGCCATCGAGACAAGCAGGAGCTGATACCAATTGCTCAGTATCTGCGGATGTTAGAAAATAAAGGCCCGATCCGACGCTACATAGGCTCTTATCTCGAACACCGACAACCAGACACCAACCCCCCAAAAAACACACGACCACGGCCATCACGGGAGCCACCAAGGACACCAACCAAAAGATCGGGAATCCCTGCCATATTCCAATCTCTCTTTCCAGCCATCACGAGGGTAAGATGCACTTGAGACACAAGTGCTCAGATACAATCTGAGAGCAGGAGATGGTCTATGAACATACTATACAGATATCTATCGCTCGGATACGATAACTCACGTCCGAGATTTCAAGAGTGGTTTTGATGAG-165-GAGGGAGCCCCCCGAACAGAAGGCCCCCAGCGGCAACCAACCCGCAGAAACAACCCGAACGTATCCCACACAATCCACCAAGAATCAAGAATGGCAGTACAAGATAGGGGCAATCATTAGGAGATTGCAGAATGCAATGAAGATTTGCGGACAGCCGGCAGCGAGCCTGGATATTGGGTGCAACTATCTTGAAATCCTGTCAGGCTTTGAAGTGGAGGTGGTCGACACAGAAGGGGGTGATATACCACTGTCATCGTGTA02265554 1999-03-16CCAACCAATCCCCAGCACCCGGGCCGACAGACCAGAACCCGGAAAGGCCACAGCACCCAAGCCTCTCCAACACCCGACGACTCATCCAATTGTTAACTCTTGGTAGGAATTCATAAAATTAATACAGGAGCCCAGAATATGAGTAGTCCTTTGCACTTCAAAACTACTAATTCAGGGTGTGTGATTTAATCATTATTTGGGCTATGCGCTATTTACTGGGAGTCCTACTTTTGTCCACCGTGCCCAAGTACTTTCATGCCSUBSTITUTE SHEET (IRULE 26)PCT/US97/16718CCGCCGGCTC 4980AACCATCGAC 5040CCAGCACCGC 5100AGACCACCCT 5160CAACCCGCGC 5220GAGCGATCCC 5280GTCCCCCGGT 5340CACTCAACTC 5400GTCCATCATG 5460CCAAACACCC 5520AGGAAGTGCA 5580AATGCCCAAT 5640ACTACTGAGA 5700AAGACCGGTT 5760GGCAGGTGCG 5820CCAGTCCATG 5880TCAGGCAATT 5940CCAAGACTAC 6000CGGCCAGAAG 6060CCCCAGTTTA 6120TGGAGGAGAC 6180CATCTTAGAG 6240CATTGTCCTC 6300GCTAGAGGGG 6360TGTTGCAACC 6420AGAGGGGACT 6480?W0 98/13501GTGTGCAGCCTACACCAAGTTCACAAGGGAACGATCATTAGTAGTCGAGGTACTTGCACAAATCTGGGGACAGATATTGAGTGTGTCTTGAACAAAAAGGACATCAAAATCACAAGTCTC_TATCTCCGGCATCATCCACATCCCAAGGGATTTGCTGGCTCATTAGACTTTCTAGATGTAAATCATCGGTCATCTCTGACTTGGTGTATCGGCTGCTGAACAATCAGTTCATTCTCAAACATCTATAGTCTAATCTGAGCAAAATGCCTTCCTGTGCTCGACCTAATAGCATCAAGACCCTGAACGGCGTGAATTGACCTATGCAATTGCGGAGTATGAAGAGGGTTGATGAGAACAAGTCCTATGTAAGCTCTTCGTCATTCCCTCTGGATGTCACCACAGTAGGATAGGTTCTGTTTGCATCGGGCAGACTAACTCAAGATGAAGTGGAAGATTAAATAACCCGCCAGGAGCTCATGACTAGCTGTCTATGTCGCTGTACTATGACATAGCAAAAGGTCAGTACCCGATGTACACTCGTACAATTGTGCATGACAAGATCGATCATCCAACGGTCCTCCCTAAGTTGGAGAGGTTTATCGAGGGATCCCCTGGTATGTCAGTCGCTCTGATCAAGCAACCCCGAACAATAAACGAGACCGTCATTAACAGTCATGTTTCTCCATCTACACTCGAGCATCAGCCTGAGGACTCCTTAATCCAGAGAATCAAATGCATTGGTCAAAGGGAAACCCTGTTAGACCCAGGGAATCAGAGTTGTC-166-AGTCCTCTGCTCCGGGTCTTTCAATCCTTTCTAACATACAGTCGGGAGCAATATCATTGGGATGCCAAGGAGCACTAGCAGCTTTAATATAGACCAGGCCTCCTCTACAAACCGCACCCATCGGTAGTTAGATAAATGCCAGAACATCTTGAGCTTGATCCGCAGAGATCGGTCAAGGACACCTCAGAGAGGATAGGGAGATTGGATTATGAACTCAACTCTGCTCAGGGCTTGTATTTAGTATGGGGGAACAACTGAGC02265554 1999-03-16TCCAAGAATGTTGGGAACCGGCAAGTGTTATTGCTGCCGAGGAGGTATCCAGAGGTTGGAAATTGTTGGATAGTCTACATGTTGCTGCAGTAAAGCCTGACTCTTGAAACGCATCAAGCCATTAAAACTTTTCTACAAAGATGATTGATAGGGTTGCTAGCATAAAAGCCGTGCTGACACTTCACTGACCTACGACTTCAGATCAATACTCTACTGGAGACCCACTACAAGGTCGAGGTTACTTACCTAGATGTACCGAGSUBSHTUTESHEET(RULE26)PCTIUS97/16718ccrccssscs 6540GTTCATTTTA 6600CACAACAGGA 6660TCACTGCCCG 6720AGACGCTGTG 6780CGTAGGGACA 6840GTCATCGGAC 6900CCTGATTGCA 6960GGGGCGTTGT 7020TCTTACGGGA 7080ACAAATGTCC 7140CACCTGAAAT 7200AGGGTGCAAG 7260ATAACCCCCA 7320GACCTTATGT 7380CCATTGCAGG 7440TCAGCACCAA 7500CACTCTTCAA 7560TAGTGAAATT 7620GAGATCTCAC 7680GTGCAGATGT 7740CCAGAACAAC 7800TCAGAGGTCA 7860ACAATGTGTC 7920TGGAAAAGCC 7980TGTTTGAAGT 8040?W0 98/ 13501AGGTGTTATCGCAACCAGTCAGCCCTTTGTCAGCTTCCAGCCCCTTATCATATCGCTGACAATGGAGACACGAGTGGGCAGAGTCTGACACGGTTCAGGGGCCAATGAAGGGTTAGTCCCAACATACCTAACCTGGTCAATGTGGTTTATGCCTATAAAGCTGGTGCCGTTGGGATGGTGATAGGGCTGCGTGAAATAGACGCTATCTGTATAAGATAGTCTACACTGTGTAAACAATGTCTCATATTCCCGAGGAAGATAGAAATCCGGAGTAATGATCCACGGGGAAGCTCGTCAAGCACGGATGATCAATCAAGCAATGCTTCCAACCCATTGAAGGGTTGAGCTTAATGGACCTATAACCTAGCCTTACCTCTTCACCTGCGGAGGGATCTCCAATTACGTTTACAGGGGTCCCCACACTTCTGTGGGCATGGGAGTAGTGAACCACATCAGAATTCAACCAGATCAGCCATCCTGTCAGAACATCGGAAGTTGGGATATCCAAATCCGTGAACTCCAGTTTGGGGGCTCAGCAACTGATTCTATCACTAGGTGTCTGCAGTGATAGAAATGGGCTGTAGGCGTGTAAATAACAGGATAAATCAAAATACAAATCCAATAGGTGTAATCTGTCCCAATTGGATGGTGAATGTTTTGGCGCCCAAGCCGTCGAATTACATGCTTGCGGATCAGCTGCACATCACATGATAAGAAAAACGTTATACCCTGGAGTATGCTCAAGCACCGCCAATGTCATCATGTAATCAGGCTCAAAAAGG-167-TCCGGTGTTCTATGGTGGCTAATTCCCTATGAAATCCCCACAGGCTTTACCCCGACAACAGGGTAAAATCTCCTTCATACTGCTTCGGGACCACAACAATCAACACATTGTAAGGAAGCATGTCAAACTCAACCTACGATCTCATTTTCTAGTGGAATGCCTCAGAATCTAGTCACCCGGGTCACCCAGATAGGGTCCAAAAGTTCACCTGAGTCCCTCATAAAAAACGGAGTCCAAGCTATTTATTTAAGGAATTCGCT02265554 1999-03-16CATATGACAATTGGGGGAGCCAGGGATCAGACCGACATGCCTCTCATCTCCGAACAGATGCAAGCACTCTGGGGTCTTGTTTCGGGCCATGTGTATTGGCGAGTGGATACGGCGAAGACTAGTTCCAATCACTTCCAGGGTACTTTTATCTTCACATGGGGGTGGACATAGAAGATGGAACATCAGGCATGTGGTTCCCCAGATAGCCCGCGCTTACAGCATTTTCCAACTAGGAGTTATCATAGAAGACGTACTCCAAASUBSTITUTE SHEET (RULE 26)PCT/US97/16718ACTATCTTGA 8100TCAAACTCGC 8160GGAAAGGTGT 8220AATCCTGGGT B280ACAGAGGTGT 8340ACAAGTTGCG 8400GCGAGAATCC 8460CTGTTGATCT 8520TGATCACACA 8580TGACTATCCC 8640CGAGATTCAA 8700GCCATGCCCC 8760TGGTGATTCT 8820TTGAACATGC 8880CTTTTAGGTT 8940ACCAAAAACT 9000TCACTCACTC 9060CCAATCGCAG 9120ACCCACTAGT 9180GTTATGGACT 9240ATAGTTACCA 9300CTGGAGGACC 9360CAAATGATTA 9420CCGGCCCACT 9480AAAGAGTCAA 9540GTCAGTGATA 9600?WO 98/13501AGGTTTTCCAAGGACATCAAAGCCCTTTCTCCCATACTTGTGCTAATCTCTGACATTTGACCGCTATGACAACTGATAGATGGAGCCTCTCTTTCCTTAAATGAAGGTACTACATCTGACCAGTAACGGCAGACTCTGATGGCACGGAGGATGCTCAAGCTTGCTGGAGTACCTAAAGGAAGTTCCTGCGTTAATGATTCTCCATGACCCGTAGACTTTTTAATCTCAAAATTTGACTAAGTCACAGGGGGGAACGTGAGATGCTTAAGGGGAGAAAGTTGTTTTGGTTTCCATAGGAGGTCGTGACCTTACTGGTTTTGTATTGATGCTTGGTTTCTTCTTCACTTGCTCCACTGCTTTTTATCATGAGAGGGGAGATTTGCTGAAAATGAAAGGTCATCAGTTGGCCATTCAGGTGAAGAAATTTGGCCAAGGCACTTTTACGACCCTGAGCTTTGACTGAGTTCAACTGCTAAAATGCGGGATTGGCGGCACTCCACGGGGCCAGTCAGCAGCAAAACAGACACTAACTATTAACTTGGACAGTCAAGAAGACACACACGTTGCTATAAATGTATTGTGAGGTATACAGCCTGCACTCGTACCTGCAGCACTGAAATACTTAATTGAAGTTCTCATTTTGTTAGGAAATGCCATATTTTCCGCTGACCCGGGTTAACACTGCTTTATGCGCTGCTCTCCCCCAAGGGAACCATATGATGCTGTCTTACAACTTACAAAAAAATATTTTAACTCTAGCTGTTAAAAACCTGGGTTTATAG-168-CACGGCTTGGGAGTTTACATCTGAGATGAGCTGTATTCTTTCAGTAAAGAATGTCATAGAAGCTTCTAGGGGAATCCAACTGAGGGATATATGATGTTCTCTCTAGATTATCAGAAGTTTACATGAATCAGTGGAATCATTCCCCCTGCAATGAGCAGTGCTCTTAGCCTAAAGGGAATGCCGGGTCACGTGATAATGTAGCCTGAAAGATGAGGGCATGAGGACAATGGTCTCAGGAGTACTCCCGAAGGGTTCCCTCA02265554 1999-03-16CCTAGGCTCCGCACAGCTCCGTCAGTGATTCACTGGTAGTGTCTCAACATGGGGAGGTTAAAGAGTCAGATTATCAAATTAACAGTAGAATGACCAAAACCATTTTCATACGGCCACCCCGCCTAAAGTCAATCAACGGCTGCTGCAGACCGTTGATAACGGATAGTGATGGATTCAGTTGAGGCTTGTATGTTGTAAGTAAAGGAGATCCCAAGTGATTGATGGCCAAGCCCCAAAGATCCCAGTCCACAGTAATTCGGSUBSTITUTE SHEET (RULE 26)4PCT/US97/16718GAATTGAGGG 9660CAGTGGTTTG 9720AAATCACAAA 9780TCAGTTGAGT 9840GTATATTACC 9900ATGACAGAGA 9960TACATGTGGA 10020GTAGCCATGC 10080CTCAGAGGTG 10140GGGTTTTCTG 10200ACTGATGACA 10260AGACTTGAAG 10320ATTGTGTATG 10380TATCGTGACA 10440ACAATCCGGA 10500TGGAAATCTT 10560CTGACAATGT 10620TACCCGAAAG 10680GATGTTTTCC 10740GGAGCTTACC 10800AAGGAAACAG 10860GCTGAAAATC 10920GATGAGCACG 10980CTCAAAGAAA 11040ACAAGTACCA 11100CAGGACCAAG 11160?WO 98/13501ACACTGATCAATCTCAAGAATAAATGAGATCTGTCCTGTAATAAAGTCCCGTCAGAAGCTGAGTAAGGATTACCCAGCACACTTTGTAATCAATTGTTTCTGTCCCAATCAAACAAGGGCATGACCGTTACTCTTGGCTTACAACGACCTTGAATATGAGATCTCAAGAGCACAACAACCTTGTATGTGTTCCATAGTCCAGGGACTGGCTCCTGGATCAAAGGCTTGATTGTCCAATTAGAAATGTCCTATATGTGGGCTCCGGAGAATGTACTGCCTTTTACGGATTGTGTAAGTGACCAATGATCAAGTGGACCATCTGCTTCGTTAATGGCCCTACTCTTAGGCAAATCACATTTTACTCAAGAGCAGCATGCAGTCCTTGCATATCACAATCAATCTTAATAAGGCAGGCTGTTTAATGATTCTCGGGGGACTCTCCAGAGCATCAAACCCAATGGGCATTCCTCTAGTGTCACATCGAGCCAGCTGACTATGAACATTGACAAAGAGGCTAGCTCAATGGAAGCTTAATTGGAGATCCCTCATTTTCCTCATTGCCATCTTCATTAAGCACCATTCGTGCAAGGGGAACCTTAAGAAGGCTACATGTTTGTCTATTATCGCAAGATAATATTGCTATCCCTGAACGTCAACCATGAATGGCACTGTGTCAGAAACAGCCTCACTAATCATTCCTAGACTAGACTCCTTAAAAGGATATGGACAGGCGGGGCAAGAGATGAGGAAGGCAATTCAGAGGAGTCATGTTCGAGGACGGC-169-ACGAGACAGTATGAGACCATTCCAGTGGCTCCCCCGACCTAGTACCCTATCCTATCTATAACAATCAG@CAACGGGAAGCATATTGGCCACAAAAGGAATGTGTATTCTGCAACAATGGCTCCTAAAAGTCCCGGGATGTTGCCCGCTCCTCGGTGATCCTGCCTGAAGAACTGGGCTAGTCAAGAACATTATTCCATGAATATTATAGTAGTCTATTGCGGGGGTTAACCAGGGATGGTCAGTGCAGCTCTATTTACGG02265554 1999-03-16CAGTGCATTTCAGCTTGTTTGCATAAGAGGTGACGCCCATGGGAGGTATACCTGGCTGCTCATAGCCGTATGCTAGAGTATCACCTCAAGATATTATGATGTCAGAGACTTAAAAGCATCGATACAGCAAAGTCATACCCTATTGGGGGGAGTAACATCAGACCCTCCATCGACCCTTACAACTGCAAGGTGACAGTAAAACCTAGGGCAAGGCATGCTGCTCTCGAGTGGCTATTGACAGGCGAGAGCTCCTTGAGGTCSUBSTITUTE SHEET (FIULE 26). mm .....».....——-—-4-...-.—.-..... ._.,.,PCT/US97/16718ATCACGACTG 11220GCACAGAGGC 11280CTTGAGACCT 11340ATCCCGTTAT 11400GAAGGGTATT 11460TATGAGAGCG 11520ACAAAAAGGG 11580ACTAGAGATT 11640GCAAATGASA 11700GGGCTACTTG 11760ATAGTTGATG 11820GAGAGAGGTT 11880ATTCTGATCT 11940CTCCTCACAA 12000ATGAATTATC 12060TCAATTGCTG 12120CAAGTAATGA 12180TCAGCAAATC 12240TTTGTCCTGA 12300GAAGAGGACG 12360GCTCATGAAA 12420GATACCACAA 12480ATAACCAGAT 12540GGAAGAAAGC 12600CTAAGAAGCC 12660CCTGATGTAC 12720?W0 98/13501TAGAATCTAT GCGAGGCCACGATCAGTCAA CTACGGATGGAGGAAACATC ATCCTTGAGATGAAGCTTGC CTTCGTAAGACAGTGTACTC ATGGGCTTACCTAGGCAAAG GGCCAATGTGCGACTAATTT AGCGCATAGGcccrrerccc AGTGGCGAGGCAGATAAGAQ}GGTTGATACTTTTTAGAAAC ATTGTTTCGATTCACGTCGA AACAGATTGTcccGcAAGcT AGAGCTGAGGCTTTAATTGA CAGAGATGCAAATTTGTTAC ATGGTCCACACTATGATTGA CCTGGTAACATAGGGGATGA CGATATCAATTCACTATCTA CTTGGGCCAGGACCATCAGG GAAATATCAGAAGGAGTGTT TAAGGTGCTTGGCATTGTGG TATTATAGAGCAACTGTGTG CAACATGGTTAAGAGTTAGA AGAGTTCACAGATTCGACAA CATCCAGGCAGGACCTGCCC ACCAATTCGAATATCAAGGC AGAGGCTATGTTGTAGACCA TTACTCATGCCACTTATTCGGCTTTTTTGTCCGTCCCATATAGCCCCAAGTCGGTGATGATGAGCCTGGAGGTTGAGGGATCTATACCACAAAACTTTATATCTCGAGAAAGTGCGTGATCCGCAGAGCTATACAAGGCTATCCCCAACTATAAATTTGAGAAGTTTCATAATGTGCGGCCAATGGGTGAGCGTCAATGCTCCCTATCCATGTACACATGCTTTTCTCTTGTAAACACTTATGGTCTAAGACTTATCTCCAGTCTCTGACTT-170-GTCATGAGACCCTCGGGTTGTTGGTTCTACGATCCTTGCGATAGCTCTTGAGCTAAGGGTGTAGCACTCATCTCCAACGAACCAACAAGGATACCGGATCCGATGATAGAGTACCAACCCACACCCAGAGATCACATTTTAGGACCATATCTGAGTTTCTTCAATTGGGCTGTTGTCATCTAAGCCACCCGTCCTTCACTATATGACCTAGTGAAAGCGAGTGTTCTGGCCGGTAGAGAACAGGATCTTCATCTCCGGCG02265554 1999-03-16ATGTGTCATCCCAACTGGATCACTGATGAGATCTGCTGTTGAACGAAGCCGATCACTCCCAGTGAAATACCAATCTCTCAAATGCTTCTAATCTAACACGTCATCCCAGGATTGATATATCCATAGGAGGAGCTAAGTCCGAATGAAATTGCTCATAGAGATTTGATGTAGTTCCTTTCTAAAGATCTACTGATGCTCAACCTCGACCTGCGAGGATGTAAGATTTGTACATGTGCAGTTGTGGAACATAAGGATCGATCSUBSTITUTE SHEET (RULE 26)PCT/US97/16718TGCGAGTGTG 12780GATATTGACA 12840AGAACAGACA 12900AGAATAGCAA 12960TGGTTGTTGG 13020ATCTCAACTT 13080TCAGGTACAT 13140TTTGTCATAT 13200GGGTTGGGTG 13260GTATTACATC 13320ATACCCAGCT 13380GATAATGCAC 13440CACCTTGTGG 13500ACAGCACTAT 13560TCAGCTCTCA 13620CCAAGATTAT 13680CATTATCATA 13740AGAATGAGCA 13800AAGAAATTCT 13860AACTTGCACA 13920TTGTTGAATG 13980GTACCGGACA 14040TGTCAACCAG 14100CTAACCGACC 14160AATCCAATTA 14220AAACAGATAA 14280?WO 98113501GATTGAGAGTCAAAGATCGGATGATGTTGCGGGGCAATCTCTTGCTACAAACGGCTTGTTAACTAAACAAAATTAGCACCTTGTCAAAGTTCAATTTCATAGACCTTGCCTGGCTCTGCTGGGATTTTGTTATACCCTAGAGGCTAACCGGGACTTCACCCAATTGTGGGCTATAGAGCAAATTGATCCATCTACAGGGACTTACCCCGTTCTGGGGGCAATCTCAAGTCCCAAGTCAGATAACAGTCAAACTAATTGGTTGATCCAGGACAGCAACAACAAAATTGCTCCGCCAATTATAGCTGTTGAGCTTGGGTGAGGTGCTTCTATCTATCCCTCCGCTCTTTAACAGTTAGTAATTGACAAAGATCCTGGGCAAATCAGGGATTTATACAGCAACGCTAATGAATTGGACTTATAAGACGCAGTTGGTGCTGATCCCATGATGTTGTTGGCAAGAATTGGTAAGTCATTCTTCTTCGGCTATCTGGAAACAGATTGGAGACCAAATGAACTCCGGCATTCATTTTCGATCTCAAATAAAAGATATCAGAAATCCATGATATCAACATGGATCGGGTTAATAGTGGGGGAAGTTGGCCGGGAGGCCCGATCCCTACCTACTATAGAGAATAGGATCAAATAAGTTATGTTCATCTCTACCTGAAAAGAGGTCACATCCAGTAGAGGTGAATTGCGGGTGCCTCAGGGCTTCAAAGACAAGCAGGCAACTACTCCGGGAATACTAGACTATTATGACGGGAATGGTATAAACCCTAATC-171-ACGCCCTCGCTGAGCATCAAACACAAGCAACTTTCCGCAGTAATTAGGAGCTATGTTGATTTTCCGCCAATTGTCGAACAAAGTCACGTGCTAGTGTGGGAGCTAGAGGATACTGGTGATTAGGGTCTCACTGAATCTTATTAAGCAGCATATCCATTAAATATCAATCCTGGCAATTAAAAGATGGATTACCAAAGAAGGAGAACTTATACAGAAAGTTTACACCAGAAGGGGTTTGAAAGTTAGTCGGCTGCCCTAGG02265554 1999-03-16TGAGGTAAATGGCTTTCAGAGCACAATCTTAATCGGGTTGATGCCTTGAGCACTTATAAGTTCTAGATCTCAGAATGGGAGGTAGGCAGTGTTTATCCATATTGGCAGCCTAAGCTTATGTTATAGAGAATTTGGTTATGGATAATTGAAGCAACTAAGCTACTCTGAAACGGACCTAAGGCTTAATTCTTCAACAAGGGATCTAGGATCGATAAATAAGTATCTTCGTTACGTGAGTGGATACAGTGCCTGGTTAGGCASUBSTITUTE SHEET (RULE 26)PCT/U S97/ 16718GTCAGTCAGC 14340CCCCCACACG 14400CCCATTTCAG 14460AACTCATCTG 14520CCAGGGGAGG 14580GAGATACTTA 14640GGTCAAAGGG 14700GTAGGTAATA 14760GTAGATTGCT 14820TCAGATATAG 14880ATCTTATCGA 14940CCTTTCAGCG 15000GTGAACCTTG 15060ACAGATCTCA 15120TCATCTGTGA 15180TGCATACAAG 15240AAACTTACAC 15300CTGTGCAAAG 15360ATACTCATCC 15420ATGTTCCACG 15480ACCCGCAAAT 15540TTTATCCAGA 15600AAGAATCTAT 15660GTTTTTAAGG 15720CTGATTAAGG 15780TTATTTGCAA 15840?W0 98/ 13501CA— 17202265554 1999-03-16PCT/U S97/ 16718TATATTAAAG AAAACTTTGA AAATACGAAG TTTCTATTCC CAGCTTTGTC TGGT(2) INFORMATION FOR SEQ ID NO:10:(i)(ii)(xi)MetAspArgIleAsn65AlaIleGlyArgIle145TrpSerSEQUENCE CHARACTERISTICS:LENGTH: 2183 amino acidsamino acidSTRANDEDNESS:TOPOLOGY: linear(A)(B)(C)(D)TYPE:MOLECULE TYPE: proteinSEQUENCE DESCRIPTION: SEQ ID NO:10:Asp Ser Leu Ser Val Asn Gln Ile LeuSerValLys50ValHisGluAsnAsp130LysPheValProPro35HisGluSerAspSer115ThrGluGluIleIle20His5ValAlaThrTyrArg Leu LysValHisLys100LeuAsnLysProLysGlyIle85GluTyrSerValPhe165SerSUBSTITUTE SHEET (RULE 26)Asn70ProSerSerArgIle150LeuGlnAsnSerAsn55ValTyrThrLysLeu135AsnPheThrLysLeu40GlyIleProArgVal120GlyLeuTrpHisIle25GluPheLysAsnLys105SerLeuGlyPheThr10ValAspSerSerCys90IleAspGlyValThr170CYBTyrAlaPTOAsnLys75AsnArgLysSerTyr155ValHisProIleThrGln60LeuGlnGluValGlu140MetLysArgGluLeuLeu45MetArgAspLeuPhe125LeuHisThrArgVal His15Glu30TyrCys GlnIle IleSerTyrPhe95LeuLeu110LysGln CysArg GluSer SerGlu Met175Arg HisLeuAlaAsnAsnPro80AsnLysLeuAspGln160ArgThr15894?W0 98/ 13501ProLeuPhe225ThrArgGlyAlaLeu305PheIlePheAsnLeu385ArgAlaHisGlyValVal210GluGluValA81’!Tyr290AsnSerPheArgVal370MetAspAlaGluCys450Phe195AlaLeuThrArgPro275LeuAspIleSer355ArgLysArgAspGln435Phe180PheIleValAlaTyr260ThrGlnCysGluThr340PheLysGlyHisThr420CysMetThrIleLeuMet:245MetTyrLeuPheGly325AspGlyTyrHisGly405IleValProSUBSTITUTE SHEET (RULE 25)CAGlySerMet230ThrTrpGlnArgThr310ThrMetAla390GlyArgAspLeuSerLys215TyrIleLysIleAsp295GluTyrIleProA811375IleSerAsnAsnSer455- 173Ser200GluCysAspLeuVal280IleIleHisHisArg360GlnPheTrpAlaTrp440Leu185Va].SerThrHi ElGluLeu345LeuProCYEIProGln425LynAspGluGlnValArg250AspMetValAspLeu330ThrGluLysGlyPro410AlaSerSer02265554 l999-03- 16LeuHisIle235TyrGlyLeuGluVal315IleGlyAlaValIle395LeuSerPheAspLeuVal220GluThrPheGluLeu300LeuGluGluValIle380IleThrGlyAlaLeu460Ile205TyrGlyGluPhePIO285ArgAspAlaIleThr365ValIleLeuGluGly445Thr190SerTyrArgLeuPro270LeuGlyGlnLeuPhe350AlaTyrA811ProGly430ValMetPCT/US97/16718Arg AspThrLeuMet240LeuLeu255GlyAla LeuSer LeuAla PheAsn Gly320Asp335TyrSer PheAla GluGlu ThrGly Tyr400Leu His415Leu ThrLys PheTyr Leu?ICA 02265554 1999-03-16W0 98/ 13501 PCT/US97/ 16718-174-Lys Asp Lys Ala Leu Ala Ala Leu Gln Arg Glu Trp Asp Ser Val Tyr465 470 475 480Pro Lys Glu Phe Leu Arg Tyr Asp Pro Pro Lys Gly Thr Gly Ser Arg485 490 495Arg Leu Val Asp Val Phe Leu Asn Asp Ser Ser Phe Asp Pro Tyr Asp500 505 510Val Ile Met Tyr Val Val Ser Gly Ala Tyr Leu His Asp Pro Glu Phe515 520 525Asn Leu Ser Tyr Ser Leu Lys Glu Lys Glu Ile Lys Glu Thr Gly Arg530 535 S40Leu Phe Ala Lys Met Thr Tyr Lys Met Arg Ala Cys Gln Val Ile Ala545 550 555 560Glu Asn Leu Ile Ser Asn Gly Ile Gly Lys Tyr Phe Lys Asp Asn Gly565 570 575Met Ala Lys Asp Glu His Asp Leu Thr Lys Ala Leu His Thr Leu Ala580 585 590Val Ser Gly Val Pro Lys Asp Leu Lys Glu Ser His Arg Gly Gly Pro595 600 605Val Leu Lys Thr Tyr Ser Arg Ser Pro Val His Thr Ser Thr Arg Asn610 615 620Val Arg Ala Ala Lys Gly Phe Ile Gly Phe Pro Gln Val Ile Arg Gln625 630 635 640Asp Gln Asp Thr Asp His Pro Glu Asn Met Glu Ala Tyr Glu Thr Val645 650 655Ser Ala Phe Ile Thr Thr Asp Leu Lys Lys Tyr Cys Leu Asn Trp Arg660 665 670Tyr Glu Thr Ile Ser Leu Phe Ala Gln Arg Leu Asn Glu Ile Tyr Gly675 680 685Leu Pro Ser Phe Phe Gln Trp Leu His Lys Arg Leu Glu Thr Ser Val690 695 700Leu Tyr Val Ser Asp Pro His Cys Pro Pro Asp Leu Asp Ala His Ile705 710 715 720Pro Leu Tyr Lys Val Pro Asn Asp Gln Ile Phe Ile Lys Tyr Pro Met725 730 735SUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/ 13501 PCT/US97/16718-175-Gly Gly Ile Glu Gly Tyr Cys Gln Lys Leu Trp Thr Ile Ser Thr Ile740 745 750Pro Tyr Leu Tyr Leu Ala Ala Tyr Glu Ser Gly Val Arg Ile Ala Ser755 760 765Leu val Gln Gly Asp Asn Gln Thr Ile Ala Val Thr Lys Arg Val Pro770 775 780Ser Thr Trp Pro Tyr Asn Leu Lye Lys Arg Glu Ala Ala Arg Val Thr785 790 795 800Arg Asp Tyr Phe Val Ile Leu Arg Gln Arg Leu His Asp Ile Gly His805 810 815His Leu Lys Ala Asn Glu Thr Ile Val Ser Ser His Phe Phe Val Tyr820 825 830Ser Lye Gly Ile Tyr Tyr Asp Gly Leu Leu Val Ser Gln Ser Leu Lys835 840 845Ser Ile Ala Arg Cys Val Phe Trp Ser Glu Thr Ile Val Asp Glu Thr850 855 860Arg Ala Ala Cys Ser Asn Ile Ala Thr Thr Met Ala Lys Ser Ile Glu865 870 875 880Arg Gly Tyr Asp Arg Tyr Leu Ala Tyr Ser Leu Asn Val Leu Lys Val885 890 895Ile Gln Gln Ile Leu Ile Ser Leu Gly Phe Thr Ile Asn Ser Thr Met900 905 910Thr Arg Asp Val Val Ile Pro Leu Leu Thr Asn Asn Asp Leu Leu Ile915 920 925Arg Met Ala Leu Leu Pro Ala Pro Ile Gly Gly Met Asn Tyr Leu Asn930 935 940Met Ser Arg Leu Phe Val Arg Asn Ile Gly Asp Pro Val Thr Ser Ser945 950 955 960Ile Ala Asp Leu Lys Arg Met Ile Leu Ala Ser Leu Met Pro Glu Glu965 970 975Thr Leu His Gln Val Met Thr Gln Gln Pro Gly Asp Ser Ser Phe Leu980 985 990Asp Trp Ala Ser Asp Pro Tyr Ser Ala Asn Leu Val Cys Val Gln Ser995 1000 1005Ile Thr Arg Leu Leu Lys Asn Ile Thr Ala Arg Phe Val Leu Ile HisSUBSTITUTE SHEET (RULE 26)?ICA 02265554 1999-03-16W0 98ll3501 PCT/US97/16718-176-1010 1015 1020Ser Pro Asn Pro Met Leu Lys Gly Leu Phe His Asp Asp Ser Lys Glu1025 1030 1035 1040Glu Asp Glu Gly Leu Ala Ala Phe Leu Met Asp Arg His Ile Ile Val1045 1050 1055Pro Arg Ala Ala His Glu Ile Leu Asp His Ser Val Thr Gly Ala Arg1060 1065 ' 1070Glu Ser Ile Ala Gly Met Leu Asp Thr Thr Lys Gly Leu Ile Arg Ala1075 1080 1085Ser Met Arg Lys Gly Gly Leu Thr Ser Arg Val Ile Thr Arg Leu Ser1090 1095 1100Asn Tyr Asp Tyr Glu Gln Phe Arg Ala Gly Met Val Leu Leu Thr Gly1105 1110 1115 1120Arg Lys Arg Asn Val Leu Ile Asp Lys Glu Ser Cys Ser Val Gln Leu1125 1130 1135Ala Arg Ala Leu Arg Ser His Met Trp Ala Arg Leu Ala Arg Gly Arg1140 1145 1150Pro Ile Tyr Gly Leu Glu Val Pro Asp Val Leu Glu Ser Met Arg Gly1155 1160 1165His Leu Ile Arg Arg His Glu Thr Cys Val Ile Cys Glu Cys Gly Ser1170 1175 1180Val Asn Tyr Gly Trp Phe Phe Val Pro Ser Gly Cys Gln Leu Asp Asp1185 1190 1195 1200Ile Asp Lys Glu Thr Ser Ser Leu Arg Val Pro Tyr Ile Gly Ser Thr1205 1210 1215Thr Asp Glu Arg Thr Asp Met Lys Leu Ala Phe Val Arg Ala Pro Ser1220 1225 1230Arg Ser Leu Arg Ser Ala Val Arg Ile Ala Thr Val Tyr Ser Trp Ala1235 1240 1245Tyr Gly Asp Asp Asp Ser Ser Trp Asn Glu Ala Trp Leu Leu Ala Arg1250 1255 1260Gln Arg Ala Asn Val Ser Leu Glu Glu Leu Arg Val Ile Thr Pro Ile1265 1270 1275 1280Ser Thr Ser Thr Asn Leu Ala His Arg Leu Arg Asp Arg Ser Thr Gln1285 1290 1295SUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/13501 V PCT /US97ll67l8-177-Val Lys Tyr Ser Gly Thr Ser Leu Val Arg Val Ala Arg Tyr Thr Thr1300 1305 1310Ile Ser Asn Asp Asn Leu Ser Phe Val Ile Ser Asp Lys Lys Val Asp1315 1320 1325Thr Asn Phe Ile Tyr Gln Gln Gly Met Leu Leu Gly Leu Gly Val Leu1330 1335 1340Glu Thr Leu Phe Arg Leu Glu Lys Asp Thr Gly Ser Ser Asn Thr Val1345 1350 1355 1360Leu His Leu His Val Glu Thr Asp Cys Cys Val Ile Pro Met Ile Asp1365 1370 1375His Pro Arg Ile Pro Ser Ser Arg Lys Leu Glu Leu Arg Ala Glu Leu1380 1385 1390Cys Thr Asn Pro Leu Ile Tyr Asp Asn Ala Pro Leu Ile Asp Arg Asp1395 1400 1405Ala Thr Arg Leu Tyr Thr Gln Ser His Arg Arg His Leu Val Glu Phe1410 1415 1420Val Thr Trp Ser Thr Pro Gln Leu Tyr His Ile Leu Ala Lys Ser Thr1425 1430 1435 1440Ala Leu Ser Met Ile Asp Leu Val Thr Lys Phe Glu Lys Asp His Met1445 1450 1455Asn Glu Ile Ser Ala Leu Ile Gly Asp Asp Asp Ile Asn Ser Phe Ile1460 1465 1470Thr Glu Phe Leu Leu Ile Glu Pro Arg Leu Phe Thr Ile Tyr Leu Gly1475 1480 1485Gln Cys Ala Ala Ile Asn Trp Ala Phe Asp Val His Tyr His Arg Pro1490 1495 1500Ser Gly Lys Tyr Gln Met Gly Glu Leu Leu Ser Ser Phe Leu Ser Arg1505 1510 1515 1520Met Ser Lys Gly Val Phe Lys Val Leu Val Asn Ala Leu Ser His Pro1525 1530 1535Lys Ile Tyr Lys Lys Phe Trp His Cys Gly Ile Ile Glu Pro Ile His1540 1545 1550Gly Pro Ser Leu Asp Ala Gln Asn Leu His Thr Thr Val Cys Asn Met1555 1560 1565SUBSTITUTE SHEET (RULE 26)?ICA 02265554 1999-03-16W0 98/13501 PCT/U S971 16718-178-Val Tyr Thr Cys Tyr Met Thr Tyr Leu Asp Leu Leu Leu Asn Glu Glu1570 1575 1580Leu Glu Glu Phe Thr Phe Leu Leu Cys Glu Ser Asp Glu Asp Val Val1585 1590 1595 1600Pro Asp Arg Phe Asp Asn Ile Gln Ala Lys His Leu Cys Val Leu Ala1605 1610 1615Asp Leu Tyr Cys Gln Pro Gly Thr Cys Pro Pro Ile Arg Gly Leu Arg1620 1625 1630Pro Val Glu Lys Cys Ala Val Leu Thr Asp His Ile Lys Ala Glu Ala1635 1640 1645Met Leu Ser Pro Ala Gly Ser Ser Trp Asn Ile Asn Pro Ile Ile Val1650 1655 1660Asp His Tyr Ser Cys Ser Leu Thr Tyr Leu Arg Arg Gly Ser Ile Lys1665 1670 1675 1680Gln Ile Arg Leu Arg Val Asp Pro Gly Phe Ile Phe Asp Ala Leu Ala1685 1690 1695Glu Val Asn Val Ser Gln Pro Lys Ile Gly Ser Asn Asn Ile Ser Asn1700 1705 1710Met Ser Ile Lys Ala Phe Arg Pro Pro His Asp Asp Val Ala Lys Leu1715 1720 1725Leu Lys Asp Ile Asn Thr Ser Lys His Asn Leu Pro Ile Ser Gly Gly1730 1735 1740Asn Leu Ala Asn Tyr Glu Ile His Ala Phe Arg Arg Ile Gly Leu Asn1745 1750 1755 1760Ser Ser Ala Cys Tyr Lys Ala Val Glu Ile Ser Thr Leu Ile Arg Arg1765 1770 1775Cys Leu Glu Pro Gly Glu Asp Gly Leu Phe Leu Gly Glu Gly Ser Gly1780 1785 1790Ser Met Leu Ile Thr Tyr Lys Glu Ile Leu Lys Leu Asn Lys Cys Phe1795 1800 1805Tyr Asn Ser Gly Val Ser Ala Asn Ser Arg Ser Gly Gln Arg Glu Leu1810 1815 1820Ala Pro Tyr Pro Ser Glu Val Gly Leu Val Glu His Arg Met Gly Val1825 1830 1835 1840Gly Asn Ile Val Lys Val Leu Phe Asn Gly Arg Pro Glu Val Thr TrpSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/ 13501 PCT/US97/16718—179—1845 1850 1855Val Gly Ser Val Asp Cys Phe Asn Phe Ile Val Ser Asn Ile Pro Thr1860 1865 1870Ser Ser Val Gly Phe Ile His Ser Asp Ile Glu Thr Leu Pro Asp Lys1875 1880 1885Asp Thr Ile Glu Lys Leu Glu Glu Leu Ala Ala Ile Leu Ser Met Ala1890 1895 1900Leu Leu Leu Gly Lys Ile Gly Ser Ile.Leu Val Ile Lys Leu Met Pro1905 1910 1915 1920Phe Ser Gly Asp Phe Val Gln Gly Phe Ile Ser Tyr Val Gly Ser His1925 1930 1935Tyr Arg Glu Val Asn Leu Val Tyr Pro Arg Tyr Ser Asn Phe Ile Ser1940 1945 1950Thr Glu Ser Tyr Leu Val Met Thr Asp Leu Lys Ala Asn Arg Leu Mac1955 1960 1965Asn Pro Glu Lys Ile Lys Gln Gln Ile Ile Glu Ser Ser Val Arg Thr1970 1975 1980Ser Pro Gly Leu Ile Gly His Ile Leu Ser Ile Lys Gln Leu Ser Cys1985 — 1990 1995 2000Ile Gln Ala Ile Val Gly Asp Ala Val Ser Arg Gly Asp Ile Asn Pro2005 2010 2015Thr Leu Lys Lys Leu Thr Pro Ile Glu Gln Val Leu Ile Asn Cys Gly2020 2025 2030Leu Ala Ile Asn Gly Pro Lys Leu Cys Lys Glu Leu Ile His His Asp2035 2040 2045Val Ala Ser Gly Gln Asp Gly Leu Leu Asn Ser Ile Leu Ile Leu Tyr2050 2055 2060Arg Glu Leu Ala Arg Phe Lys Asp Asn Gln Arg Ser Gln Gln Gly Met2065 2070 2075 2080Phe His Ala Tyr Pro Val Leu Val Ser Ser Arg Gln Arg Glu Leu Ile2085 2090 2095Ser Arg Ile Thr Arg Lys Phe Trp Gly His Ile Leu Leu Tyr Ser Gly2100 2105 2110Asn Arg Lys Leu Ile Asn Lys Phe Ile Gln Asn Leu Lys Ser Gly Tyr2115 2120 2125SUBSTITUTE SHEET (RULE 26) _.. . »..,.,‘.,..-.....,.......-.?CA 02265554 1999-03-16WO 98/13501 PCT/US97/16718-180-Leu Ile Leu Asp Leu His Gln Asn Ile Phe Val Lys Asn Leu Ser Lys2130 2135 2140Ser Glu Lys Gln Ile Ile Met Thr Gly Gly Leu Lys Arg Glu Trp Val2145 2150 2155 2160Phe Lys Val Thr Val Lys Glu Thr Lys Glu Trp Tyr Lys Leu Val Gly2165 2170 2175Tyr Ser Ala Leu Ile Lye Asp2180(2) INFORMATION FOR SEQ ID NO:l1:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 15894 base pairs(B) TYPE: nucleic acid(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: RNA (genomic)(xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:ACCAAACAAA GTTGGGTAAG GATAGTTCAA TCAATGATCA TCTTCTAGTG CACTTAGGAT 60TCAAGATCCT ATTATCAGGG ACAAGAGCAG GATTAGGGAT ATCCGAGATG GCCACACTTT 120TAAGGAGCTT AGCATTGTTC AAAAGAAACA AGGACAAACC ACCCATTACA TCAGGATCCG 180GTGGAGCCAT CAGAGGAATC AAACACATTA TTATAGTACC AATCCCTGGA GATTCCTCAA 240TTACCACTCG ATCCAGACTT CTGGACCGGT TGGTGAGGTT AATTGGAAAC CCGGATGTGA 300GCGGGCCCAA ACTAACAGGG GCACTAATAG GTATATTATC CTTATTTGTG GAGTCTCCAG 360GTCAATTGAT TCAGAGGATC ACCGATGACC CTGACGTTAG CATAAGGCTG TTAGAGGTTG 420TCCAGAGTGA CCAGTCACAA TCTGGCCTTA CCTTCGCATC AAGAGGTACC AACATGGAGG 480ATGAGGCGGA CCAATACTTT TCACATGATG ATCCAATTAG TAGTGATCAA TCCAGGTTCG 540GATGGTTCGG GAACAAGGAA ATCTCAGATA TTGAAGTGCA AGACCCTGAG GGATTCAACA 600TGATTCTGGG TACCATCCTA GCCCAAATTT GGGTCTTGCT CGCAAAGGCG GTTACGGCCC 660CAGACACGGC AGCTGATTCG GAGCTAAGAA GGTGGATAAA GTACACCCAA CAAAGAAGGG 720SUBSTITUTE SHEET (RULE 26)?,......“.M.........e.......: W...» ,.W0 98l1350lTAGTTGGTGAAGGACCTCTCGAAACAAACCGATTAGCCAGGACTGCATGAAAATGGGGGAGTGCAGGATCACTCCATGGGGGCAAGAGATGTATCACTGCAGATCAGTAGGTGAGAATGAGAGAAGCCAGCCCATCTTCCCGCAGGACAGCGGAAGAACAACTAGGTGCGAAAACTTAGGGAGCCAATGGCTCAAGGCCGATATCAGACAGGTCTCAGCACGCGGTCAGGAATCTCCAGGGCGGTTAAGGAGCACCCTCTATTTAGATTGCTTACGCCGACAGGATTGCTTTTTATCCTGATTTGCTGGTAACTGCACCCATACCCTCTGAGGTTTGAACGGTAAGGAGGCGAGGATGCAAGCGGTTGGAGCTACCGAGAGGAGAGCTACAACCGGCACATCGAAGGTCAAGGCTCAGACAGAGGCCGAGAACCAGGTCCCAGAAGAGCAAGCCCATCGGACCCAGGACAAACCATGCCTGACCTGGAGACATCAAGCACGAATCCAAGACAGGAGGAGACAGAGAGAAAATTTCATGGTCGGAAATGATATACTATTAAGTGAGTTATCCATACATGGTAACTCTGGAGCTTTTGGCCGATTCAGCTGGAAAGGCTTGTTTCCCAGACAAGTTGGGGGGCAAGAGAAACCGCCCCTAGACAGCTGACGCCCACGGACACCCGGCCAGAACAACACAGCCGCGGCACGCCATCTCACTGGCCGGAGCGAGCCCTCAGCAATTGAGCGATGACTGGGTTACAGTGCTGACTCTCAATGAATCT-181-GGTTGGATGTCTCTAATCCTGTGACATTGATTGGGATAGACACTTGAGTCTCCTGGAGAAATGCCATGGGCTTACTTTGAAGGTCAGTTCCAGAGATTGCCCCAAGTILTCAGGAAGATAGGGCCCAGCAGTTGACACTGCTGCTTAGGCTCTATAGTGTAACATCCGCCTCAGCCCATCAGTCAAAAACGATCGAGGAAGACCTGCAGGGGGATCAACTGGACGCTGAAATGTTATTACGATCATGGTTCGAAAACAGCG02265554 1999-03-16GGTGAGGAACGGATATCAAGTACATATATCAACTATGTATCTTGATGAACCTCAATTCAGAGTAGGAGTGTCCAGCATATCACATTGGCAAATGCATACTATTTCTACACGAGGGTCAAAAGCAAGTGATAACGGAGTCCGCAAGCCATGCAATGACAGAACCATCCATCACCATCCACTGACTGGAATGCTATGGCAGCAAGAGAAGGCAAGGCGGTGCCTTTGGGAATTTTATGATCAAATCAGGCCTATGTGGATATSUBSTITUTE SHEET (RULE 25)PCT/US97/16718AGGATTGCCG 780AGAACACCCG 840GTAGAGGCAG 900CCTGCTCTTG 960CTTTACCAGC 1020AACAAGTTCA 1080GAACTTGAAA 1140TTTAGATTAG 1200TCTGAACTCG 1260ACTGAGGACA 1320GGTGATCAAA 1380CAGAGTCGAG 1440GCGAGAGCTG 1500AGCCAAGATC 1560GCAGGAATCT 1620AATCTTCTAG 1680ATTGTTATAA 1740CCCACGATTG 1800CATCCGGGCT 1860ATGGTCAGAA 1920AGGCAGTTCG 1980ACCTCGCATC 2040CCCCCCAAGA 2100CAGCGGTGAA 2160TGATGGTGAT 2220TGGCGAACCT 2280?W0 98/ 13501GATACCGAGGGCTTCTGATGAGAGGCAACAGGTAGGGCCATTTGGAACGGCCCTCGGAACGCCGCACTGAAATAATGAAGAAAACAGCCTCTGCTGTTATAGCATATCCAAAGGATCCCAGGCAGAGATTCTCCAAGGAACTAAAGCCGAGCATCACGCACGTTACCTGACAGATGCTGACCAGTCGACCGCCTCCCAAGAAGGGTCGATTCAGAGTCATTGCTGGGGGTTCCTGCCCTTCTGAGCTTGAACAACACCCCGATATGCTATTTGAAACTGCACTTTCCGAAGCACTTCCGGAGATCGCGTCCATCAGGGCCTACAGGAGTGAAGGGGGAGATGGCCAAAATTGAAGGGAGACCCTGGAAGGACGACCCCACCAGGCCGAGCTGACAAATGGTCGGGAAAAAGTGTAATCCGTGACTCTCCTTGAAGATAATCAACTAGTACGTCCACAATGCGCTCCGATAAGATCCTGGTTGTTGAGGACAGGTGTTGGCCATAGTTGTTACTAACTCTCCACACTGACCGGAGAAGGAGGGGCTTGGGAAAGACACCCATTTTTATTGACAAGGTGCACCTGACACCCGAACTATTATGATACACGAGGATAGTTGAGTCAACACCTCTCATGCAGATGTCACTGGCCGAAACGGACCAGTG2TGAGCTCACTCCATTATATGATGATATCAATGAAGTAGAACCTAAATCACAGAGACCTCAACCCACCACTAGGCGACAAGCGATTCCCAGATCCACAGAGACGTACAGCTCACACCTT-182-GGATCTGCTCGAGATCCACGACTCTCAATGAAAAAGGGCAGGTGGTGCAAGCGGGGAATGTCTGGTAOCAGATGAGCTGTAATCAGAAGAATTAAGAAGCAGCATCATGAGAAATCAATCGTTCTCAAGATCCAGAGGACGCCGTCGGGTAAATCCAGCCAAAGGAGCCACTACAGCTCACATTATAAAAACGACTTCGACCTACAGTGAGGAAGGATGATAGGGCCTCCCAAAGCCCGACAGGGCTCAAGGAGAAAGGT02265554 1999-03-16CCATCTCTATAGCTCCTGAGTTCCTCCGCCCAGACGCGAGCCCAATGTGCTCCCCGAGTGCAATCTCCCCTCTCTGATGTTAATCTCCAAAGATCAACAGTCGCCATTCCCCGACTTGAAAACCCGTTGCAGCTGCTGAATTGTTCCTGAGGCTAGAGGAATGATCTTGCACTTACCTGCAACTTAGGAGCAAGTCGGCATGGCAGGCTGATGCTTTATGAATCGGGCGAAAAACTCCTCTGAAAAACTGCCTAACAACASUBSTITUTE SHEET (RULE 26)IPCT/US97/16718GGGGTTCAGG 2340ACTCCAATCC 2400CCCGGACCCC 2460ATTAGCCTCA 2520TCGAAAGTCA 2580TGTGAGCAAT 2640GAGATCCCAG 2700CCAAGATATT 2760GCTAGAATCA 2820GCAAAATATC 2880TGGACTTGGG 2940ACCCATCATA 3000CAGCCGACAA 3060GGAATTTCAG 3120CACCGGCCCT 3180GGATCGGAAG 3240CAAGTTCCAC 3300CAACCCCATG 3360CAAAGTGATT 3420TGGGACATCA 3480GTGCCCCAGG 3540TACATGTTTC 3600GCATTTGGGT 3660AAAGAGGCCA 3720GTGTTCTACA 3780GGGAGTGTCT 3840?. ..4.............-..u.W0 98/13501TCAACGCAAATCCGTGTTGTGAAGAATGCTGGATTGACAACAACATTTATATTATTGCAAGCACCAGTCTGGTTCAAGAATCTGGAGGAGAAGAATTCCGTGTAGACCGTGCCCGGACAAGCCGACGGCACCACCAGCCATGCCCCCGATATTGGAAGGCGACCGAGGTGACTAAACAAACGGTGCCGCGCCCCGGTGCCAATCCAAGACGAGGAAGCCCGGGCCACCAGACCCCAGCCCCGAAGGACCCCTCCTCCTCTCCAAGTGTGCTTATATGAGCGGAATTCAGAGGCGATAGGCGGTCCACATCAATGAAAATCTCACATTAGAGACCTTATGTCAGATGCAAGCATTTACGACAGTGCCCAGCAAAAGCCCCCAGCGCGAACACCCCAATCTGCCAAACCACCCCCTCCCCCTACCCAACCGCACTTAGGGCCCCCCCAACCCCACAGGCAGGGGGGGGGCCCACCCACCCCACTCCCAGACTCGATCCGGCGCCGAACCGCATCTCGAAGGGCAAATGCGGTTAATCACCCGTCTCGGTCAATGCCTGGGAAGAGGGAACTTCAGAAAAGATGGAGCACAGGCATACCCGCTGAATAGTAAGAAGACGTGATCAAATGCCCGAATCCGAAAGACCCAGGCGGCCCATCCTCCTCAACCGCATCCCTTCCTCAACAGGCATCCGAAAGGAACATACCGACAACCAGACACCAACCCCCCAAAAAACACACGACCACGGCCATCACGGGAGCCACCAAGGACATCAACCAAAAGAT— 183 —ATCTGATACCTTTCGGATAACAGTGGCCTTTCATCGACAAGGAGAAAGAAGCCTGGTTTTAAATGAGCAATGGATATCAATCCAGGCAGTTAAATGATGAAACGACCCCCTCCACGGACCCCAGCACAGAGTGGGACCCCCCACCACCCCACAAGAACTCCTCCCTAGACCACACCCAACGAGGGAGCCCCCCGAACAGAAGGCCCCCAGCGGCAACCAACCCGCAGAAACAACCCGAACGTATCCCACACAATCCACCA02265554 1999-03-16GCTCGATACCCGGGTATTACCAACCTGCTGTACAGAGCAAGAGTGAAGTCTGCACTTGGTGACTCTCCATTGAAGACCTTTTTGCAGCCACCAAGGACTACTCACAATGAAAGCGAGAGGACAGCCCTGACGAGGACCAACGGGAAAGAACACAACCGAAAGATCCTCTCAGAACCCAGACCAACCAATCCCCAGCACCCGGGCCGACAGACCAGAACCCGGAAAGGCCACAGCACCCAAGCCTCTCCAACACCCGACGASUBSTITUTE SHEET (RULE 26)PCT/US97/16718CCGCAGAGGT 3900ACCGTTCCTA 3960GTGACCCTTA 4020CTTCCTGAGG 4080TACTCTGCCG 4140GGGATAGGGG 4200GCACAACTCG 4260AATCGATTAC 4320TCAGTTCCTC 4380TTCAAAGTTC 4440CAGCCAGAAG 4500CCAGCCAGCA 4560CACAAGGCCA 4620CCCCCAAGGC 4680ACCCCCAGCA 4740CCGCACAAGC 4800TCCCCGGCAA 4860CCCCGGTCCA 4920CCGCCGGCTC 4980AACCATCGAC 5040CCAGCACCGC 5100AGACCACCCT 5160CAACCCGCGC S220GAGCGATCCC 5280GTCCCCCGGT 5340CACTCAACTC 5400?W0 98/ 13501CCCACCCCTAGGTCTCAAGGACCGGTCAAAAGCTACAAAGATAACTCTCCACAGTTTTGGCAGAGTGTAGGCCCTAGGCGCTGAACTCTCGAGACAATCAATCAATAATGCTCGGGCTCACGGGACCCCAATCAATAAGGAGCGGAGGAAAGTATAGCCTGTCTCGTACACAAGGGTACCGTGTGCAGCCTACACCAAGTTCACAAGGGAACGATCATTAGTAGTCGAGGTACTTGCACAAATCTGGGGACAGATATTGAAAGGAGACACTGAACGTCTCTCCATTGGGGTTATGACTCGTCAATAACTGAACCAATTAGCTTCAAGTAGTTGCCACAGCAAGCCATCGAGACAAGCAGGAGCTGATACCAATTGCTCAGTATCTGCGGATGTTAGAAAATAAAGGCCCGATCCGACGCTACATAGGCTCTTATCTCGAAAAAATGCCTTCCTGTGCTCGACCTAATAGCATCAAGACCCTGAACGGCGTGAATTGACCTATGCAATTGCGGAGTATGAACACGGGAATCCCTGCCATATTCCAATCTCTCTTTCCAGCCATCACGAGGGTAAGATGCACTTGAGACACAAGTGCTCAGATACAATCTGAGAGCAGGAGATGGTCTATGAACATACTATACAGATATCTATCGCTCGGATACGATAACTCACGTCCGAGATTTCAAGAGTGGTTTTGATGAGGTACCCGATGTACACTCGTACAATTGTGCATGACAAGATCGACCATCCAAcssrccrcccTAAGTTGGAGAGGTTTATCG-184-AGAATCAAGAATGGCAGTACAAGATAGGGGCAATCATTAGGAGATTGCAGAATGCAATGAAGATTTGCGGACAGCCGGCAGCGAGCCTGGATATTGGCTGCAACTATCTTGAAATCCTGTCAGGCTTTGAAGTGGAGGTGGTCGACACAGAAGGGGGTGATATACCACTGTCATCGTGTAAGTCCTCTGCTCCGGGTCTTTCAATCCTTTCTAACATACAGTCGGGAGCAATATCATTGGGATGCCAAGGAGCACTAGCA02265554 1999-03-16CTCATCCAATTGTTAACTCTTGGTAGGAATTCATAAAATTAATACAGGAGCCCAGAATATGAGTAGTCCTTTGCACTTCAAAACTACTAATTCAGGGTGTGTGATTTAATCATTATTTGGGCTATGCGCTATTTACTGGGAGTCCTACTTTTGTCCACCGTGCCCAAGTACTTTCATGCCTCCAAGAATGTTGGGAACCGGCAAGTGTTATTGCTGCCGAGGAGGTATCCAGAGGTTGGAAATTGTTGGATAGTCTACATSUBSHTUTESHEET(RULE26)PCTIUS97/16718GTCCATCATG 5460CCAAACACCC 5520AGGAAGTGCA 5580AATGCCCAAT 5640ACTACTGAGA 5700AAGACCGGTT 5760GGCAGGTGCG 5820CCAGTCCATG 5880TCAGGCAATT 5940CCAAGACTAC 6000CGGCCAGAAG 6060CCCCAGTTTA 6120TGGAGGAGAC 6180CATCTTAGAG 6240CATTGTCCTC 6300GCTAGAGGGG 6360TGTTGCAACC 6420AGAGGGGACT 6480CCTCCGGGGG 6540GTTCATTTTA 6600CACAACAGGA 6660TCACTGCCCG 6720AGACGCTGTG 6780CGTAGGGACA 6840GTCATCGGAC 6900CCTGATTGCA 6960? “._.WO 98/13501GTGTGTCTTGAACAAAAAGGACATCAAAATCACAAGTCTCTATCTCCGGCATCATCCACATCCCAAGGGATTTGCTGGCTCATTAGACTTTCTAGATGTAAATCATCGGTAATCTCTGACTTGGTGTATCGGCTGCTGAACAATCAGTTCATTCTCAAACATCTATAGTCTAATCTGAGCAGGTGTTATCGCAACCAGTCAGCCCTTTGTCAGCTTCCAGCCCCTTATCATATCGCTGACAATGGAGACACGAGTGGGCAGAGGGTTGATGAGAACAAGTCCTATGTAAGCTCTTCGTCATTCCCTCTGGATGTCACCACAGTAGGATAGGTTCTGTTTGCATCGGGCAGACTAACTCAAGATGAAGTGGAAGATTAAATAACCCGCCAGGAGCTCATGACTAGCTGTCTATGTCGCTGTACTATGACATAGCAAAAGGTAGAAATCCGGAGTAATGATCCACGGGGAAGCTCGTCAAGCACGGATGATCAATCAAGCAATGCTTCCAACCCATTGAAGGCAAGGGATCCCCTGGTATGTCAGTCGCTCTGATCAAGCAACCCCGAACAATAAACGAGACCGTCATTAACAGTCATGTTTCTCCATCTACACTCGAGCATCAGCCTGAGGACTCCTTAATCCAGAGAATCAAATGCATTGGTCAAAGGGAAACCCTGTTAGACCCAGGGAATCAGAGTTGTCGTTTGGGGGCTCAGCAACTGATTCTATCACTAGGTGTCTGCAGTGATAGAAATGGGCTGTAGGCGTGTAAATAACAGGAT-185-GCTTTAATATAGACCAGGCCTCCTCTACAAACCGCACCCATCGGTAGTTAGATAAATGCCAGAACATCTTGAGCTTGATCCGCAGAGATCGGTCAAGGACACCTCAGAGAGGATAGGGAGATTGGATTATGAACTCAACTCTGCTCAGGGCTTGTATTTAGTATGGGGGAACAACTGAGCTCCGGTGTTCTATGGTGGCTAATTCCCTATGAAATCCCCACAGGCTTTACCCCGACAACAGGGTAAAATCTCCTTCATAC02265554 1999-03-16GTTGCTGCAGTAAAGCCTGACTCTTGAAACGCATCAAGCCATCAAAACTTTTCTACAAAGATGATTGATAGGGTTGCTAGCATAAAAGCCGTGCTGACACTTCACTGACCTACGACTTCAGATCAATACTCTACTGGAGACCCACTACAAGGTCGAGGTTACTTACCTAGATGTACCGAGCATATGACAATTGGGGGAGCCAGGGATCAGACCGACATGCCTCTCATCTCCGAACAGATGCAAGCACTCTGGGGTCTTGTSUBSTITUTE SHEET (RULE 26)PCT/US97/16718GGGGCGTTGT 7020TCTTACGGGA 7080ACAAATGTCC 7140CACCTGAAAT 7200AGGGTGCAAG 7260ATAACCCCCA 7320GACCTTATGT 7380CCATTGCAGG 7440TCAGCACCAA 7500CACTCTTCAA 7560TAGTGAAATT 7620GAGATCTCAC 7680GTGCAGATGT 7740CCAGAACAAC 7800TCAGAGGTCA 7860ACAATGTGTC 7920TGGAAAAGCC 7980TGTTTGAAGT 8040ACTATCTTGA 8100TCAAACTCGC 8160GGAAAGGTGT 8220AATCCTGGGT 8280ACAGAGGTGT 8340ACAAGTTGCG 8400GCGAGAATCC 8460CTGTTGATCT 8520?WO 98/13501GAGTCTGACACGGTTCAGGGGCCAATGAAGGGTTAGTCCCAACATACCTAACCTGGTCAATGTGGTTTATGCCTATAAAGCTGGTGCCGTTGGGATGGTGATAGGGCTGCGTGAAATAGACGCTATCTGTATAAGATAGTCTACACTGTGTAAACAATGTCTCATATTCCCGAGGAAGATAGGTTTTCCAAGGACATCAAAGCCCTTTCTCCCATACTTGTGCTAATCTCTGACATTTGACCGCTATGACAACTGATAGAGTTGAGCTTAATGGACCTATAACCTAGCCTTACCTCTTCACCTGCGGAGGGATCTCCAATTACGTTTACAGGGGTCCCCACACTTCTGTGGGCATGGGAGTAGTGAACCACATCAGAATTCAACCAGATCAGCCATCCTGTCAGAACATCGGAAGTTGGGATATCCAAATCCGTGAACTCATGCTTAAGGGGAGAAAGTTGTTTTGGTTTCCATAGGAGGTCGTGACCTTACTGGTTTTGTATTGATGCTTGGTTTCTTCCAAAATCAAAATACAAATCCAATAGGTGTAATCTGTCCCAATTGGATGGTGAATGTTTTGGCGCCCAAGCCGTCGAATTACATGCTTGCGGATCAGCTGCACATCACATGATAAGAAAAACGTTATACCCTGGAGTATGCTCAAGCACCGCCAAIGTCATCATGTAATCAGGCTCAAAAAGGGACACTAACTATTAACTTGGACAGTCAAGAAGACACACACGTTGCTATAAATGTATTGTGAGGTATACAGCCTGCACTCG-186-TGCTTCGGGACCACAACAATCAACACATTGTAAGGAAGCATGTCAAACTCAACCTACGATCTCATTTTCTAGTGGAATGCCTCAGAATCTAGTCACCCGGGTCACCCAGATAGGGTCCAAAAGTTCACCTGAGTCCCTCATAAAAAACGGAGTCCAAGCTATTTATTTAAGGAATTCGCTCACGGCTTGGGAGTTTACATCTGAGATGAGCTGTATTCTTTCAGTAAAGAATGTCATAGAAGCTTCTAGGGGAATCCAAC02265554 1999-03-16TTCGGGCCATGTGTATTGGCGAGTGGATACGGCGAAGACTAGTTCCAATCACTTCCAGGGTACTTTTATCTTCACATGGGGGTGGACATAGAAGATGGAACATCAGGCATGTGGTTCCCCAGATAGCCCGCGCTTACAGCATTTTCCAACTAGGAGTTATCATAGAAGACGTACTCCAAACCTAGGCTCCGCACAGCTCCGTCAGTGATTCACTGGTAGTGTCTCAACATGGGGAGGTTAAAGAGTCAGATTATCAAATTSUBSTITUTE SHEET (RULE 26)PCT/U S97/ 16718TGATCACACA 8580TGACTATCCC 8640CGAGATTCAA 8700GCCATGCCCC 8760TGGTGATTCT 8820TTGAACATGC 8880CTTTTAGGTT 8940ACCAAAAACT 9000TCACTCACTC 9060CCAATCGCAG 9120ACCCACTAGT 9180GTTATGGACT 9240ATAGTTACCA 9300CTGGAGGACC 9360CAAATGATTA 9420CCGGCCCACT 9480AAAGAGTCAA 9540GTCAGTGATA 9600GAATTGAGGG 9660CAGTGGTTTG 9720AAATCACAAA 9780TCAGTTGAGT 9840GTATATTACC 9900ATGACAGAGA 9960TACATGTGGA 10020GTAGCCATGC 10080?W0 98/13501TGGAGCCTCTCTTTCCTTAAATGAAGGTACTACATCTGACCAGTAACGGCAGACTCTGATGGCACGGAGGATGCTCAAGCTTGCTGGAGTACCTAAAGGAAGTTCCTGCGTTAATGATTCTCCATGACCCGTAGACTTTTTAATCTCAAAATTTGACTAAGTCACAGGGGGGAACGTGAGACACTGATCAATCTCAAGAATAAATGAGATCTGTCCTGTAATAAAGTCCCGTCAGAAGCTGAGTAAGGATTACCCAGCACTTCACTTGCTCCACTGCTTTTTATCATGAGAGGGGAGATTTGCTGAAAATGAAAGGTCATCAGTTGGCCATTCAGGTGAAGAAATTTGGCCAAGGCACTTTTACGACCCTGAGCTTTGACTGAGTTCAACTGCTAAAATGCGGGATTGGCGGCACTCCACGGGGCCAGTCAGCAGCAAAATCCGGAGAATGTACTGCCTTTTACGGATTGTGTAAGTGACCAATGATCAAGTGGACCATCTGCTTCGTTAATGGCCCTACCATACCTGCAGCACTGAAATACTTAACTGAAGTTCTCATTTTGTTAGGAAATGCCATATTTTCCGCTGACCCGGGTTAACACTGCTTTATGCGCTGCTCTCCCCCAAGGGAACCATATGATGCTGTCTTACAACTTACAAAAAAATATTTTAACTCTAGCTGTTAAAAACCTGGGTTTATAGATGGAAGCTTAATTGGAGATCCCTCATTTTCCTCATTGCCATCTTCATTAAGCACCATTCGTGCAAGGGGAACCTTAAGA-187-TGAGGGATATATGATGTTCTCTCTAGATTATCAGAAGTTTACATGAATCAGTGGAATCATTCCCCCTGCAATGAGCAGTGCTCTTAGCCTAAAGGGAATGCCGGGTCACGTGATAATGTAGCCTGAAAGATGAGGGCATGAGGACAATGGTCTCAGGAGTACTCCCGAAGGGTTCCCTCAACGAGACAGTATGAGACCATTCCAGTGGCTCCCCCGACCTAGTACCCTATCCTATCTATAACAATCAGACAACGGGAAGC02265554 1999-03-16AACAGTAGAATGACCAAAACCATTTTCATACGGCCACCCCGCCTAAAGTCAATCAACGGCTGCTGCAGACCGTTGATAACGGATAGTGATGGATTCAGTTGAGGCTTGTATGTTGTAAGTAAAGGAGATCCCAAGTGATTGATGGCCAAGCCCCAAAGATCCCAGTCCACAGTAATTCGGCAGTGCATTTCAGCTTGTTTGCATAAGAGGTGACGCCCATGGGAGGTATACCTGGCTGCTCATAGCCGTATGCTAGAGTAsuasrrrurs SHEET (RULE 25)PCT/US97/16718CTCAGAGGTG 10140GGGTTTTCTG 10200ACTGATGACA 10260AGACTTGAAG 10320ATTGTGTATG 10380TATCGTGACA 10440ACAATCCGGA 10500TGGAAATCTT 10560CTGACAATGT 10620TACCCGAAAG 10680GATGTTTTCC 10740GGAGCTTACC 10800AAGGAAACAG 10860GCTGAAAATC 10920GATGAGCACG 10980CTCAAAGAAA 11040ACAAGTACCA 11100CAGGACCAAG 11160ATCACGACTG 11220GCACAGAGGC 11280CTTGAGACCT 11340ATCCCGTTAT 11400GAAGGGTATT 11460TATGAGAGCG 11520ACAAAAAGGG 11580ACTAGAGATT 11640?W0 98/13501ACTTTGTAATCAATTGTTTCTGTCCCAATCAAACAAGGGCATGACCGTTACTCTTGGCTTACAACGACCTTGAATATGAGATCTCAAGAGCACAACAACCTTGTATGTGTTCCATAGTCCAGGGACTGGCTCCTGGATCAAAGGCTTGATTGTCCAATTAGAAATGTCCTATATGTGGGCTAGAATCTATGATCAGTCAAAGGAAACATCTGAAGCTTGCCAGTGTACTCCTAGGCAAAGCGACTAATTTCCCTTGTCCGTCTTAGGCAAATCACATTTTACTCAAGAGCAGCATGCAGTCCTTGCATATCACAATCAATCTTAATAAGGCAGGCTGTTTAATGATTCTCGGGGGACTCTCCAGAGCATCAAACCCAATGGGCATTCCTCTAGTGTCACATCGAGCCAGCTGACTATGAACATTGACAAAGAGGCTAGCTGCGAGGCCACCTACGGATGGATCCTTGAGACTTCGTAAGA.ATGGGCTTACGGCCAATGTGAGCGCATAGGAGTGGCGAGGCAAGGCTACATGTTTGTCTATTATCGCAAGATAATATTGCTATCCCTGAACGTCAACCATGAATGGCACTGTGTCAGAAACAGCCTCACTAATCATTCCTAGACTAGACTCCTTAAAAGGATATGGACAGGCGGGGCAAGAGATGAGGAAGGCAATTCAGAGGAGTCATGTTCGAGGACGGCCTTATTCGGCTTTTTTGTCCGTCCCATATAGCCCCAAGTCGGTGATGATGAGCCTGGAGGTTGAGGGATCTATACCACAA-188-ATATTGGCCACAAAAGGAATGTGTATTCTGCAACAATGGCTCCTAAAAGTCCCGGGATGTTGCCCGCTCCTCGGTGATCCTGCCTGAAGAACTGGGCTAGTCAAGAACATTATTCCATGAATATTATAGTAGTCTATTGCGGGGGTTAACCAGGGATGGTCAGTGCAGCTCTATTTACGGGTCATGAGACCCTCGGGTTGTTGGTTCTACGATCCTTGCGATAGCTCTTGAGCTAAGGGTGTAGCACTCATCTCCAACGA02265554 1999-03-16TCACCTCAAGATATTATGATGTCAGAGACTTAAAAGCATCGATACAGCAAAGTCATACCCTATTGGGGGGAGTAACATCAGACCCTCCATCGACCCTTACAACTGCAAGGTGACAGTAAAACCTAGGGCAAGGCATGCTGCTCTCGAGTGGCTATTGACAGGCGAGAGCTCCTTGAGGTCATGTGTCATCCCAACTGGATCACTGATGAGATCTGCTGTTGAACGAAGCCGATCACTCCCAGTGAAATACCAATCTCTCASUBSTITUTE SHEET (RULE 26)PCT/US97/16718GCAAATGAGA 11700GGGCTACTTG 11760ATAGTTGATG 11820GAGAGAGGTT 11880ATTCTGATCT 11940CTCCTCACAA 12000ATGAATTATC 12060TCAATTGCTG 12120CAAGTAATGA 12180TCAGCAAATC 12240TTTGTCCTGA 12300GAAGAGGACG 12360GCTCATGAAA 12420GATACCACAA 12480ATAACCAGAT 12540GGAAGAAAGA 12600CTAAGAAGCC 12660CCTGATGTAC 12720TGCGAGTGTG 12780GATATTGACA 12840AGAACAGACA 12900AGAATAGCAA 12960TGGTTGTTGG 13020ATCTCAACTT 13080TCAGGTACAT 13140TTTGTCATAT 13200?.._......».-....«....«..................~._....~ .‘ HW0 98/13501CAGATAAGAATTTTAGAAACTTCACGTCGACCCGCAAGCTCTTTAATTGAAATTTGTTACCTATGATTGATAGGGGATGATCACTATCTAGACCATCAGGAAGGAGTGTTGGCATTGTGGCAACTGTGTGAAGAGTTAGAGATTCGACAAGGACCTGCCCATATCAAGGCTTGTAGACCAGATTGAGAGTCAAAGATCGGATGATGTTGCGGGGCAATCTCTTGCTACAAACGGCTTGTTAACTAAACAAAATTAGCACCGGTTGATACTATTGTTTCGAAACAGATTGTAGAGCTGAGGCAGAGATGCAATGGTCCACACCTGGTAACACGATATCAATCTTGGGCCAGGAAATATCAGTAAGGTGCTTTATTATAGAGCAACATGGTTAGAGTTCACACATCCAGGCAACCAATTCGAAGAGGCTATGTTACTCATGCTGATCCAGGACAGCAACAACAAAATTGCTCCGCCAATTATAGCTGTTGAGCTTGGGTGAGGTGCTTCTATCTATCCCTCCCAAACTTTATATCTCGAGAAAGTGCGTGATCCGCAGAGCTATACAAGGCTATCCCCAACTATAAATTTGAGAAGTTTCATAATGTGCGGCCAATGGGTGAGCGTCAATGCTCCCTATCCATGTACACATGCTTTTCTCTTGTAAACACTTATGGTCTAAGACTTATCTCCAGTCTCTGACTTTTCATTTTCGATCTCAAATAAAAGATATCAGAAATCCATGATATCAACATGGATCGGGTTAATAGTGGGGGAAGTTGGCC-189-ACCAACAAGGATACCGGATCCGATGATAGAGTACCAACCCACACCCAGAGATCACATTTTAGGACCATATCTGAGTTTCTTCAATTGGGCTGTTGTCATCTAAGCCACCCGTCCTTCACTATATGACCTAGTGAAAGCGAGTGTTCTGGCCGGTAGAGAACAGGATCTTCATCTCCGGCGACGCCCTCGCTGAGCATCAAACACAAGCAACTTTCCGCAGTAATTAGGAGCTATGTTGATTTTCCGCCAATTGTCGAACA02265554 1999-03-16AATGCTTCTAATCTAACACGTCATCCCAGGATTGATATATCCATAGGAGGAGCTAAGTCCGAATGAAATTGCTCATAGAGATTTGATGTAGTTCCTTTCTAAAGATCTACTGATGCTCAACCTCGACCTGCGAGGATGTAAGATTTGTACATGTGCAGTTGTGGAACATAAGGATCGATCTGAGGTAAATGGCTTTCAGAGCACAATCTTAATCGGGTTGATGCCTTGAGCACTTATAAATTCTAGATCTCAGAATGGGASUBSTITUTE SHEET (RULE 26)PCT/US97/16718GGGTTGGGTG 13260GTATTACATC 13320ATACCCAGCT 13380GATAATGCAC 13440CACCTTGTGG 13500ACAGCACTAT 13560TCAGCTCTCA 13620CCAAGATTAT 13680CATTATCATA 13740AGAATGAGCA 13800AAGAAATTCT 13860AACTTGCACA 13920TTGTTGAATG 13980GTACCGGACA 14040TGTCAACCAG 14100CTAACCGACC 14160AATCCAATTA 14220AAACAGATAA 14280GTCAGTCAGC 14340CCCCCACACG 14400CCCATTTCAG 14460AACTCATCTG 14520CCAGGGGAGG 14580GAGATACTTA 14640GGTCAAAGGG 14700GTAGGTAATA 14760?W0 98/13501TTGTCAAAGTTCAATTTCATAGACCTTGCCTGGCTCTGCTGGGATTTTGTTATACCCTAGAGGCTAACCGGGACTTCACCCAATTGTGGGCTATAGAGCAAATTGATCCATCTACAGGGACTTACCCCGTTCTGGGGGCAATCTCAAGTCCCAAGTCAGATAACAGTCAAACTAATTGGTTATATTAAAGGCTCTTTAACAGTTAGTAATTGACAAAGATCCTGGGCAAATCAGGGATTTATACAGCAACGCTAATGAATTGGACTTATAAGACGCAGTTGGTGCTGATCCCATGATGTTGTTGGCAAGA' ATTGGTAAGTCATTCTTCTTCGGCTATCTGGAAACAGATTGGAGACCAAATGAACTCCGGAAAACTTTGACAGGGAGGCCCGATCCCTACCTACTATAGAGAATAGGATCAAATAAGTTATGTTCATCTCTACCTGAAAAGAGGTCACATCCAGTAGAGGTGAATTGCGGGTGCCTCAGGGCTTCAAAGACAAGCAGGCAACTACTCCGGGAATACTAGACTATTATGACGGGAATGGTATAAACCCTAATCAAATACGAAG-190-AAGTCACGTGCTAGTGTGGGAGCTAGAGGATACTGGTGATTAGGGTCTCACTGAATCTTATTAAGCAGCATATCCATTAAATATCAATCCTGGCAATTAAAAGATGGATTACCAAAGAAGGAGAACTTATACAAAAAGTTTACACCAGAAGGGGTTTGAAAGTTAGTCGGCTGCCCTAGGTTTCTATTCC02265554 1999-03-16GGTAGGCAGTGTTTATCCATATTGGCAGCCTAAGCTTATGTTATAGAGAATTTGGTTATGGATAATTGAAGCAACTAAGCTACTCTGAAACGGACCTAAGGCTTAATTCTTCAACAAGGGATCTAGGATCGATAAATAAGTATCTTCGTTACGTGAGTGGATACAGTGCCTGGTTAGGCACAGCTTTGTC(2) INFORMATION FOR SEQ ID NO:12:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 2183 amino acids(B) TYPE: amino acid(C) STRANDEDNESS:(D) TOPOLOGY: linear(ii) MOLECULE TYPE: protein(xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:SUBSTITUTE SHEET (RULE 26)PCT/US97/16718GTAGATTGCT 14820TCAGATATAG 14880ATCTTATCGA 14940CCTTTCAGCG 15000GTGAACCTTG 15060ACAGATCTCA 15120TCATCTGTGA 15180TGCATACAAG 15240AAACTTACAC 15300CTGTGCAAAG 15360ATACTCATCC 15420ATGTTCCACG 15480ACCCGCAAAT 15540TTTATCCAGA 15600AAGAATCTAT 15660GTTTTTAAGG 15720CTGATTAAGG 15780TTATTTGCAA 15840TGGT 15894?W0 98/13501MetAspArgIleAsn65AlaIleGlyArgIle145TrpSerProL811Phe225ThrArgAspSerValLys50ValHisGluAsnAsp130LysPheValValVal210GluGluValSerProPro35HisGluSerAspSer115ThrGluGluIlePhe195AlaLeuThrArgLeuIle20HisArgValHisLys100LeuAsnLysProLys180PheIleValAlaTyr260SerValAlaLeuGlyIle85GluTyrSerValPhe165SerThrIleLeuMet245MetSUBSTITUTE SHEET (RULE 26)CAValThrTyrLysA511.70ProSerSerArgIle150LeuGlnGlySerMet230ThrTrpAsnAB!)SerAsn55ValTyrThrLysLeu135AsnPheThrSerLys215TyrIleLys- 191GlnLysLeu40GlyIlePIGArgVal120GlyLeuTrpHisSer200GluCysAspLeuIleIle25GluPheLysAsnLys105SerLeuGlyPheThr185ValSerAspAlaIle265Leu10ValAspSerSerCys90IleAspGlyValThr17 0CysGluGlnValArg250.Asp02265554 l999-03- 16TyrAlaProAsnLys75AsnArgLysSerTyr155ValHisLeuHisIle235TY!‘GlyProIleThrGln60LeuGlnGluValGlu140MetLysArgLeuVal220GluThrPheGluLeuLeu45MetArgAspLeuPhe125LeuHisThrArgIle205TyrGlyGluPheValGlu3 0CysIleSerLeuLeu110GlnArgSerGluArg190SerTyrArgLeuPro270PCT/US97l167l8TyrGlnIleTyrPhe95LysCysGluSerMet175HisArgLeuLeuLeu255AlaLeuAlaA311A811Pro80AsnLysLeuAspGln160ArgThrAspThrMet240GlyLeu?CA 02265554 1999-03-16W0 98/13501 PCT/U S97/ 16718-192-Gly Asn Pro Thr Tyr Gln Ile Val Ala Met Leu Glu Pro Leu Ser Leu275 280 285Ala Tyr Leu Gln Leu Arg Asp Ile Thr Val Glu Leu Arg Gly Ala Phe290 295 300Leu Asn His Cys Phe Thr Glu Ile His Asp Val Leu Asp Gln Asn Gly305 310 315 320Phe Ser Asp Glu Gly Thr Tyr His Glu Leu Thr Glu Ala Leu Asp Tyr325 330 335Ile Phe Ile Thr Asp Asp Ile His Leu Thr Gly Glu Ile Phe Ser Phe340 345 350Phe Arg Ser Phe Gly His Pro Arg Leu Glu Ala Val Thr Ala Ala Glu355 360 365Asn Val Arg Lys Tyr Met Asn Gln Pro Lys Val Ile Val Tyr Glu Thr370 375 380Leu Met Lys Gly His Ala Ile Phe Cys Gly Ile Ile Ile Asn Gly Tyr385 390 395 400Arg Asp Arg His Gly Gly Ser Trp Pro Pro Leu Thr Leu Pro Leu His405 410 415Ala Ala Asp Thr Ile Arg Asn Ala Gln Ala Ser Gly Glu Gly Leu Thr420 425 430His Glu Gln Cys Val Asp Asn Trp Lys Ser Phe Ala Gly Val Lys Phe435 440 445Gly Cys Phe Met Pro Leu Ser Leu Asp Ser Asp Leu Thr Met Tyr Leu450 455 460Lys Asp Lys Ala Leu Ala Ala Leu Gln Arg Glu Trp Asp Ser Val Tyr465 470 475 480Pro Lys Glu Phe Leu Arg Tyr Asp Pro Pro Lys Gly Thr Gly Ser Arg485 490 495Arg Leu Val Asp Val Phe Leu Asn Asp Ser Ser Phe Asp Pro Tyr Asp500 505 510Val Ile Met Tyr Val Val Ser Gly Ala Tyr Leu His Asp Pro Glu Phe515 520 525Asn Leu Ser Tyr Ser Leu Lys Glu Lys Glu Ile Lys Glu Thr Gly Arg530 535 540Leu Phe Ala Lys Met Thr Tyr Lys Met Arg Ala Cys Gln Val Ile AlaSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/ 13501 PCT/U S97/ 16718-193-545 550 555 560Glu Asn Leu Ile Ser Asn Gly Ile Gly Lys Tyr Phe Lys Asp Asn Gly565 570 575Met Ala Lys Asp Glu His Asp Leu Thr Lys Ala Leu His Thr Leu Ala580 585 590Val Ser Gly Val Pro Lys Asp Leu Lys Glu Ser His Arg Gly Gly Pro595 600 605Val Leu Lys Thr Tyr Ser Arg Ser Pro Val His Thr Ser Thr Arg Asn610 615 620Val Arg Ala Ala Lys Gly Phe Ile Gly Phe Pro Gln Val Ile Arg Gln625 630 635 640Asp Gln Asp Thr Asp His Pro Glu Asn Met Glu Ala Tyr Glu Thr Val645 650 655Ser Ala Phe Ile Thr Thr Asp Leu Lys Lys Tyr Cys Leu Asn Trp Arg660 665 670Tyr Glu Thr Ile Ser Leu Phe Ala Gln Arg Leu Asn Glu Ile Tyr Gly675 680 685Leu Pro Ser Phe Phe Gln Trp Leu His Lys Arg Leu Glu Thr Ser Val690 695 700Leu Tyr Val Ser Asp Pro His Cys Pro Pro Asp Leu Asp Ala His Ile705 710 715 720Pro Leu Tyr Lys Val Pro Asn Asp Gln Ile Phe Ile Lys Tyr Pro Met725 730 735Gly Gly Ile Glu Gly Tyr Cys Gln Lys Leu Trp Thr Ile Ser Thr Ile740 745 750Pro Tyr Leu Tyr Leu Ala Ala Tyr Glu Ser Gly Val Arg Ile Ala Ser755 760 765Leu Val Gln Gly Asp Asn Gln Thr Ile Ala Val Thr Lys Arg Val Pro770 775 780Ser Thr Trp Pro Tyr Asn Leu Lys Lys Arg Glu Ala Ala Arg Val Thr785 790 795 800Arg Asp Tyr Phe Val Ile Leu Arg Gln Arg Leu His Asp Ile Gly His805 810 815His Leu Lys Ala Asn Glu Thr Ile Val Ser Ser His Phe Phe Val Tyr820 825 830SUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16W0 98/13501 PCT/US97/16718-194-Ser Lys Gly Ile Tyr Tyr Asp Gly Leu Leu Val Ser Gln Ser Leu Lys835 840 845Ser Ile Ala Arg Cys Val Phe Trp Ser Glu Thr Ile Val Asp Glu Thr850 855 860Arg Ala Ala Cys Ser Asn Ile Ala Thr Thr Met Ala Lys Ser Ile Glu865 870 875 880Arg Gly Tyr Asp Arg Tyr Leu Ala Tyr Ser Leu Asn Val Leu Lys Val885 . 890 895Ile Gln Gln Ile Leu Ile Ser Leu Gly Phe Thr Ile Asn Ser Thr Met900 905 910Thr Arg Asp Val Val Ile Pro Leu Leu Thr Asn Asn Asp Leu Leu Ile915 920 925Arg Met Ala Leu Leu Pro Ala Pro Ile Gly Gly Met Asn Tyr Leu Asn930 935 940Met Ser Arg Leu Phe Val Arg Asn Ile Gly Asp Pro Val Thr Ser Ser945 950 955 960Ile Ala Asp Leu Lys Arg Met Ile Leu Ala Ser Leu Met Pro Glu Glu965 970 975Thr Leu His Gln Val Met Thr Gln Gln Pro Gly Asp Ser Ser Phe Leu980 985 990Asp Trp Ala Ser Asp Pro Tyr Ser Ala Asn Leu Val Cys Val Gln Ser995 1000 1005Ile Thr Arg Leu Leu Lys Asn Ile Thr Ala Arg Phe Val Leu Ile His1010 1015 1020Ser Pro Asn Pro Met Leu Lye Gly Leu Phe His Asp Asp Ser Lys Glu1025 1030 1035 1040Glu Asp Glu Gly Leu Ala Ala Phe Leu Met Asp Arg His Ile Ile Val1045 1050 1055Pro Arg Ala Ala His Glu Ile Leu Asp His Ser Val Thr Gly Ala Arg1060 1065 1070Glu Ser Ile Ala Gly Met Leu Asp Thr Thr Lys Gly Leu Ile Arg Ala1075 1080 1085Ser Met Arg Lys Gly Gly Leu Thr Ser Arg Val Ile Thr Arg Leu Ser1090 1095 1100SUBSTITUTE SHEET (RULE 26)I?CA 02265554 1999-03-16W0 98/13501 PCT/US97/16718-195-Asn Tyr Asp Tyr Glu Gln Phe Arg Ala Gly Met Val Leu Leu Thr Gly1105 1110 1115 1120Arg Lys Arg Asn Val Leu Ile Asp Lys Glu Ser Cys Ser Val Gln Leu1125 1130 1135Ala Arg Ala Leu Arg Ser His Met Trp Ala Arg Leu Ala Arg Gly Arg1140 1145 1150Pro Ile Tyr Gly Leu Glu Val Pro Asp Val Leu Glu Ser Met Arg Gly1155 1160 1165His Leu Ile Arg Arg His Glu Thr Cys Val Ile Cys Glu Cys Gly Ser1170 1175 1180Val Asn Tyr Gly Trp Pha Phe Val Pro Ser Gly Cys Gln Leu Asp Asp1185 1190 1195 1200Ile Asp Lys Glu Thr Ser Ser Leu Arg Val Pro Tyr Ile Gly Ser Thr1205 1210 1215Thr Asp Glu Arg Thr Asp Met Lys Leu Ala Phe Val Arg Ala Pro Ser1220 1225 1230Arg Ser Leu Arg Ser Ala Val Arg Ile Ala Thr Val Tyr Ser Trp Ala1235 1240 1245Tyr Gly Asp Asp Asp Ser Ser Trp Asn Glu Ala Trp Leu Leu Ala Arg1250 1255 1260Gln Arg Ala Asn Val Ser Leu Glu Glu Leu Arg Val Ile Thr Pro Ile1265 1270 1275 1280Ser Thr Ser Thr Asn Leu Ala His Arg Leu Arg Asp Arg Ser Thr Gln1285 1290 1295Val Lys Tyr Ser Gly Thr Ser Leu Val Arg Val Ala Arg Tyr Thr Thr1300 1305 1310Ile Ser Asn Asp Asn Leu Ser Phe Val Ile Ser Asp Lys Lys Val Asp1315 1320 1325Thr Asn Phe Ile Tyr Gln Gln Gly Met Leu Leu Gly Leu Gly Val Leu1330 1335 1340Glu Thr Leu Phe Arg Leu Glu Lys Asp Thr Gly Ser Ser Asn Thr Val1345 1350 1355 1360Leu His Leu His Val Glu Thr Asp Cys Cys Val Ile Pro Met Ile Asp1365 1370 1375His Pro Arg Ile Pro Ser Ser Arg Lys Leu Glu Leu Arg Ala Glu LeuSUBSTITUTE SHEET (RULE 26)?ICA 02265554 1999-03-16W0 98/13501 PCT/US97/16718-196-1380 1385 1390Cys Thr Asn Pro Leu Ile Tyr Asp Asn Ala Pro Leu Ile Asp Arg Asp1395 1400 1405Ala Thr Arg Leu Tyr Thr Gln Ser His Arg Arg His Leu Val Glu Phe1410 1415 1420Val Thr Trp Ser Thr Pro Gln Leu Tyr His Ile Leu Ala Lys Ser Thr1425 1430 1435 1440Ala Leu Ser Met Ile Asp Leu Val Thr Lys Phe Glu Lys Asp His Met1445 1450 1455Asn Glu Ile Ser Ala Leu Ile Gly Asp Asp Asp Ile Asn Ser Phe Ile1460 1465 1470Thr Glu Phe Leu Leu Ile Glu Pro Arg Leu Phe Thr Ile Tyr Leu Gly1475 1480 1485Gln Cys Ala Ala Ile Asn Trp Ala Phe Asp Val His Tyr His Arg Pro1490 1495 1500Ser Gly Lys Tyr Gln Met Gly Glu Leu Leu Ser Ser Phe Leu Ser Arg1505 1510 1515 1520Met Ser Lys Gly Val Phe Lys Val Leu Val Asn Ala Leu Ser His Pro1525 1530 1535Lys Ile Tyr Lys Lys Phe Trp His Cys Gly Ile Ile Glu Pro Ile His1540 1545 1550Gly Pro Ser Leu Asp Ala Gln Asn Leu His Thr Thr Val Cys Asn Met1555 1560 1565Val Tyr Thr Cys Tyr Met Thr Tyr Leu Asp Leu Leu Leu Asn Glu Glu1570 1575 1580Leu Glu Glu Phe Thr Phe Leu Leu Cys Glu Ser Asp Glu Asp Val Val1585 1590 1595 1600Pro Asp Arg Phe Asp Asn Ile Gln Ala Lys His Leu Cys Val Leu Ala1605 1610 1615Asp Leu Tyr Cys Gln Pro Gly Thr Cys Pro Pro Ile Arg Gly Leu Arg1620 1625 1630Pro Val Glu Lys Cys Ala Val Leu Thr Asp His Ile Lys Ala Glu Ala1635 1640 1645Met Leu Ser Pro Ala Gly Ser Ser Trp Asn Ile Asn Pro Ile Ile Val1650 1655 1660SUBSTITUTE SHEET (RULE 26)?CAW098/13501- 197 -Asp His Tyr Ser Cys Ser Leu1665 1670 1675Gln Ile Arg Leu Arg Val Asp Pro Gly Phe Ile1685 1690Glu Val Asn Val Ser Gln Pro1700 1705Met Ser Ile Lys Ala Phe Arg Pro Pro His Asp1715 1720Leu Lys Asp Ile1730 1735Asn Leu Ala Asn Tyr Glu Ile His Ala Phe1745 1750Ser Ser Ala Cys Tyr Lys1765Cys Leu Glu Pro Gly Glu Asp Gly Leu Phe1780 1785Ser Met Leu Ile Thr Tyr1795Lys Glu Ile1800Tyr Asn Ser Gly Val Ser1810Ala Asn Ser Arg1815Ala Pro Tyr Pro Ser Glu Val Gly Leu1825 1830ValGly Asn Ile Val Lys Val Leu Phe Asn1845GlyVal Gly Ser Val Asp Cys Phe Asn Phe Ile1860 1865Ser Ser Val Gly Phe Ile His Ser Asp Ile1875 1880Asp Thr Ile Glu Lys Leu Glu Glu Leu Ala1890 1895Leu Leu Leu Gly Lys Ile Gly Ser Ile Leu1905 1910Phe Ser Gly Asp Phe Val Gln Gly Phe Ile1925Asn Thr Ser Lys His Asn Leu1850193002265554 1999-03-16PCTIUS97/ 16718Thr Ty: Leu Arg Arg Gly Ser Ile Lys1680Phe Asp Ala Leu Ala1695Lye Ile Gly Ser Asn Asn Ile Ser Asn1710Asp Val Ala Lys Leu1725Pro Ile Ser Gly Gly1740Arg Arg Ile Gly Leu Asn1755 1760Ala Val Glu Ile Ser Thr Leu Ile Arg Arg17701775Leu Gly Glu Gly Ser Gly1790Leu Lys Leu Asn Lys Cys Phe1805Ser Gly Gln Arg Glu Leu1820Glu His Arg Met Gly Val1835 1840Arg Pro Glu Val Thr Trp1855Val Ser Asn Ile Pro Thr1870Glu Thr Leu Pro Asp Lys1885Ala Ile Leu Ser Met Ala1900Val Ile Lys Leu Met Pro1915 1920Ser Tyr Val Gly Ser His1935SUBSTITUTE SHEET (RULE 25)?ICA 02265554 1999-03-16W0 98/13501 PCT/US97/16718-198-Tyr Arg Glu Val Asn Leu Val Tyr Pro Arg Tyr Ser Asn Phe Ile Ser1940 1945 1950Thr Glu Ser Tyr Leu Val Met Thr Asp Leu Lys Ala Asn Arg Leu Met1955 1960 1965Asn Pro Glu Lys Ile Lys Gln Gln Ile Ile Glu Ser Ser Val Arg Thr1970 1975 1980Ser Pro Gly Leu Ile Gly His Ile Leu Ser Ile Lys Gln Leu Ser Cys1985 1990 1995 2000Ile Gln Ala Ile Val Gly Asp Ala Val Ser Arg Gly Asp Ile Asn Pro2005 2010 2015Thr Leu Lys Lys Leu Th: Pro Ile Glu Gln Val Leu Ile Asn Cys Gly2020 2025 2030Leu Ala Ile Asn Gly Pro Lys Leu Cys Lys Glu Leu Ile His His Asp2035 2040 2045Val Ala Ser Gly Gln Asp Gly Leu Leu Asn Ser Ile Leu Ile Leu Tyr2050 2055 2060Arg Glu Leu Ala Arg Phe Lys Asp Asn Gln Arg Ser Gln Gln Gly Met2065 2070 2075 2080Phe His Ala Tyr Pro Val Leu Val Ser Ser Arg Gln Arg Glu Leu Ile2085 2090 2095Ser Arg Ile Thr Arg Lys Phe Trp Gly His Ile Leu Leu Tyr Ser Gly2100 2105 2110Asn Lys Lys Leu Ile Asn Lys Phe Ile Gln Asn Leu Lys Ser Gly Tyr2115 2120 2125Leu Ile Leu Asp Leu His Gln Asn Ile Phe Val Lys Asn Leu Ser Lys2130 2135 2140Ser Glu Lys Gln Ile Ile Met Thr Gly Gly Leu Lys Arg Glu Trp Val2145 2150 2155 2160Phe Lys Val Thr Val Lys Glu Thr Lys Glu Trp Tyr Lys Leu Val Gly2165 2170 2175Tyr Ser Ala Leu Ile Lys Asp2180(2) INFORMATION FOR SEQ ID NO:13:(i) SEQUENCE CHARACTERISTICS:(A) LENGTH: 15894 base pairsSUBSTITUTE SHEET (RULE 26)?CA 02265554 1999-03-16WO 98/13501-199-(B) TYPE: nucleic acid(C) STRANDEDNESS: single(D) TOPOLOGY: linear(ii) MOLECULE TYPE: RNA (genomic)(xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:ACCAAACAAA GTTGGGTAAG GATAGTTCAA TCAATGKPCA TTTTCTAGTGTCAAGATCCTTAAGGAGCTTGTGGAGCCATTTACCACTCGGCGGGCCCAAGTCAATTGATTCCAGAGTGAATGAGGCGGAGATGGTTCGATGATTCTGGGCAGACACGGCTAGTTGGTGAAGGACCTCTCGAAACAAACCGATTAGCCAGGACTGCATGAAAATGGGGGAGTGCAGGATCACTCCATGGGGGCAAGAGATATTATCAGGGAGCATTGTTCCAGAGGAATCATCCAGACTTACTAACAGGGICAGAGGATCCCAGTCACAACCAATACTTTGAACAAGGAATACCATCCTAAGCTGATTCGATTTAGATTGCTTACGCCGACAGGATTGCTTTTTATCCTGATTTGCTGGTAACTGCACCCATACCCTCTGAGGTTTGAACGGTAAGGAGGACAAGAGCAGAAAAGAAACAAAACACATTACTGGACCGGTGCACTAATAGACCGATGACCTCTGGCCTTATCACATGAIGATCTCAGATAGCTCAAATTTGAGCTAAGAAGAGAGAAAATTTCATGGTCGGAAATGATATACTATTAAGTGAGTTATCCATACATGGTAACTCTGGAGCTTTTGGCCGATTCAGCTGGAAGATTAAGGATAGGACAAACCTTATAGTACCTGGTCAGGTTGTATATTATCCTGACGTTAGCCTTCGCATCATCCAATTAGTTGAAGTGCAGGGTCTTGCTGGTGGATAAAGGTTGGATGTCTCTAATCCTGTGACATTGATTGGGATAGACACTTGAGTCTCCTGGAGAAATGCCATGGGCTTACTTTGAAGGTCAGTTCATCCGAGATGACCCATTACAAATCCCTGGAAATTGGAAACCTTATTTGTGCATAAGGCTGAAGAGGTACCTAGTGATCAAAGACCCTGAGCGCAAAGGCGGTACACCCAAGGTGAGGAACGGATATCAAGTACATATATCAACTATGTATCTTGATGAACCTCAATTCAGAGTAGGAGTGTCCAGCATATCACATTGGCASUBSTITUTE SHEET (RULE 26)PCT/US97/16718CACTTAGGAT 60GCCACACTTT 120TCAGGATCCG 130GATTCCTCAA 240CCGGATGTGA 300GAGTCTCCAG 360TTAGAGGTTG 420AACATGGAGG 480TCCAGGTTCG 540GGATTCAACA 600GTTACGGCCC 660CAAAGAAGGG 720AGGATTGCCG 780AGAACACCCG 840GTAGAGGCAG 900CCTGCTCTTG 960CTTTACCAGC 1020AACAAGTTCA 1080GAACTTGAAA 1140TTTAGATTAG 1200TCTGAACTCG 1260?W0 98/13501GTATCACTGCAGATCAGTAGGTGAGAATGAGAGAAGCCAGCCCATCTTCCCGCAGGACAGCGGAAGAACAACTAGGTGCGAAAACTTAGGGAGCCGATGGCTCAAGGCCGATATCAGACAGGTCTCAGCACGCGGTCAGGAATCTCCAGGGCGGTTAAGGAGCACCCTCTGATACCGAGGGCTTCTGATGAGAGGCAACAGGTAGGGCCATTTGGAACGGCCCTCGGAACGCCGCACTGAAATAATGAAGAAAACAGCCTCGAGGATGCAAGCGGTTGGAGCTACCGAGAGGAGAGCTACAACCGGCACATCGAAGGTCAAGGCTCAGACAGAGGCCGAGAACCAGGTCCCAGAAGAGCAAGCCCATCGGACCCAGGACAAACCATGCCTGACCTGGAGACATCAAGCACGAATCCAAGACAGGAGGKGAGATATGCTATTTGAAACTGCACTTTCCGAAGCACTTCCGGAGATCGCGTCCATCAGGGCCTACAGGAGTGAAGGGGGAGATGGCCAAAATCAAGGCTTGTTTCCCAGACAAGTTGGGGGGCAAGAGAAACCGCCCCTAGACAGCTGACGCCCACGGACACCCGGCCAGAACAACACAGCCGCGGCACGCCATCTCACTGGCCGGAGCGAGCCCTCAGCAATTGAGCGATGACTGGGTTACAGTGCTGACTCTCAATGAATCTCACTGACCGGAGAAGGAGGGGCTTGGGAAAGACACCCATTTTTATTGACAAGGTGCACCTGACACCCGAACTATTATGATACACGAGGAT-200-CAGAGATTGCCCCAAGTATCAGGAAGATAGGGCCCAGCAGTTGACACTGCTGCTTAGGCTCTATAGTGTAACATCCGCCTCAGCCCATCAGTCAAAAACGATCGAGGAAGACCTGCAGGGGGATCAACTGGACGCTGAAATGTTATTATGATCATGGTTCGAAAACAGCGGGATCTGCTCGAGATCCACGACTCTCAATGAAAAAGGGCAGGTGGTGCAAGCGGGGAATGTCTGGTACCAGATGAGCTGTAATCAGAAGA02265554 1999-03-16AATGCATACTATTTCTACACGAGGGTCAAAAGCAAGTGATATCGGAGTCCGCAAGCCATGCAATGACAGAACCCTCCATCACCATCCACTGACTGGAATGCTATGGCAGCAAGAGAAGGCAAGGCGGTGCCTTTGGGAATTTTATGATCAAATCAGGCCTATGTGGATATCCATCTCTATAGCTCCTGAGTTCCTCCGCCCAGACGCGAGCCCAATGTGCTCCCCGAGTGCAATCTCCCCTCTCTGATGTTAATCTCCAASUBSTITUTE SHEET (RULE 26)PCT/US97/16718ACTGAGGACA 1320GGTGATCAAA 1380CAGAGTCGAG 1440GCGAGAGCTG 1500AGCCAAGATC 1560GCAGGAATCT 1620AATCTTCTAG 1680ATTGTTATAA 1740CCCACGATTG 1800CATCCGGGCT 1860ATGGTCAGAA 1920AGGCAGTTCG 1980ACCTCGCATC 2040CCCCCCAAGA 2100CAGCGGTGAA 2160TGATGGTGAT 2220TGGCGAACCT 2280GGGGTTCAGG 2340ACTCCAATCC 2400TCCGGACCCC 2460ATTAGCCTCA 2520TCGAAAGTCA 2580TGTGAGCAAT 2640GAGATCCCAG 2700CCAAGATATT 2760GCTAGAATCA 2820?CA 02265554 l999-03- 16DEMANDES OU BREVETS VOLUMINEUXLA. PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVETCOMPREND PLUS D'UN TOME. ' _.CECI EST LE TOME’ DealNOTE: Pour les tomes additionels, veuillez contacter le Bureau canadien desbrevetsJUMBO APELICATIONSIPATENTSTHIS SECTION OF THE APPLICATIONIPATENT CONTAINS MORETHAN ONE VOLUME 'I-THIS IS VOLUME [_ OF’ 2NOTE: ‘F0-r additional volumes-please contabtlthé Canadian Patent Office

Claims (46)

What is claimed is:

1. An isolated, recombinantly-generated, attenuated, nonsegmented, negative-sense, single stranded RNA virus of the Order Mononegavirales having at least one attenuating mutation in the 3' genomic promoter region and having at: least one attenuating mutation in the RNA polymerase gene.

2. The virus of Claim 1 wherein the virus is from the Family Paramyxoviridae.

3. The virus of Claim 2 wherein the virus is from the Subfamily Paramyxovirinae.

4. The virus of Claim 3 wherein the virus is from the Genus Morbillivirus.

5. The virus of Claim 4 wherein the virus is measles virus.

6. The measles virus of Claim 5 wherein:
(a) the at least one attenuating mutation in the 3' genomic promoter region is selected from the group consisting of nucleotide 26 (A ~ T), nucleotide 42 (A
~ T or A ~ C) and nucleotide 96 (G ~
A), where these nucleotides are presented in positive strand, antigenomic, message sense; and (b) the at least one attenuating mutation in the RNA polymerase gene is selected from the group consisting of nucleotide changes which produce changes in an amino acid selected from the group consisting of residues 331 (isoleucine ~ threonine), 1409 (alanine ~
threonine), 1624 (threonine ~ alanine), 1649 (arginine ~ methionine), 1717 (aspartic acid ~ alanine), 1936 (histidine ~ tyrosine), 2074 (glutamine ~ arginine) and 2114 (arginine ~ lysine).

7. The virus of Claim 3 wherein the virus is from the Genus Paranyxovirus.

8. The virus of Claim 7 wherein the virus is human parainfluenzae virus type 3 (PIV-3).

9. The PIV-3 of Claim 8 wherein:
(a) the at least one attenuating mutation in the 3' genomic promoter region is selected from the group consisting of nucleotide 23 (T ~ C), nucleotide 24 (C
~ T), nucleotide 28 (G ~ T) and nucleotide 45 (T ~ A), where these nucleotides are presented in positive strand, antigenomic, message sense; and (b) the at least one attenuating mutation in the RNA polymerase gene is selected from the group consisting of nucleotide changes which produce changes in an amino acid selected from the group consisting of residues 942 (tyrosine ~
histidine), 992 (leucine ~
phenylalanine), and 1558 (threonine ~
isoleucine).

10. The virus of Claim 3 wherein the virus is from the Genus Rubulavirus.

11. The virus of Claim 2 wherein the virus is from the Subfamily Pneumovirinae.

12. The virus of Claim 11 wherein the virus is from the Genus Pneumovirus.

13. The virus of Claim 12 wherein the virus is human respiratory syncytial virus (RSV) subgroup B.

14. The virus of Claim 13 wherein:
(a) the at least one attenuating mutation in the 3' genomic promoter region is selected from the group consisting of nucleotide 4 (C ~ G) and the insertion of an additional A in the stretch of A's at nucleotides 6-11, where these nucleotides are presented in positive strand, antigenomic, message sense; and (b) the at least one attenuating mutation in the RNA polymerase gene is selected from the group consisting of nucleotide changes which produce changes in an amino acid selected from the group consisting of residues 353 (arginine ~
lysine), 451 (lysine ~ arginine), 1229 (aspartic acid ~ asparagine), 2029 (threonine ~ isoleucine) and 2050 (asparagine ~ aspartic acid).

15. The virus of Claim 1 wherein the virus is from the Family Rhabdoviridae.

16. The virus of Claim 1 wherein the virus is from the Family Filoviridae.

17. A vaccine comprising an isolated, recombinantly-generated, attenuated, nonsegmented, negative-sense, single stranded RNA virus of the Order Mononegavirales according to Claim 1 and a physiologically acceptable carrier.

18. The vaccine of Claim 17 comprising a measles virus according to Claim 5 and a physiologically acceptable carrier.

19. The vaccine of Claim 18 comprising a measles virus according to Claim 6 and a physiologically acceptable carrier.

20. The vaccine of Claim 17 comprising a PIV-3 according to Claim 8 and a physiologically acceptable carrier.

21. The vaccine of Claim 20 comprising a PIV-3 according to Claim 9 and a physiologically acceptable carrier.

22. The vaccine of Claim 17 comprising an RSV subgroup B according to Claim 13 and a physiologically acceptable carrier.

23. The vaccine of Claim 22 comprising an RSV subgroup B according to Claim 14 and a physiologically acceptable carrier.

24. A method for immunizing an individual to induce protection against a nonsegmented, negativesense, single stranded RNA virus of the Order Mononegavirales which comprises administering to the individual the vaccine of Claim 17.

25. The method of Claim 24 wherein the vaccine is the vaccine of Claim 18.

26. The method of Claim 25 wherein the vaccine is the vaccine of Claim 19.

27. The method of Claim 24 wherein the vaccine is the vaccine of Claim 20.

28. The method of Claim 27 wherein the vaccine is the vaccine of Claim 21.

29. The method of Claim 24 wherein the vaccine is the vaccine of Claim 22.

30. The method of Claim 29 wherein the vaccine is the vaccine of Claim 23.

31. An isolated nucleic acid molecule comprising a measles virus sequence in positive strand, antigenomic message sense selected from the group consisting of 1977 wild-type strain (SEQ ID NO:3), 1983 wild-type strain (SEQ ID NO:5) where the nucleotide 2499 is G or C, Montefiore wild-type strain (SEQ ID
NO:7), Rubeovax TM vaccine strain (SEQ ID NO:9), where the nucleotide 2143 is T or C, Moraten vaccine strain (SEQ ID NO:11), Schwarz vaccine strain (SEQ ID NO:11), where the nucleotide 4917 is C and the nucleotide 4924 is C, and Zagreb vaccine strain (SEQ ID NO:13), and the complementary genomic sequences thereof.

32. An isolated nucleic acid molecule comprising a PIV-3 sequence in positive strand, antigenomic message sense selected from the group consisting of cp45 vaccine strain grown in fetal rhesus lung cells (SEQ ID NO:19) and cp45 vaccine strain grown in Vero cells (SEQ ID NO:21), and the complementary genomic sequences thereof.

33. A composition which comprises a transcription vector comprising an isolated nucleic acid molecule encoding a genome or antigenome of a nonsegmented, negative-sense, single stranded RNA virus of the Order Mononegavirales having at least one attenuating mutation in the 3' genomic promoter region and having at least one attenuating mutation in the RNA
polymerase gene, together with at least one expression vector which comprises at least one isolated nucleic acid molecule encoding the trans-acting proteins necessary for encapsidation, transcription and replication, whereby upon expression an infectious attenuated virus is produced.

34. The composition of Claim 33 wherein the transcription vector comprises an isolated nucleic acid molecule which encodes a measles virus according to Claim 5 and the at least one expression vector comprises at least one isolated nucleic acid molecule encoding the trans-acting proteins N, P and L.

35. The composition of Claim 34 wherein the transcription vector comprises an isolated nucleic acid molecule which encodes a measles virus according to Claim 6.

36. The composition of Claim 33 wherein the transcription vector comprises an isolated nucleic acid molecule which encodes a PIV-3 according to Claim 8 and the at least one expression vector comprises at least one isolated nucleic acid molecule encoding the transacting proteins NP, P and L.

37. The composition of Claim 36 wherein the transcription vector comprises an isolated nucleic acid molecule which encodes a PIV-3 according to Claim 9.

38. The composition of Claim 33 wherein the transcription vector comprises an isolated nucleic acid molecule which encodes an RSV subgroup B according to Claim 13 and the at least one expression vector comprises at least one isolated nucleic acid molecule encoding the trans-acting proteins N, P, L and M2.

39. The composition of Claim 38 wherein the transcription vector comprises an isolated nucleic acid molecule which encodes an RSV subgroup B according to Claim 14.

40. A method for producing infectious attenuated nonsegmented negative-sense single stranded RNA virus of the Order Mononegavirales which comprises transforming or transfecting host cells with the at least two vectors of Claim 33 and culturing the host cells under conditions which permit the co-expression of these vectors so as to produce the infectious attenuated virus.

41. The method of Claim 40 wherein the virus is the measles virus of Claim 5.

42. The method of Claim 41 wherein the virus is the measles virus of Claim 6.

43. The method of Claim 40 wherein the virus is the PIV-3 of Claim 8.

44. The method of Claim 43 wherein the virus is the PIV-3 of Claim 9.

45. The method of Claim 40 wherein the virus is the RSV subgroup B of Claim 13.

46. The method of Claim 45 wherein the virus is the RSV subgroup B of Claim 14.