CA2406999A1 - Gene and sequence variation associated with sensing carbohydrate compounds and other sweeteners - Google Patents

Abstract

The present invention relates to the discovery of a gene and its sequence variation associated with preference for carbohydrates, other sweet compound s, or ethanol. The present invention also relates to the study of metabolic pathways to identify other genes, receptors, and relationships that contribu te to differences in sensing of carbohydrates or ethanol. The present invention also relates to germline or somatic sequence variations and its use in the diagnosis and prognosis of predisposition to diabetes, other obesity related disorders, or ethanol consumption. The present invention also provided probe s or primers specific for the detection and analysis of such sequence variatio n. The present invention also relates to method for screening drugs for inhibition or restoration of gene function as antidiabetic, antiobesity, or antialcohol consumption therapies. The present invention relates to other antidiabetic, antiobesity disorder, or antialcohol consumption therapies, su ch as gene therapy, protein replacement therapy, etc. Finally, the present invention relates to a method for identifying sweeteners or alcohols utilizi ng the gene and its variations.

Description

GENE AND SEQUENCE VARIATION ASSOCIATED WITH SENSING
CARBOHYDRATE COMPOUNDS AND OTHER SWEETENERS
FIELD OF THE INVENTION
The present invention relates generally to the field of mouse and human genetics and sensing of extracellular carbohydrates. Specifically, the present invention relates to the discovery of a gene and its sequence variation associated with a differential preference for sweet compounds in laboratory strains of mice.
BACKGROUND OF THE INVENTION
The ability to sense extra-cellular carbohydrates, transduce this sensory information, and relay it to the brain, is carried out by membrane bound receptors in taste papillae. Many approaches to identify the sweet receptor or receptors have been tried, but the problem has proved, until recently, to be difficult.
Mammals vary in their ad libitum consumption of sweeteners. To investigate the genetic contribution to this complex behavior, behavioral, electrophysiological, and genetic studies were conducted using two strains of mice that differ markedly in their preference for sucrose and saccharin (Bachmanov et al., Behavior Genetics, 1996;26:563-573).
Recently published data indicates that the ability to sense carbohydrates is linked to obesity. These studies demonstrated that sensation of simple carbohydrates is suppressible by the adipose hormone, leptin.
These studies demonstrated that a locus on the telomere of mouse chromosome 4 accounts for ~40% of the genetic variability in sucrose and saccharin intake, and that the effect of this locus is to enhance or retard the gustatory neural response to sucrose.

SUMMARY OF THE INVENTION
The present invention provides a gene and its sequence variation associated with a preference for carbohydrate compounds, other sweeteners, or alcohol.
The present invention provides a gene and its sequence variation associated a differential response by the pancreas and/or muscle in response to dietary carbohydrates.
The present invention also relates to sequence variation and its use in the diagnosis and prognosis of predisposition to diabetes, other obesity-related disorders, or alcohol consumption.
The present invention also relates to the study of taste to identify molecules responsible for signal transduction, other receptors and genes and relationships that contribute to taste preference.
The present invention also relates to the study of diabetes to identify molecules responsible for sensing extra-cellular carbohydrate, other receptors and genes and relationships that contribute to a diabetic state.
The present invention also relates to a sequence variation and its use in the identification of specific alleles altered in their specificity for carbohydrate compounds.
The present invention also relates to a recombinant construct comprising SACl (also referred to as Sac) polynucleotide suitable for expression in a transformed host cell.
The present invention also provides primers and probes specific for the detection and analysis of the SACl locus.
The present invention also relates to kits for detecting a polynucleotide comprising a portion of the SAC 1 locus.
The present invention also relates to transgenic animals, which carry an altered SAC1 allele, such as a knockout mouse.
The present invention also relates to methods for screening drugs for inhibition or restoration of SACl function as a taste receptor.

The present invention also relates to identification of sweeteners or alcohols using the SACl gene and its sequence variations.
The present invention also relates to methods for screening drugs for inhibition or restoration of SAC1 function in homeostatic regulation of glucose levels.
The present invention also relates to methods for screening drugs for modification of SAC1 function in the consumption of alcohol.
Finally, the present invention provides therapies directed to diabetic or obesity disorders. Therapies of diabetes and obesity include gene therapy, protein replacement, protein mimetics, and inhibitors.
BRIEF DESCRIPTION OF THE FIGURES
Fig. 1A shows genetic mapping of the SAC1 locus, using 632 F2 mice from a cross between the B6 (high preference) and 129 (low preference) strains.
Mapping results were obtained with MAPMAKERIQTL Version 1.1, using an unconstrained model. A black triangle at the bottom indicates peak LOD score at M134G01 marker. Horizontal line at the bottom shows a 1-LOD confidence interval.
Fig. 1B shows SAC1-containing chromosomal region defined by a donor fragment of the 129.B6-Sacb partially congenic mice. The partially congenic strains were constructed by identifying several founder F2 mice with small fragments of the telomeric region of mouse chromosome 4 from the B6 strain and successive backcrossing to the 129 strain. Presence and size of donor fragment were determined by genotyping polymorphic markers in mice from the N4, N6, N7, N4F4, and N3F5 generations.
Fig. 1 C shows average daily saccharin consumption by N6, N7, N4F4, and N3F5 segregating partially congenic 129.B6-Sac mice in 4-days two-bottle tests with water (means ~ SE). The open bar indicates intakes of mice that did not inherit the donor fragment. The black bar indicates intakes of mice with one or two copies of the donor fragment, which is flanked by 280612-T7 proximally and D4Mon1 distally. The complete donor fragment is represented by overlapping sequences of the BAC RPCI-23-118E21 and a genomic clone (Accession AF 185591 ), as indicated at the bottom. The size of the SAC 1-containing donor fragment is 194, 478 kb.
Fig. 1D shows BAC contig of distal chromosome 4 in the SACl region.
Using 32P radioactively labeled probes from the nonrecombinant interval, a mouse BAC library (RPCI-23) was screened; positive clones were confirmed by PCR analysis and only clones positive by hybridization and by PCR are included in the contig. BAC ends were sequenced and PCR primers designed. The STS
content of each BAC, using all BAC ends was determined. BAC size was determined by digesting the BAC with Notl, and the insert size determined using pulse field gel electrophoresis.
Fig. 1E shows genes contained within the SACl nonrecombinant interval.
Arrows indicate predicted direction of transcription. See Table 1 for a description of gene prediction, and details concerning function.
Fig. 2A shows the mouse SAC1 gene (mSac; Accession AF311386), its human ortholog (hSac), and the previously described gene TlRl, now Gpr70, are aligned above. Residues shaded in black are identical between at least two identical residues; residues in gray indicate conservative changes. The human ortholog was identified by sequence homology search within the htgs database (Accession AC026283). The amino acid sequence of the human ortholog was predicted using GENSCAN. The amino acid sequence of mouse Gpr70 was obtained by constructing primers based upon the nucleotide sequence, and taste cDNA was amplified and sequenced. This amino acid and nucleotide sequence for Gpr70 differed slightly from the initial report; the sequence reported in this paper has been deposited in GenBank (AF301161, AF301162). The location of the missense mutation is indicated by an *.
Fig. 2B shows structure of the SAC1 gene. The six exons are shown as black boxes.
Fig. 2C shows conformation of a protein predicted from the Sac gene. To determine the transmembrane regions, the hydrophobicity was determined using the computer program HMMTOP, and drawn with TOPO. The missense mutation is denoted with an asterisk.
Fig. 3 shows saccharin and sucrose preferences by mice from inbred strains with two different haplotypes of the Sac gene. The haplotype found in the B6 mice and the other high sweetener-preferring inbred strains consisted of four variants, two variants were 5' of the predicted translation start codon, one variant was a missense mutation (I1e61 Thr), and the last variant was located in the intron between exon 2 and 3. The strains with the B6-like haplotype of Sac strongly preferred saccharin (82 ~ 4%) and sucrose (86 ~ 6%), whereas strains with the 129-like haplotype were indifferent to these solutions (57 ~ 2% and 54 ~ 1 respectively, p = 0.0015).
Fig. 4A shows tissue expression of the SAC 1 gene. Note that cDNA was obtained from a commercial source for the multiple tissue panel, with the exception of tongue cDNA, which was as isolated by the investigator, as described within the text. Relative band intensities may differ due to differences in cDNA
isolation methods or concentration.
Fig. 4B shows RNA from human fungiform papillae was obtained from biopsy material, reversed transcribed, and the resulting bands from genomic and cDNA were amplified using primers, described in the text. The bands were excised from the agarose gel, purified and reamplified. The PCR product was sequenced to confirm that the bands amplified the human otholog to Sac.
Fig. 5 shows amino acid sequence alignment of the mouse cDNA sequence for the SAC 1 gene and the cDNA for a calcium sensing metabotropic receptor.
Dark areas indicated regions of shared similarity.
Fig. 6 plots the hydrophobicity of the SACl amino acid sequence as predicted by the computer program Top Pred. Note the seven transmembrane domains characteristic of G-protein coupled receptors.

DETAILED DESCRIPTION OF THE INVENTION
I. Definitions The present invention employs the following definitions:
As used herein, the terms "polynucleotide" and "nucleic acid" refer to naturally occurring polynucleotides, e.g., DNA or RNA. These terms do not refer to a specific length. Thus, these terms include oligonucleotide, primer, probe, etc.
These terms also refer to analogs of naturally occurring polynucleotides. The polynucleotide may be double stranded or single stranded. The polynucleotides may be labeled with radiolabels, fluorescent labels, enzymatic labels, proteins, haptens, antibodies, sequence tags.
For example, these terms include RNA, cDNA, genomic DNA, synthetic forms, and mixed polymers, both sense and antisense strands, and may be chemically or biochemically modified or may contain non-natural or derivatized nucleotide bases, as will be readily appreciated by those skilled in the art.
Such modifications include, for example, labels, methylation, substitution of one or more of the naturally occurring nucleotides with an analog, internucleotide modifications such as uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoramidates, carbamates, etc.), charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), pendent moieties (e.g., polypeptides), intercalators (e.g., acridine, psoralen, etc.), chelators, alkylators, and modified linkages (e.g., alpha anomeric nucleic acids, etc.). Also included are synthetic molecules that mimic polynucleotides in their ability to bind to a designated sequence via hydrogen bonding and other chemical interactions. Such molecules are known in the art and include, for example, those in which peptide linkages substitute for phosphate linkages in the backbone of the molecule.
As used herein, the term "polynucleotide amplification" refers to a broad range of techniques for increasing the number of copies of specific polynucleotide sequences. Typically, amplification of either or both strand of the target nucleic acid comprises the use of one or more nucleic acid-modifying enzymes, such as a DNA polymerase, a ligase, an RNA polymerase, or an RNA-dependent reverse transcriptase. Examples of polynucleotide amplification reaction include, but not _7_ limited to, polymerase chain reaction (PCR), nucleic acid sequence based amplification (NASB), self sustained sequence replication (3SR), strand displacement activation (SDA), ligase chain reaction (LCR), Q[3 replicase system, and the like.
As used herein, the term "primer" refers to a nucleic acid, e.g., synthetic polynucleotide, which is capable of annealing to a complementary template nucleic acid (e.g., the SAC1 locus) and serving as a point of initiation for template-directed nucleic acid synthesis. A primer need not reflect the exact sequence of the template but must be sufficiently complementary to hybridize with a template. Typically, a primer will include a free hydroxyl group at the 3' end. The appropriate length of a primer depends on the intended use of the primer but typically ranges from 12 to 30 nucleotides. The term primer pair means a set of primers including a 5' upstream primer that hybridizes with the 5° end of the target sequence to be amplified and a 3' downstream primer that hybridizes with the complement of the 3' end of the target sequence to be amplified.
The present invention includes all novel primers having at least eight nucleotides derived from the SAC1 locus for amplifying the SACl gene, its complement or functionally equivalent nucleic acid sequences. The present invention does not include primers which exist in the prior art. That is, the present invention includes all primers having at least 8 nucleotides with the proviso that it does not include primers existing in the prior art.
"Target polynucleotide" refers to a single- or double-stranded polynucleotide which is suspected of containing a target sequence, and which may be present in a variety of types of samples, including biological samples.
"Antibody" refers to polyclonal and/or monoclonal antibody and fragments thereof, and.immunologic binding equivalents thereof, which are capable of specifically binding to the SAC 1 polypeptides and fragments thereof or to polynucleotide sequences from the SAC 1 region, particularly from the SAC1 locus or a portion thereof. Antibody may be a homogeneous molecular entity, or a mixture such as a serum product made up of a plurality of different molecular entities.

_g_ Antibodies may be produced by in vitro or in vivo techniques well-known in the art. For example, for production of polyclonal antibodies, an appropriate target immune system, typically mouse or rabbit, is selected. Substantially purified antigen is presented to the immune system. Typical sites for injection are in footpads, intramuscularly, intraperitoneally, or intradermally. Polyclonal antibodies may then be purified and tested for immunological response, e.g., using an immunoassay.
For production of monoclonal antibodies, protein, polypeptide, fusion protein, or fragments thereof may be injected into mice. After the appropriate period of time, the spleens may be excised and individual spleen cells fused, typically, to immortalized myeloma cells under appropriate selection conditions.
Thereafter, the cells are clonally separated and the supernatants of each clone tested for their production of an appropriate antibody specific for the desired region of the antigen. Affinities of monoclonal antibodies are typically 10-g or preferably 10-9 to 10-10 M-1 or stronger.
Other suitable techniques involve in vitro exposure of lymphocytes to the ° antigenic polypeptides, or alternatively, to selection of libraries of antibodies in phage or similar vectors.
Frequently, antibodies are labeled by joining, either covalently or non-covalently, a substance which provides for a detectable signal. A wide variety of labels and conjugation techniques are known. Suitable labels include radionuclides, enzymes, substrates, cofactors, inhibitors, fluorescent agents, chemiluminescent agents, magnetic particles, and the like. Also, recombinant immunoglobulins may be produced.
"Binding partner" refers to a molecule capable of binding another molecule with specificity, as for example, an antigen and an antigen-specific antibody or an enzyme and its inhibitor. Binding partners are known in the art and include, for example, biotin and avidin or streptavidin, IgG and protein A, receptor-ligand couples, and complementary polynucleotide strands. In the case of complementary polynucleotide binding partners, the partners are normally at least about 15, 20, 25, 30, 40 bases in length.

A "biological sample" refers to a sample of tissue or fluid suspected of containing an analyte (e.g., polynucleotide, polypeptide) including, but not limited to, e.g., plasma, serum, spinal fluid, lymph fluid, the external sections of the skin, respiratory, intestinal, and genitourinary tracts, tears, saliva, blood cells, organs, tissue and samples of in vitro cell culture constituents. A biological sample is typically from human or other animal.
"Encode." A polynucleotide is said to "encode" a polypeptide if, in its native state or when manipulated by methods well-known to those skilled in the art, it can be transcribed and/or translated to produce the mRNA and/or the polypeptide .or a fragment thereof. The antisense strand is the complement of such a nucleic acid, and the encoding sequence can be deduced therefrom.
"Isolated" or "substantially pure" polynucleotide or polypeptide (e.g., an RNA, DNA, protein) is one which is substantially separated from other cellular components which naturally accompany a native human nucleic acid or protein, e.g., ribosomes, polymerases, many other human genome sequences and proteins.
The term embraces a nucleic acid or peptide sequence which has been removed from its naturally occurring environment, and includes recombinant or cloned DNA isolates and chemically synthesized analogs or analogs biologically synthesized by heterologous systems.
"SAC 1 Allele" refers to normal alleles of the SAC 1 locus as well as alleles carrying variations that predispose individuals to develop obesity, diabetes, or for alcohol consumption or alcoholism.
"SAC1 Locus" refers to polynucleotides, which are in the SAC1 region, that are likely to be expressed in normal individual, certain alleles of which predispose an individual to develop obesity, diabetes, or alcohol consumption or alcoholism. The SACl locus includes coding sequences, intervening sequences and regulatory elements controlling transcription and/or translation. The SAC1 locus includes all allelic variations of the DNA sequence.
The DNA sequences used in this invention will usually comprise at least about S codons (15 nucleotides), 7, 10, 15, 20, or 30 codons, and most preferably, at least about 35 codons. One or more introns may also be present. This number of nucleotides is usually about the minimal length required for a successful probe that would hybridize specifically with a SAC 1 locus.
"SAC1 Region" refers to a portion of mouse chromosome 4 bounded by the markers 280612-T7 and D4Monl GenBank Accession number is YG7772 (SEQ ID NO: 652) and is GCAGTGAGCTGCAGAGTTTGCAGAATGAGGGCACTCTAAACTCATCAA
GTGAGGAGGCCCTTCCCTCACACTCCAGATGGCTGATAGGTGGCATTA
CATGGTC(CA)nCGCGCGCACGCGCTCAGATGCAATCTCCACATTCATA
ACCAGATGTCCTTGGGTAGGCCT. The CA sequence in the middle is variable in length. In the B6 mouse, n =19, while in the 129 mouse, n =16.
This region contains the SAC1 locus, including the SAC1 gene. GenBank accession number for the SAC1 gene is AF311386.
As used herein, a "portion" or "fragment" of the SACl gene, locus, region, or allele is defined as having a minimal size of at least about 15 nucleotides, or preferably at least about 20, or more preferably at least about 25 nucleotides, and may have a minimal size of at least about 40 nucleotides.
As used herein, the term "polypeptide" refers to a polymer of amino acids without referring to a specific length. This term includes to naturally occurring protein. The term also refers to modifications, analogues and functional mimetics thereof. For example, modifications of the polypeptide may include glycosylations, acetylations, phosphorylations, and the like. Analogues of polypeptide include unnatural amino acid, substituted linkage; etc. Also included are polypeptides encoded by DNA which hybridize under high or low stringency conditions, to the nucleic acids of interest.
Modification of polypeptides includes those substantially homologous to primary structural sequence, e.g., in vivo or in vitro chemical and biochemical modifications or incorporation unusual amino acids. Such modifications include, for example, acetylation, carboxylation, phosphorylation, glycosylation, ubiquitination, labeling, e.g., with radionuclides, and various enzymatic modifications, as will be readily appreciated by those well-skilled in the art. A
variety of methods for labeling polypeptides and of substituents or labels useful for such purposes are well-known in the art, and include radioactive isotopes such as 32P, ligands which bind to labeled antiligands (e.g., antibodies), fluorophores, chemiluminescent agents, enzymes, and antiligands which can serve as specific ' binding pair members for a labeled ligand. The choice of label depends on the sensitivity required, ease of conjugation with the primer, stability requirements, and available instrumentation. Methods of labeling polypeptides are well-known in the art (see Sambrook et al., 1989 or Ausubel et al., 1992).
Besides substantially full-length polypeptides, the present invention provides for biologically active fragments of the polypeptides. Significant biological activities include ligand-binding, immunological activity, and other biological activities characteristic of SAC1 polypeptides. Immunological activities include both immunogenic function in a target immune system, as well as sharing of immunological epitopes for binding, serving as either a competitor or substitute antigen for an epitope of the SAC1 protein. As used herein, "epitope" refers to an antigenic determinant of a polypeptide. An epitope could comprise three amino acids in a spatial conformation that is unique to the epitope. Generally, an epitope consists of at least five such amino acids, and more usually consists of at least 8 to 10 such amino acids. Methods of determining the spatial conformation of such amino acids are known in the art.
For immunological purposes, tandem-repeat polypeptide segments may be used as immunogens, thereby producing highly antigenic proteins.
Alternatively, such polypeptides will serve as highly efficient competitors for specific binding.
Fusion proteins comprise SACl polypeptides and fragments. Homologous polypeptides may be fusions between two or more SAC 1 polypeptide sequences or between the sequences of SACl and a related protein. Likewise, heterologous fusions may be constructed which would exhibit a combination of properties or activities of the derivative proteins. For example, ligand-binding or other domains may be "swapped" between different new fusion polypeptides or fragments. Such homologous or heterologous fusion polypeptides may display, for example, altered strength or specificity of binding. Fusion partners include immunoglobulins, bacterial ~i-galactosidase, trpE, protein A, (3-lactamase, oc-amylase, alcohol dehydrogenase, and yeast a mating factor.

Fusion proteins will typically be made by either recombinant nucleic acid methods or may be chemically synthesized. Techniques for the synthesis of polypeptides are known in the art.
Functional mimetics of a native polypeptide may be obtained using known methods in the art. For example, polypeptides may be least about 50%
homologous to the native amino acid sequence, preferably in excess of about 70%, and more preferably at least about 90% homologous. Substitutions typically contain the exchange of one amino acid for another at one or more sites within the polypeptide, and may be designed to modulate one or more properties of the polypeptide, such as stability against proteolytic cleavage, without the loss of other functions or properties. Amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues involved. Preferred substitutions are ones which are conservative, that is, one amino acid is replaced with one of similar shape and charge. Conservative substitutions are well-known in the art and typically include substitutions within the following groups: glycine, alanine;
valine, isoleucine, leucine; aspartic acid, glutamic acid; asparagine, glutamine;
serine, threonine; lysine, arginine; and tyrosine, phenylalanine.
Certain amino acids may be substituted for other amino acids in a polypeptide structure without appreciable loss of interactive binding capacity with structures such as, for example, antigen-binding regions of antibodies or binding sites on substrate molecules or binding sites on proteins interacting with a polypeptide. Since it is the interactive capacity and nature of a polypeptide which defines that polypeptide's biological functional activity, certain amino acid substitutions can be made in a protein sequence, and its underlying DNA coding sequence, and nevertheless obtain a protein with like properties. In making such changes, the hydropathic index of amino acids may be considered. The importance of the hydrophobic amino acid index in confernng interactive biological function on a protein is generally understood in the art.
Alternatively, the substitution of like amino acids can be made effectively on the basis of hydrophilicity.

A peptide mimetic may be a peptide-containing molecule that mimics elements of protein secondary structure. The underlying rationale behind the use of peptide mimetics is that the peptide backbone of proteins exists chiefly to orient amino acid side chains in such a way as to facilitate molecular interactions, such as those of antibody and antigen, enzyme and substrate or scaffolding proteins. A
peptide mimetic is designed to permit molecular interactions similar to the natural molecule. A mimetic may not be a peptide at all, but it will retain the essential biological activity of a natural polypeptide.
Polypeptides may be produced by expression in a prokaryotic cell or produced synthetically. These polypeptides typically lack native post-translational processing, such as glycosylation. Polypeptides may be labeled with radiolabels, fluorescent labels, enzymatic labels, proteins, haptens, antibodies, sequence tags.
"SACl polypeptide" refers to a protein or polypeptide encoded by the SAC1 locus, variants, fragments or functional mimics thereof. A SAC
polypeptide may be that derived from any of the exons described herein which may be in isolated and/or purified form. The length of SAC1 polypeptide sequences is generally at least about S amino acids, usually at least about 10, 15, 20, 30 residues.
"Alcohol consumption" relates to the intake andlor preference of an animal for ethanol.
"Diabetes" refers to any disorder that exhibits phenotypic features of an increased or decreased level of a biological substance associated with glucose or fatty acid metabolism. The term "carbohydrate" refers to simple mono and disaccharides.
The terms "sequence variation" or "variant form" encompass all forms of polymorphism and mutations. A sequence variation may range from a single nucleotide variation to the insertion, modification, or deletion of more than one nucleotide. A sequence variation may be located at the exon, intron, or regulatory region of a gene.
Polymorphism refers to the occurrence of two or more genetically determined alternative sequences or alleles in a population. A biallelic polymorphism has two forms. A triallelic polymorphism has three forms. A

polymorphic site is the locus at which sequence divergence occurs. Diploid organisms may be homozygous or heterozygous for allelic forms. Polymorphic sites have at least two alleles, each occurring at frequency of greater than 1 % of a selected population. Polymorphic sites also include restriction fragment length polymorphisms, variable number of tandem repeats (VNTRs), hypervariable regions, minisatellites, dinucleotide repeats, trinucleotide repeats, tetranucleotide repeats, simple sequence repeats, and insertion elements. The first identified allelic form may be arbitrarily designated as the reference sequence and other allelic forms may be designated as alternative or variant alleles. The allelic form occurring most frequently in a selected population is sometimes referred to as the wild type form or the consensus sequence.
Mutations include deletions, insertions and point mutations in the coding and noncoding regions. Deletions may be of the entire gene or of only a portion of the gene. Point mutations may result in stop codons, frameshift mutations, or amino acid substitutions.. Somatic mutations are those which occur only in certain tissues, such as liver, heart, etc. and are not inherited in the germline.
Germline mutations can be found in any of a body's tissues and are inherited.
"Operably linked" refers to a juxtaposition wherein the components are in a relationship permitting them to function in their intended manner. For instance, a promoter is operably linked to a coding sequence if the promoter affects its transcription or expression.
The term "probes" refers to polynucleotide of any suitable length which allows specific hybridization to the target region. Probes may be attached to a label or reporter molecule using known methods in the art. Probes may be selected by using homologous polynucleotides. Alternatively, polynucleotides encoding these or similar polypeptides may be synthesized or selected by use of the redundancy in the genetic code. Various codon substitutions may be introduced, e.g., by silent changes (thereby producing various restriction sites) or to optimize expression for a particular system. Mutations may be introduced to modify the properties of the polypeptide, perhaps to change ligand-binding affinities, interchain affinities, or the polypeptide degradation or turnover rate.

Probes comprising synthetic oligonucleotides or other polynucleotides of the present invention may be derived from naturally occurring or recombinant single- or double-stranded polynucleotides, or be chemically synthesized.
Probes may also be labeled by nick translation, Klenow fill-in reaction, or other methods known in the art.
Portions of the polynucleotide sequence having at least about 8 nucleotides, usually at least about 1 S nucleotides, and fewer than about 6 kb, usually fewer than about 1.0 kb, from a polynucleotide sequence encoding SAC1 are preferred as probes.
The terms "isolated," "substantially pure," and "substantially homogeneous" are used interchangeably to describe a protein or polypeptide which has been separated from components which accompany it in its natural state. A monorneric protein is substantially pure when at least about 60% to 7S%
of a sample exhibits a single polypeptide sequence. A substantially pure protein 1 S will typically comprise about 60% to 90% W/W of a protein sample, more usually about 9S%, and preferably will be over about 99% pure. Protein purity or homogeneity may be indicated by a number of means well-known in the art, such as polyacrylamide gel electrophoresis of a protein sample, followed by visualizing a single polypeptide band upon staining the gel. For certain purposes, higher resolution may be provided by using HPLC or other means well-known in the art which are utilized for purification.
A SACl protein is substantially free of naturally associated components when it is separated from the native contaminants which accompany it in its natural state. Thus, a polypeptide which is chemically synthesized or synthesized 2S in a cellular system different from the cell from which it naturally originates will be substantially free from its naturally associated components. A protein may also be rendered substantially free of naturally associated components by isolation, using protein purification techniques well-known in the art.
"Recombinant nucleic acid" is a nucleic acid which is not naturally occurnng, or which is made by the artificial combination of two otherwise separated segments of sequence. This artificial combination is often accomplished by either chemical synthesis means, or by the artificial manipulation of isolated segments of nucleic acids, e.g., by genetic engineering techniques. Such is usually done to replace a codon with a redundant codon encoding the same or a conservative amino acid, while typically introducing or removing a sequence recognition site. Alternatively, it is performed to join together nucleic acid segments of desired functions to generate a desired combination of functions.
"Regulatory sequences" refers to those sequences normally within 100 kb of the coding region of a locus, but they may also be more distant from the coding region, which affect the expression of the gene (including transcription of the gene, and translation, splicing, stability or the like of the messenger RNA).
"Substantial homology or similarity." A nucleic acid or fragment thereof is of substantially homologous ("or substantially similar") to another if, when optimally aligned (with appropriate nucleotide insertions or deletions) with the other nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 60% of the nucleotide bases, usually at least about 70%, more usually at least about 80%, preferably at least about 90%, and more preferably at least about 95-98% of the nucleotide bases.
Identity means the degree of sequence relatedness between two polypeptide or two polynucleotides sequences as determined by the identity of the match between two strings of such sequences. Identity can be readily calculated (Lesk A.M., ed., Computational Molecular Biology, New York: Oxford University Press, 1988; Smith D.W., ed., Biocomputing: hzformatics and Gen, ome Projects, New York: Academic Press, New York, 1993; Griffin A.M., and Griffin H.G., eds., Computer Analysis of Sequence Data, Part l, New Jersey:
Humana Press, 1994; von Heinje G., Sequence Analysis in Molecular Biology, Academic Press, 1987; and Gribskov M. and Devereux J., eds., Sequence Analysis Primer, New York: M Stockton Press, 1991).
Alternatively, substantial homology or similarity exists when a nucleic acid or fragment thereof will hybridize to another nucleic acid (or a complementary strand thereof) under selective hybridization conditions, to a strand, or to its complement. Selectivity of hybridization exists when hybridization which is substantially more selective than total lack of specificity occurs. Typically, selective hybridization will occur when there is at least about Glu Leu Val Met Ala Leu A

55% homology over a stretch of at least about 14 nucleotides, preferably at least about 65%, more preferably at least about 75%, and most preferably at least about 90%. The length of homology comparison, as described, may be over longer stretches, and in certain embodiments will often be over a stretch of at least about 9 nucleotides, usually at least about 20 nucleotides, more usually at least about 24 nucleotides, typically at least about 28 nucleotides, more typically at least about 32 nucleotides, and preferably at least about 36 or more nucleotides.
Nucleic acid hybridization will be affected by such conditions as salt concentration, temperature, or organic solvents, in addition to the base composition, length of the complementary strands, and the number of nucleotide base mismatches between the hybridizing nucleic acids, as will be readily appreciated by those skilled in the art. Stringent temperature conditions will generally include temperatures in excess of 30°C, typically in excess of 37°C, and preferably in excess of 45°C. Stringent salt conditions will ordinarily be less than 1000 mM, typically less than 500 mM, and preferably less than 200 mM.
However, the combination of parameters is much more important than the measure of any single parameter.
The terms "substantial homology" or "substantial identity," when referring to polypeptides, indicate that the polypeptide or protein in question exhibits at least about 30% identity with an entire naturally-occurring protein or a portion thereof, usually at least about 70% identity, and preferably at least about 95%
identity.
Homology, for polypeptides, is typically measured using sequence analysis software (see, e.g., the Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center). Protein analysis software matches similar sequences using measures of homology assigned to various substitutions, deletions and other modifications.
Conservative substitutions typically include substitutions within the following groups:
glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid;
asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine.
"Substantially similar function" refers to the function of a modified nucleic acid or a modified protein, with reference to the wild-type SAC1 nucleic acid or wild-type SAC1 polypeptide. The modified polypeptide will be substantially homologous to the wild-type SAC1 polypeptide and will have substantially the same function. The modified polypeptide may have an altered amino acid sequence and/or may contain modified amino acids. In addition to the similarity of function, the modified polypeptide may have other useful properties, such as a longer half life. The similarity of function (activity) of the modified polypeptide may be substantially the same as the activity of the wild-type SACl polypeptide.
Alternatively, the similarity of function (activity) of the modified polypeptide may be higher than the activity of the wild-type SAC 1 polypeptide. The modified polypeptide is synthesized using conventional techniques, or is encoded by a modified nucleic acid and produced using conventional techniques. The modified nucleic acid is prepared by conventional techniques. A nucleic acid with a function substantially similar to the wild-type SAC1 gene function produces the modified protein described above.
A polypeptide "fragment," "portion," or "segment" is a stretch of amino acid residues of at least about 5 to 7 contiguous amino acids, often at least about 7 to 9 contiguous amino acids, typically at least about 9 to 13 contiguous amino acids and, most preferably, at least about 20 to 30 or more contiguous amino acids.
The polypeptides of the present invention, if soluble, may be coupled to a solid-phase support, e.g., nitrocellulose, nylon, column packing materials (e.g., Sepharose beads), magnetic beads, glass wool, plastic, metal, polymer gels, cells, or other substrates. Such supports may take the form, for example, of beads, wells, dipsticks, or membranes.
"Target region" refers to a region of the nucleic acid which is amplified and/or detected. The term "target sequence" refers to a sequence with which a probe or primer will form a stable hybrid under desired conditions.
II. Positional Cloning of Mouse SACl Gene and the Discovery of a Gene and Its Sequence Variation Associated With Altered Sensation for Carbohydrates Inbred strains of mice differ in their intake of sweeteners (Bachmanov A.A., Reed D.R., Tordoff M.G., Price R.A., and Beauchamp G.K. Intake of ethanol, sodium chloride, sucrose, citric acid, and quinine hydrochloride solutions by mice: a genetic analysis. Behavior Genetics, 1996;26:563-573; Lush LE., .The S genetics of tasting in mice. VI. Saccharin, acesulfame, dulcin and sucrose.
Genet Res, 1989;53:95-99; Lush I. The genetics of bitterness, sweetness, and saltiness in strains of mice. In Genetics of Perception and Communication, Vol. 3, eds.
Wysocki C. and Kare M., New York: Marcel Dekker, 1991:227-235;
Capretta P.J. Saccharin and saccharin-glucose ingestion in two inbred strains of Mus musculus. Psychon. Sci., 1970;21:133-135; Nachman M. The inheritance of saccharin preference. Journal of Comp Physiol Psychol, 1959;52:451-457).
Breeding and linkage experiments suggest that a single gene, the Sac locus (for saccharin intake), accounts for a large proportion of the genetic variance (Fuller J.L. Single-locus control of saccharin preference in mice. Journal of Heredity, 1974;65:33-36; Capeless C.G. and Whitney G. The genetic basis of preference for sweet substances among inbred strains of mice: preference ratio phenotypes and the alleles of the Sac and dpa loci. Chem Senses, 1995;20:291=
298; Bachmanov A.A. et al. Sucrose consumption in mice: major influence of two genetic loci affecting peripheral sensory responses. Mammalian Genome, 1997;8:545-548; Belknap J.K. et al. Single-locus control of saccharin intake in BXD/Ty recombinant inbred (RI) mice: some methodological implications for RI
strain analysis. Behav Genet, 1992;22:81-100; Blizard D.A., Kotlus B., and Frank M.E. Quantitative trait loci associated with short-term intake of sucrose, saccharin and quinine solutions in laboratory mice. Chem Senses, 1999;24:373-85). Using genetic and physical mapping methods, an interval of 194 kb was identified at the telomeric end of mouse chromosome 4 that contains the Sac locus. BAC sequencing within this interval led to the identification of a gene that has a 30% amino acid homology with other putative taste receptors (Noon M.A.
et al. Putative mammalian taste receptors: a class of taste-specific GPCRs with distinct topographic selectivity. Cell, 1999;96:541-551). This gene is expressed in mouse tongue. Mutation detection on this gene revealed a missense mutation (Ile6lThr) with four other sequence variants define a haplotype found in mice with low sweetener preference (129, Balb/c, AKR, and DBA2). An alternative five variant haplotype is found in mice with a high preference for sweet fluids (B6, SWR, IS, ST, and SEA). A human ortholog of this gene exists, and is expressed in human taste papillae. We therefore suggest that this gene is a sweet taste receptor, and variation within this gene is responsible for the phenotype of the Sac locus.
To identify this locus, mice from the high sweetener preference (C57BLl6ByJ; B6) and the low sweetener preference (129P3/J; formerly 129/J, abbreviated here as 129) were used as parental strains to produce an F2 generation. The F2 mice were phenotyped for sweetener preference using 96-hour two-bottle taste tests and genotyped with markers polymorphic between the B6 and 129 strains (Fig. 1A). The results of this analysis indicated peak linkage near marker D18346 with the B6 allele having a dominant mode of inheritance.
Using recombinant mice from the F2 generation, 129.B6-Sac partially congenic mice were created, using genotypic (B6 allele at D18346; Fig. 1B) and phenotypic (high saccharin intake; Fig. 1 C) characteristics as selection criteria for each generation. Genotyping of partially congenic mice with polymorphic markers defined the Sac nonrecombinant interval. Radiation hybrid mapping was conducted with additional markers (R74924, D18402, D18346, Agrin, V2r2 and D4Ertd296e). These markers were amplified using DNA and mouse and hamster control DNA in the T31 mouse radiation hybrid panel, scored for the presence or absence of an appropriately sized band, and the data analyzed by the Jackson Laboratory. All markers were within the SAC1 confidence interval suggested by the initial linkage analysis, and were used in subsequent analyses.
A BAC library was screened with markers within the nonrecombinant interval, and a contig was developed (Fig. 1D). A BAC clone was selected for sequencing (RPCI-23-118E21, 246 kb). Within this BAC, a gene with a 30%
homology to T1R1 (a putative taste receptor) was discovered (Fig. 2A), along with other ESTs and known genes (Table 1). The human orthalog to this gene was identified from a BAC available in the public htgs database, and the predicted protein sequence was aligned with SACl and TlRl. SACl is 858 amino acids in length and contains six exons; the intron and exon boundaries were determined by sequencing of the mouse tongue cDNA (Fig. 2B). The secondary structure of this protein with regards to transmembrane domains was predicted (Fig. 2C).
To determine whether this gene might contain functional polymorphisms that could account for the behavioral differences between the two strains, 11.8 kb of sequence, including the SACl gene and several kb up and downstream were amplified with PCR primers and then sequenced using DNA from the high and low prefernng strains (Lush LE., The genetics of tasting in mice. VI.
Saccharin, acesulfame, dulcin and sucrose. Genet Res1989;53:95-99; Lush I. The genetics of bitterness, sweetness, and saltiness in strains of mice. In Genetics of Perception and Communication, Vol. 3, eds. Wysocki C. and Kare M., New York: Marcel Dekker, 1991:227-235). Many variants existed between these strains, and of these, five variants were found in the low preferring strains but not in the high preferring strain. One of these variants results in a missense mutation (Ile6lThr; Fig.
2). The other four variants were in non-coding regions (T>A -2383 nt; A>G -183 nt;
A>G +134 nt; 'I>C +651 nt, between exon 2 and 3). These five variants will be referred to as the 129-like or B6-like haplotypes. Additional inbred strains of mice with known saccharin and sucrose preferences (Lush LE., The genetics of tasting in mice. VI. Saccharin, acesulfame, dulcin and sucrose. Genet Res, 1989;53:95-99; Lush I. The genetics of bitterness, sweetness, and saltiness in strains of mice. In Genetics ofPerception and Communication, Vol. 3, eds.
Wysocki C. and Kare M., New York: Marcel Dekker, 1991:227-235; Lush LE.
and Holland G. The genetics of tasting in mice. V. Glycine and cyclohexamide.
Genet Res, 1988;52:207-212) were also sequenced. The 129-like haplotype was found in mice with lower sweetener preference and the B6-like haplotype was found in mice with higher sweetener preference (Fig. 3).
B6 mice have higher maximal gustatory neural firing in response to sweeteners compared with 129 mice, as do the 129.B6-Sac partially congenic strains (Bachmanov A.A. et al. Sucrose consumption in mice: major influence of two genetic loci affecting peripheral sensory responses. Mammalian Genome, 1997;8:545-548). Thus, the SAC1 gene is likely to be expressed in tongue. To test this hypothesis, RNA from mouse and human tongue was extracted, reversed transcribed into cDNA and primers, chosen to span an intron, were used in a PCR

reaction. Genomic and cDNA yielded bands of different sizes, which were purified and sequenced (Figure 4AB). Sequencing results confirmed that the bands were derived from this gene with the appropriate intron/exon boundaries.
Further analysis of expression in cDNA in mouse tissue, using commercially available mouse cDNA, indicated this gene is also expressed is widely expressed.
The broad range of tissue expression of this gene may indicate that other tissues use this receptor to sense extra cellular sugars (Fig. 4A).
Hoon et al. identified a gene, Gpr70 (formerly TRl or T1R1) as a putative sweet receptor based mainly on its expression in anterior tongue taste cells.
Since it also mapped to distal chromosome 4, it was a logical candidate for SAC 1.
However, we have shown that Gpr70 is at least 4 cM proximal to SAC1 (Li X.
et al. The saccharin preference locus (Sac) and the putative sweet taste receptor (Gpr70) gene have distinct locations on mouse chromosome 4. Mammalian Genome, 2001;12:13-16). Nevertheless, Gpr70 could be an additional sweet receptor and there could be others. It has been argued based upon human psychophysical studies and studies of sweet taste transduction mechanisms that there must be more than one sweet receptor. Other lines of evidence, however, are more consistent with the existence of one or a very few receptors (Bartoshuk L.M.
Is sweetness unitary? An evaluation of the evidence for multiple sweeteners.
In Sweetness, ed. bobbing, J., London: Springer-Verlag, 197:33-46). At present no evidence has been found of a family of Sac-like receptors resembling the large family of bitter receptors recently reported (Matsunami H., Montmayeur J.P., and Buck L.B. A family of candidate taste receptors in human and mouse [see comments]. Nature, 2000;404:601-604; Adler E. et al. A novel family of mammalian taste receptors [see comments]. Cell, 2000;100:693-702). The sweet substances that exist in nature, which presumably shaped the evolution of sweet receptor(s), are likely much more similar amongst themselves, mostly simple sugars, than are the vast array of structurally diverse bitter tasting compounds.
A receptor for the sugar trehalose has recently been identified in the fruit fly, Drosophila melanogaster. Surprisingly, the trehalose and other fly taste receptors, have no homology with SAC 1. The specialization of flies for the sugar trehalose may account for this divergence.

There may be multiple sweet receptors; evidence from across species comparisons, psychophysical cross adaptation, and sweetness competitors has been reviewed (Bartoshuk L.M. Is sweetness unitary? An evaluation of the evidence for multiple sweeteners. In Sweetness, ed. bobbing, J., London:
Springer-Verlag, 1987:33-46). The SAC 1 gene accounts for ~40% of the genetic differences in sweet perception between these two particular strains of mice, but other receptors, and other alleles of these receptors may exist.
Because sucrose is perceived to be bad for human health, considerable resources are directed toward the discovery of high potency, low caloric sweeteners. Most of the most widely known high potency sweeteners were discovered serendipitously, i.e., the sweetener was synthesized for a different purpose and someone in the laboratory accidentally tasted it and discovered it was sweet (Waiters E.D. The rational discovery of sweeteners. In Sweeteners.
Discovery, molecular design, and chemoreception, eds. Welters D.E., Orthoefer F.T., and DuBois G.E., American Chemical Society, USA, 1991:1-11).
More direct methods, however, have been employed to identify new sweet compounds, and the sweet receptor has been extensively modeled to predict which ligands will be sweet.
It is not known how or why different alleles of SAC1 arose in inbred strains of mice but their existence, in addition to providing us with a tool to identify a sweet receptor, raises the question of whether they might also characterize human populations. There appear to exist reliable individual differences in human sensitivity and preference for sweet sugars but whether these are genetically influenced remains to be determined. The identification of SAC

should facilitate research in this area. Also, the observation that SAC 1 is expressed in several tissues in addition to tongue raises the interesting possibility that it could be involved in other aspects of sugar recognition and that allelic variants in this gene could be related to diseases or conditions such as diabetes and obesity.
Alleles of the gene described in this application are likely to account for the SAC 1 behavioral and neurological phenotype for four reasons. First, the nonrecombinant region is small, less than 194 kb; this gene lies within this nonrecombinant interval and the peak of LOD score corresponds closely with the location of the gene. Second, of the genes contained within this region, no others are viable candidates for SAC 1. Third, this gene has sequence homology to other putative taste receptors, and is expressed in the tongue. Finally, a haplotype with a missense mutation is found in mice with low sweetener preference but not in mice with high sweetener preference. These data strongly suggest that mutations of this gene account for differences in the acceptance and preference for sweeteners attributed to the SAC1 locus.
Among the multiple mechanisms involved in regulation of ethanol intake, one of the least appreciated factors is the perception of its flavor (Nachman M., Larne C., Le Magnen J. The role of olfactory and orosensory factors in the alcohol preference of inbred strains of mice. Physiology Behavior, 1971;6:53-95).
Although individual variability in the perception of ethanol flavor by adults and children was described over 60 years ago (Richter C.P. Alcohol as a food.
Quart.
J. Studies Alcohol, 1941;1:650-62), the hypothesis that individual differences in alcohol chemosensory perception can affect alcohol intake did not receive due attention. As a result, the relationship between alcohol chemosensation and intake is not well-understood. Humans perceive ethanol flavor as a combination of components, including sweetness, bitterness, odor and irritation (burning sensation), which depend on ethanol concentration (Green B.G. The sensitivity of the tongue to ethanol. Ann. NY. Acad. Sci., 1987;510:315-7; Bartoshuk L.M., Conner E., Grubin D., Karrer T., Kochenbach K., Palsco M., et al. PROP
supertasters and the perception of ethyl alcohol. Chem. Se~tses, 1993.). Rats detect sweet (sucrose-like) and bitter (quinine-like) sensory components in ethanol (Kiefer S.W., Lawrence G.J. The sweet-bitter taste of alcohol: aversion generalization to various sweet-quinine mixtures in the rat. Chem. Senses, 1988;13:633-41; Kiefer S.W., Mahadevan R.S. The taste of alcohol for rats as revealed by aversion generalization tests. Chem. Senses, 1993;18:509-22) and probably perceive the other components detected by humans as well.
The relationship between ethanol and sweetener perception and consumption .has been studied the most and is supported by several lines of evidence:

(a) Electrophysiological recordings from gustatory nerves indicate that lingual application of ethanol activates sweetener-responsive neural fibers (Hellekant G., Danilova V., Roberts T., Ninomiya Y. The taste of ethanol in a primate model: I. Chorda tympani nerve response in Macaca mulatta.
Alcohol, 1997;14:473-84; Sako N., Yamamoto T. Electrophysiological and behavioral studies on taste effectiveness of alcohols in rats. Am. J.
Physiol., 1999;276:8388-96).
(b) Conditioned taste aversions generalize between ethanol and sucrose (Kiefer S.W., Lawrence G.J. The sweet-bitter taste of alcohol: aversion generalization to various sweet-quinine mixtures in the rat. Chem. Senses, 1988;13:633-41; Kiefer S.W., Mahadevan R.S. The taste of alcohol for rats as revealed by aversion generalization tests. Chem. Sepses, 1993;18:509-22; Lawrence G.J., Kiefer S.W. Generalization of specific taste aversions to alcohol in the rat. Chem. Senses, 1987;12:591-9; Blizard D.A., McClearn G.E. Association between ethanol and sucrose intake in the laboratory mouse: exploration via congenic strains and conditioned taste aversion. Alcohol. Clin. Exp. Res., 2000;24:253-8.), suggesting that ethanol and sucrose share the same taste property, most likely sweetness.
(c) Genetic associations between preferences for ethanol and sweeteners were found among some rat and mouse strains and within their segregating crosses (Overstreet D.H., Kampov-Polevoy A.B., Rezvani A.H., Murelle L., Halikas J.A., Janowsky D.S. Saccharin intake predicts ethanol intake in genetically heterogeneous rats as well as different rat strains. Alcohol.
Clin. Exp. Res., 1993;17:366-9; Sinclair J.D., Kampov-Polevoy A., Stewart R., Li T-K. Taste preferences in rat lines selected for low and high alcohol consumption. Alcohol, 1992;9:155-60; Stewart R.B., Russell R.N., Lumeng L., Li T-K., Murphy J.M. Consumptions of sweet, salty, sour, and bitter solutions by selectively bred alcohol-preferring and alcohol-nonpreferring lines of rats. Alcohol. Clih. Exp. Res., 1994;18:375-81;
Belknap J.K., Crabbe J.C., Young E.R. Voluntary consumption of alcohol in 15 inbred mouse strains. Psychopharmacol., 1993;112:503-10;

Bachmanov A.A., Reed D.R., Tordoff M.G., Price R.A., Beauchamp G.K.
Intake of ethanol, sodium chloride, sucrose, citric acid, and quinine hydrochloride solutions by mice: a genetic analysis. Behav. Genet., 1996;26:563-73; Bachmanov A.A., Tordoff M.G., Beauchamp G.K.
Ethanol consumption and taste preferences in C57BL/6ByJ and 1291) mice. Alcohol. Clin. Exp. Res., 1996;20:201-6), reviewed in (Kampov-Polevoy A.B., Garbutt J.C., Janowsky D.S. Association between preference for sweets and excessive alcohol intake: a review of animal and human studies. Alcohol. Alcohol., 1999;34:386-95; Overstreet D.H., Rezvani A.H., Parsian A. Behavioural features of alcohol-preferring rats:
focus on inbred strains. Alcohol. Alcohol., 1999;34:378-85); with some exceptions (Phillips T.J., Crabbe J.C., Metten P., Belknap J.K.
Localization of genes affecting alcohol drinking in mice. Alcohol. Clin.
Exp. Res., 1994;18:931-941; Parsian A., Overstreet D.H., Rezvani A.H.
Independent segregation of alcohol and saccharin intakes in F2 progeny from FH/ACI intercross (Abstract). Alcohol. Clin. Exp. Res., 2000;24(Supplement):5 8A)).
(d) Human studies show that alcoholics have a stronger liking of concentrated sucrose compared with nonalcoholics (Kampov-Polevoy A.B., Garbutt J.C., Davis C.E., Janowsky D.S. Preference for higher sugar concentrations and Tridimensional Personality Questionnaire scores in alcoholic and nonalcoholic men. Alcohol. Clin. Exp. Res., 1998;22:610-4;
Kampov-Polevoy A.B., Garbutt J.C., Janowsky D. Evidence of preference for a higher concentration sucrose solution in alcoholic men. American Journal of Psychiatry, 1997;154:269-70).
There are several possible mechanisms that could underlie the association between sweetener and ethanol responses:
(a) Common peripheral taste mechanisms, which may involve the interaction of ethanol with a peripheral sweet taste transduction. At least one such common peripheral mechanism is mediated by the Gpr98 gene (SAC1 locus) encoding a sweet taste receptor (as described below).

(b) Common brain mechanisms. The regulation of ingestive responses to ethanol and sweeteners may involve common opioidergic, serotonergic and dopaminergic brain neurotransmitter systems (Gosnell B.A., Majchrzak M.J. Centrally administered opioid peptides stimulate saccharin intake in nondeprived rats. Pharm. Biochem. Behav., 1989;33:805-10;
George S.R., Roldan L., Lui A., Naranjo C.A. Endogenous opioids are involved in the genetically determined high preference for ethanol consumption. Alcohol. Clin. Exp. Res., 1991;15:668-72; Hubell C.L., Marglin S.H., Spitalnic S.J., Abelson M.L., Wild K.D., Reid L.D.
Opioidergic, serotoninergic, and dopaminergic manipulations and rats' intake of a sweetened alcoholic beverage. Alcohol, 1991;8:355-67;
Pucilowski O., Rezvani A.H., Janowsky D.S. Suppression of alcohol and saccharin preference in rats by a novel Ca2+ channel inhibitor, Goe 5438.
Psychopharmacol., 1992;107:447-52). These mechanisms could be responsible for the emotional response to the pleasantness of ethanol or sweeteners, or the motivational mechanisms driving their intakes.
(c) Common signals related to the caloric value of ethanol and sugars (Gentry R.T., Dole V.P. Why does a sucrose choice reduce the consumption of alcohol in C57BL/6J mice? Life Sci., 1987;40:2191-4). Ethanol is metabolized in the body through some of the same pathways as carbohydrates and provides comparable energy. Thus, energy derived from carbohydrates and ethanol may have similar rewarding effects through the same hunger and satiety mechanisms.
(d) Incidental genetic linkage. Different genes affecting responses to ethanol and sweeteners may reside nearby on the same chromosome.
Ethanol consumption is a complex trait, depending on multiple mechanisms of its regulation and determined by multiple genes. A body of evidence suggests that ethanol consumption may depend on perception of its flavor, and that there is an association between perception and consumption of ethanol and sweet-tasting compounds. However, only a few genes have been identified as candidates affecting ethanol consumption.

The present invention provides that a gene, SAC1, is associated with the detection of a sensing of carbohydrates, other sweet compounds, and alcohols including ethanol. The sequence of the mouse SAC1 cDNA (SEQ ID NO: 1) is:
ATGCCAGCTTTGGCTATCATGGGTCTCAGCCTGGCTGCTTTCCTGGAGC
TTGGGATGGGGGCCTCTTTGTGTCTGTCACAGCAATTCAAGGCACAAG
GGGACTACATACTGGGCGGGCTATTTCCCCTGGGCTCAACCGAGGAGG
CCACTCTCAACCAGAGAACACAACCCAACAGCATCCCGTGCAACAGGT
TCTCACCCCTTGGTTTGTTCCTGGCCATGGCTATGAAGATGGCTGTGGA
GGAGATCAACAATGGATCTGCCTTGCTCCCTGGGCTGCGGCTGGGCTA
TGACCTATTTGACACATGCTCCGAGCCAGTGGTCACCATGAAATCCAG
TCTCATGTTCCTGGCCAAGGTGGGCAGTCAAAGCATTGCTGCCTACTG
CAACTACACACAGTACCAACCCCGTGTGCTGGCTGTCATCGGCCCCCA
CTCATCAGAGCTTGCCCTCATTACAGGCAAGTTCTTCAGCTTCTTCCTC
ATGCCACAGGTCAGCTATAGTGCCAGCATGGATCGGCTAAGTGACCGG
GAAACGTTTCCATCCTTCTTCCGCACAGTGCCCAGTGACCGGGTGCAG
CTGCAGGCAGTTGTGACTCTGTTGCAGAACTTCAGCTGGAACTGGGTG
GCCGCCTTAGGGAGTGATGATGACTATGGCCGGGAAGGTCTGAGCATC
TTTTCTAGTCTGGCCAATGCACGAGGTATCTGCATCGCACATGAGGGC
CTGGTGCCACAACATGACACTAGTGGCCAACAGTTGGGCAAGGTGCTG
GATGTACTACGCCAAGTGAACCAAAGTAAAGTACAAGTGGTGGTGCTG
TTTGCCTCTGCCCGTGCTGTCTACTCCCTTTTTAGTTACAGCATCCATCA
TGGCCTCTCACCCAAGGTATGGGTGGCCAGTGAGTCTTGGCTGACATC
TGACCTGGTCATGACACTTCCCAATATTGCCCGTGTGGGCACTGTGCTT
GGGTTTTTGCAGCGGGGTGCCCTACTGCCTGAATTTTCCCATTATGTGG
AGACTCACCTTGCCCTGGCCGCTGACCCAGCATTCTGTGCCTCACTGAA
TGCGGAGTTGGATCTGGAGGAACATGTGATGGGGCAACGCTGTCCACG
GTGTGACGACATCATGCTGCAGAACCTATCATCTGGGCTGTTGCAGAA
CCTATCAGCTGGGCAATTGCACCACCAAATATTTGCAACCTATGCAGC
TGTGTACAGTGTGGCTCAAGCCCTTCACAACACCCTACAGTGCAATGT
CTCACATTGCCACGTATCAGAACATGTTCTACCCTGGCAGCTCCTGGA
GAACATGTACAATATGAGTTTCCATGCTCGAGACTTGACACTACAGTT

TGATGCTGAAGGGAATGTAGACATGGAATATGACCTGAAGATGTGGGT
GTGGCAGAGCCCTACACCTGTATTACATACTGTGGGCACCTTCAACGG
CACCCTTCAGCTGCAGCAGTCTAAAATGTACTGGCCAGGCAACCAGGT
GCCAGTCTCCCAGTGTTCCCGCCAGTGCAAAGATGGCCAGGTTCGCCG
AGTAAAGGGCTTTCATTCCTGCTGCTATGACTGCGTGGACTGCAAGGC
GGGCAGCTACCGGAAGCATCCAGATGACTTCACCTGTACTCCATGTAA
CCAGGACCAGTGGTCCCCAGAGAAAAGCACAGCCTGCTTACCTCGCAG
GCCCAAGTTTCTGGCTTGGGGGGAGCCAGTTGTGCTGTCACTCCTCCTG
CTGCTTTGCCTGGTGCTGGGTCTAGCACTGGCTGCTCTGGGGCTCTCTG
TCCACCACTGGGACAGCCCTCTTGTCCAGGCCTCAGGTGGCTCACAGT
TCTGCTTTGGCCTGATCTGCCTAGGCCTCT'TCTGCCTCAGTGTCCTTCTG
TTCCCAGGGCGGCCAAGCTCTGCCAGCTGCCTTGCACAACAACCAATG
GCTCACCTCCCTCTCACAGGCTGCCTGAGCACACTCTTCCTGCAAGCAG
CTGAGACCTTTGTGGAGTCTGAGCTGCCACTGAGCTGGGCAAACTGGC
TATGCAGCTACCTTCGGGGACTCTGGGCCTGGCTAGTGGTACTGTTGG
CCACTTTTGTGGAGGCAGCACTATGTGCCTGGTATTTGATCGCTTTCCC
ACCAGAGGTGGTGACAGACTGGTCAGTGCTGCCCACAGAGGTACTGG
AGCACTGCCACGTGCGTTCCTGGGTCAGCCTGGGCTTGGTGCACATCA
CCAATGCAATGTTAGCTTTCCTCTGCTTTCTGGGCACTTTCCTGGTACA
GAGCCAGCCTGGCCGCTACAACCGTGCCCGTGGTCTCACCTTCGCCAT
GCTAGCTTATTTCATCACCTGGGTCTCTTTTGTGCCCCTCCTGGCCAAT
GTGCAGGTGGCCTACCAGCCAGCTGTGCAGATGGGTGCTATCCTAGTC
TGTGCCCTGGGCATCCTGGTCACCTTCCACCTGCCCAAGTGCTATGTGC
TTCTTTGGCTGCCAAAGCTCAACACCCAGGAGTTCTTCCTGGGAAGGA
ATGCCAAGAAAGCAGCAGATGAGAACAGTGGCGGTGGTGAGGCAGCT
CAGGGACACAATGAATGA
The geonomic DNA sequence of the mouse SAC 1 gene (SEQ ID NO: 2) is:
ATCTGAGCCTTAGACACAGCACTGGTGCCAGGCAAACACTCCTGGGCC
TACATGCTTGGG

GCCTCTTCATATTCCAAAAGCTGTCTTTGGGTAAGATGAAGTTCCTCTG
GCAGTGGCATG
AGTGCTGAAGGCTCTTTCCCTGCCCTTCACCTGCTTTCTTGATAGTCTCT
CTGCATACCA
AACAGGCCCTTGTCTCCTGGGAAATGGAAACTATGAAATCAATAGCTG
AGGCTTCTCTAG
GAAAGCCTGCCCTGGTCAGTACAACCTGTTTCACAGCTTCTATAGAAT
AGTTACATCAGC
CTTCTGAAGATGGCCTCTTAGAGCACATGCACCCCCAAGATTCTAAGA
TGTCAATACTAA
CTGACCAAACCATACCTCTCTAGCCAGCCCTGCTGCTCCTGTTGTCTGG
TACCCAGGTGA
CTGAGGACATGACTGGTGGAAGGAAACTAGGCCCCTTTGTCTGTCAGA
TGGCCATACCCA
GCATGGCTGATGCCCAGTGTATAAGACCCTACGCTTTTCCACTGGTCTT
AATGTTAAACC
CTAGGACAGTGTCCTCAGCATAGCTGGTGTGTGTGAATGCAAACTTTG
GGGCATATCTCT
TCCATTAAGCACTGTGATATATGTAGTATTTCCAACAAATAAATTATAC
CTACATGATTG
GGTATAGCATTCTGGGATGGGTCACAGGTGTGTCAGGTGCCTAATTAT
GTGGGGGAAGAA
CATAGAAATATATAGGTGGGGAGGGAGCTAACCCTAGGAATAAGGCT
AAAGCATGTGTCT
CCAGTCCTGAAGACTCAAAGGGCAACGTGAATCATGAGACATGTTCAG
GACTGAAGGAGT
TGCCATGTATCTGTCCTTGATGTATCTTAATCATACATACACTATGAGA
TCTGTGTTACC
TCCATTTTGCAGGTGAGAAAAGAAACACCTGAATGGCCTACCTTAAAG
GGCTAAGTGGGA
AAATAGGTCTGAAGATAACCCAGGCACTGTGTGACAAAGCGGGAAGA
AAACTAGAGATGC

TTTCTTCATGGCAACAACCTAGAGGGTACAACCTAGTGGTTTCTTCTTG
GTACTCCACTG
TATACACCCCATCTGCTTGGGCTGTACATTGTCTGACCATGCTTATAAC
AAAAGTCACAT
ACTACTAGCCAAGACTGAGAACTTAGAGCGACTGGCCAGAAAGTAAA
GATACAACAGTTG
ATATGTGTGCCACACACAGATCCATGTGTACATGTCTATTAATTATGTG
AACGTGCTTTG
TGGACATCCTCACAAAGCAGCAGGGAAATGCAAAGGTCATTTCCATAA
CACCTGCTGGAC
ACCATATGACATTGAGATTACCGGGGTGCCCATTCCAACAAGAGTTAA
TAGCTCCCCCTA
TGTTTGGGTGCCAGAAACCTGATTTGTTAGCAATAGCTCCCTCACATCC
AGATTAAGAGG
GGGATGGCTTAGCTAGGGTTACTATGATGAAACTATGACCAAAGCAAC
TTGTGGGTAAAA
GGGTGTATTTGGCTTACACTTCCATATCACTTCATCAAAGTGAGGACA
GGAACTCAAATA
GAGTAGGAATTTGGTGACAAGAGCTGATGTAGAGGCAATGCAGTGGT
GCCACTTAGTGGC
GCGCTCAGTCTGCTCCCTTTCTTAATAGAATGCAAGACCACCAGCCCAT
GGGTGGCACCA
CAATGGGACCGGGCCCTTCCCCATCGGTCACTAAGAAAATGCCCTACA
GCCAGATCTTAT
GGAGACATTTTCTCAACGGAGGCTCACTCCTTTCAGATAACTCTATATC
AAATTGACATA
AACCAGAACAGAGGAGGAGGCTAAGAAGGAAACTGCCAATTGCATAC
ATGCACACACCTG
GCCCTAGCAGCTGCAGGAAGCTATTTGTTTATGGCCTTTTCTCATTTTC
ATGGACCAGCA
TGAGCACTCTGCAGAGAGAGATGCCTGCATGCCTGCCAAGGCAGGAGT
GCTTACACTGAA

GGTCAACAGGATGGCAGGGGGGCTGCAGAGCTTCCAAGTGTCAGAAC
CCCAGCAGAAGAG
CTGAGACCCTTGCCCGAGGACTCAGGCGGGTTGGGAAGGCCAGGAAA
TTCAGCCAGAGCT
CTTCTTCAGATGGGGTACCATCTGAAGGTTAGACCAGCTAGCCAGCTG
TTGTTGAGGGAC
CACCTCTGCAGCCCCTACCTTTGGAAGATAGAAAGTGTCTCTGTGACA
AGTATGGCCATT
GTGCCCCCTTATTCCACAGTCAACAGAAACCCTGGAATCCTGAACACT
TCTGCAGCTTCT
TTTTTACAGTCTGCCAGGTTGCTCTAGGAATGAAGGGTGCCGAGAGGC
TTGGGCGTAGGC
AGGTGACAAGACCACAGTTAGTGGTCACAGCTGGCTTACTGGATCACT
CTTGGACAGAGT
TTGTTAGATATGGAGTGGAGTATACACAAGGCATCAGGCGGGGGATAT
TGAATGTATCAC
CGGAGCTCCTTGGGGCTTGGCAGCCAAGCACAGCAGTGGTTTTGCTAA
ACAAATCCACGG
TTCCCTCCCCTTGACGCAGTACATCTGTGGCTCCAACCCCACACACCCA
CCCATTGTTAG
TGCTGGAGACTTCTACCTACCATGCCAGCTTTGGCTATCATGGGTCTCA
GCCTGGCTGCT
TTCCTGGAGCTTGGGATGGGGGCCTCTTTGTGTCTGTCACAGCAATTCA
AGGCACAAGGG
GACTACATACTGGGCGGGCTATTTCCCCTGGGCTCAACCGAGGAGGCC
ACTCTCAACCAG
AGAACACAACCCAACAGCATCCCGTGCAACAGGTATGGAGGCTAGTA
GCTGGGGTGGGAG
TGAACCGAAGCTTGGCAGCTTTGGCTCCGTGGTACTACCAATCTGGGA
AGAGGTGGTGAT
CAGTTTCCATGTGGCCTCAGGTTCTCACCCCTTGGTTTGTTCCTGGCCA
TGGCTATGAAG

ATGGCTGTGGAGGAGATCAACAATGGATCTGCCTTGCTCCCTGGGCTG
CGGCTGGGCTAT
GACCTATTTGACACATGCTCCGAGCCAGTGGTCACCATGAAATCCAGT
CTCATGTTCCTG
GCCAAGGTGGGCAGTCAAAGCATTGCTGCCTACTGCAACTACACACAG
TACCAACCCCGT
GTGCTGGCTGTCATCGGCCCCCACTCATCAGAGCTTGCCCTCATTACAG
GCAAGTTCTTC a AGCTTCTTCCTCATGCCACAGGTGAGCCCACTTCCTTTGTGTTCTCAAC
CGATTGCACCC
ATTGAGCTCTCATATCAGAAAGTGCTTCTTGATCACCACAGGTCAGCT
ATAGTGCCAGCA
TGGATCGGCTAAGTGACCGGGAAACGTTTCCATCCTTCTTCCGCACAG
TGCCCAGTGACC
l~ GGGTGCAGCTGCAGGCAGTTGTGACTCTGTTGCAGAACTTCAGCTGGA
ACTGGGTGGCCG
CCTTAGGGAGTGATGATGACTATGGCCGGGAAGGTCTGAGCATCTTTT
CTAGTCTGGCCA
ATGCACGAGGTATCTGCATCGCACATGAGGGCCTGGTGCCACAACATG
ACACTAGTGGCC
AACAGTTGGGCAAGGTGCTGGATGTACTACGCCAAGTGAACCAAAGT
AAAGTACAAGTGG
TGGTGCTGTTTGCCTCTGCCCGTGCTGTCTACTCCCTTTTTAGTTACAGC
ATCCATCATG
GCCTCTCACCCAAGGTATGGGTGGCCAGTGAGTCTTGGCTGACATCTG
ACCTGGTCATGA
CACTTCCCAATATTGCCCGTGTGGGCACTGTGCTTGGGTTTTTGCAGCG
GGGTGCCCTAC
TGCCTGAATTTTCCCATTATGTGGAGACTCACCTTGCCGTGGCCGCTGA
CCCAGCATTCT
GTGCCTCACTGAATGCGGAGTTGGATCTGGAGGAACATGTGATGGGGC
AACGCTGTCCAC

GGTGTGACGACATCATGCTGCAGAACCTATCATCTGGGCTGTTGCAGA
ACCTATCAGCTG
GGCAATTGCACCACCAAATATTTGCAACCTATGCAGCTGTGTACAGTG
TGGCTCAAGCCC
TTCACAACACCCTACAGTGCAATGTCTCACATTGCCACGTATCAGAAC
ATGTTCTACCCT
GGCAGGTAAGGGTAGGGTTTTTTGCTGGGTTTTGCCTGCTCCTGCAGG
AACACTGAACCA
GGCAGAGCCAAATCTTGTTGTGACTGGAGAGGCCTTACCCTGACTCCA
CTCCACAGCTCC
TGGAGAACATGTACAATATGAGTTTCCATGCTCGAGACTTGACACTAC
AGTTTGATGCTG
AAGGGAATGTAGACATGGAATATGACCTGAAGATGTGGGTGTGGCAG
AGCCCTACACCTG
TATTACATACTGTGGGCACCTTCAACGGCACCCTTCAGCTGCAGCAGT
CTAAAATGTACT
GGCCAGGCAACCAGGTAAGGACAAGACAGGCAAAAAGGATGGTGGGT
AGAAGCTTGTCGG
TCTTGGGCCAGTGCTAGCCAAGGGGAGGCCTAACCCAAGGCTCCATGT
ACAGGTGCCAGT
CTCCGAGTGTTCCCGCCAGTGCAAAGATGGCCAGGTTCGCCGAGTAAA
GGGCTTTCATTC
CTGCTGCTATGACTGCGTGGACTGCAAGGCGGGCAGCTACCGGAAGCA
TCCAGGTGAACC
GTCTTCCCTAGACAGTCTGCACAGCCGGGCTAGGGGGCAGAAGCATTC
AAGTCTGGCAAG
CGCCCTCCCGCGGGGCTAATGTGGAGACAGTTACTGTGGGGGCTGGCT
GGGGAGGTCGGT
CTCCCATCAGCAGACCCCACATTACTTTTCTTCCTTCCATCACTACAGA
TGACTTCACCT
GTACTCCATGTAACCAGGACCAGTGGTCCCCAGAGAAAAGCACAGCCT
GCTTACCTCGCA

GGCCCAAGTTTCTGGCTTGGGGGGAGCCAGTTGTGCTGTCACTCCTCCT
GCTGCTTTGCC
TGGTGCTGGGTCTAGCACTGGCTGCTCTGGGGCTCTCTGTCCACCACTG
GGACAGCCCTC
TTGTCCAGGCCTCAGGTGGCTCACAGTTCTGCTTTGGCC.TGATCTGCCT
AGGCCTCTTCT
GCCTCAGTGTCCTTCTGTTCCCAGGGCGGCCAAGCTCTGCCAGCTGCCT
TGCACAACAAC
CAATGGCTCACCTCCCTCTCACAGGCTGCCTGAGCACACTCTTCCTGCA
AGCAGCTGAGA
CCTTTGTGGAGTCTGAGCTGCCACTGAGCTGGGCAAACTGGCTATGCA
GCTACCTTCGGG
GACTCTGGGCCTGGCTAGTGGTACTGTTGGCCACTTTTGTGGAGGCAG
CACTATGTGCCT
GGTATTTGATCGCTTTCCCACCAGAGGTGGTGACAGACTGGTCAGTGC
TGCCCACAGAGG
TACTGGAGCACTGCCACGTGCGTTCCTGGGTCAGCCTGGGCTTGGTGC
ACATCACCAATG
CAATGTTAGCTTTCCTCTGCTTTCTGGGCACTTTCCTGGTACAGAGCCA
GCCTGGCCGCT
ACAACCGTGCCCGTGGTCTCACCTTCGCCATGCTAGCTTATTTCATCAC
CTGGGTCTCTT
TTGTGCCCCTCCTGGCCAATGTGCAGGTGGCCTACCAGCCAGCTGTGC
AGATGGGTGCTA
TCCTAGTCTGTGCCCTGGGCATCCTGGTCACCTTCCACCTGCCCAAGTG
CTATGTGCTTC
TTTGGCTGCCAAAGCTCAACACCCAGGAGTTCTTCCTGGGAAGGAATG
CCAAGAAAGCAG
CAGATGAGAACAGTGGCGGTGGTGAGGCAGCTCAGGGACACAATGAA
TGACCACTGACCC
GTGACCTTCCCTTTAGGGAACCTAGCCCTACCAGAAATCTCCTAAGCC
AACAAGCCCCGA

ATAGTACCTCAGCCTGAGACGTGAGACACTTAACTATAGACTTGGACT
CCACTGACCTTA
GCCTCACAGTGACCCCTTCCCCAAACCCCCAAGGCCTGCAGTGCACAA
GATGGACCCTAT
GAGCCCACCTATCCTTTCAAAGCAAGATTATCCTTGATCCTATTATGCC
CACCTAAGGCC
TGCCCAGGTGACCCACAAA.AGGTTCTTTGGGACTTCATAGCCATACTTT
GAATTCAGAAA
TTCCCCAGGCAGACCATGGGAGACCAGAAGGTACTGCTTGCCTGAACA
TGCCCAGCCCTG
AGCCCTCACTCAGCACCCTGTCCAGGCGTCCCAGGAATAGAAGGCTGG
GCATGTATGTGT
GTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTATGTACGTATG
TATGTATGTAT
CAGGACAGAACAAGAAAGACATCAGGCAGAGGACACTCAGGAGGTAG
GCAACATCCAGCC
TTCTCCATCCCTAGCTGAGCCCTAGCCTGTAGGAGAGAACCAGGTCGC
CGCCAGCACCTT
GGACAGATCACACACAGGGTGCGGGTCAGCACCACGGCCAGCGCCAG
CCACGCGGGACCC
CTGGAATCAGCTTCTAGTACCAAGGACAGAAAAGTTGCCGCAAGGCCC
CTTACTGGCCAG
CACCAGGGACAGAGCCACATGCCTAAGCGGCAAGGGACAAGAGCATC
GTCCATCTGCAGG
CAGGATCAGACCCGGGTCAGTTCTGGACTGGCCCCCACACCTGAATCC
CGGAGCAGCTCA
GCTGGAGAAAAGAGAAACAAGCCACACATCAGTCCCATAAAATTAAA
CGCTTTTTTTAGT
GTTTAAAATAGCATTTACACAGAAGCAGCATTTACACAGAAGCAGCTC
TATGTCAACTAC
CCAGTCACTCAGACTTTGACACAGTGTCTAGTGTAGATGTGTGGGGCC
GCTGTGCCGGGA

TGGCAGTGGCACATGATGATGGGCAGCCACCAGAACAGAAACAGAAC
AGGGCCCAGCTCT
GCAGCTCTTGTGTTCACTGTCACCCACCACTGAGACTGAGACAGTGGC
TAGGTGCCAGGT
CTCTCTCCTGTCTCTCCTACTAGCTACCCTTCACATACCTTCAGTACAA
ACTGTGTTGTC
ATGTGCCAAGTAGCAGGTGGGGAAAGGGGCATGCAAACTGCCCCTTTG
GGTAACTAGCTG
CCACCCTTAGAGCAGGCAGGCTAGCAATAAATAAATAAGTTAGACCCC
ACCTGGGCAGCC
AGAGAGGTTTGAAGGCTCTGTCTAACCCCTCAAAAATCCCACCTTGGC
CTGACAGGTGAG
GCCCATGAACTTAGCGACAGTCAGCCTGTGTCCCTGTGCACAGTTCTGT
GAGGCTTTGGG
GCAAGGGGTACCAAGAGCCCAAGAGAGCCTTTCTTGTTCTAAATGGAG
GTCACTTCCAAA
GAAGGGAACCAGGAGGTGGTCCCTGAGACTTGTGCTGAGGACTTAAA
GTCAGAGATGTCT
CCTTACAAGACTCTATAGATACTTGAGCTGTACCACCATCAGCAGCCC
CAAGAGCAGACA
AAATGTCAAGCCAATATCCTGGTGGTATGGCTGCCCTCAGGCCCTCCT
CTGTAGCCTGCT
CCCTCTGCCCTGGCCCAGAGCCCACAGCTGATCTATCCTGGCTGGCCA
CCACCACGGCCA
GCGCAGAGCTCCTGGCACAGCAGGAGCACAGACTCAGCCACAGGCAG
CGCTGAAGACATT
GGTTGATCATCACATGATGTCCACAAAGAACTCACAGGGGTTTCCCAT
GGCCTTTTGGAA
GGACTGGCGGCTACCTGTAAGTTCTGGAGGGACAGCAGCCAGCTCCCG
GACGGGTGGCCC
TCCAGGTGGCCCACCCACTACTGCATAGGCCTTTGTAAGGGGGTGCAG
TGGGGGGAGCCC

TGGGGCAACAGCTGAAGCCTGACTTCGAGGGCTACTGCCACGGCTAAG
CTGGCTGACAGG
CCGCTCCCACCAGCCGGTGCTACCAGACCCACTTGGTACTGTGTGGTCT
GATTCACTGCC
ACTACCCCCAGCTCCAGTTGCCCGGCGCTCCTCTCGGCCTGGGGTCCG
ATGGCTGCTCCG
TGTGGACCCACTGCTCTTGCTCCCTAGGGGGAGGGAAGGGGACAACAG
AGTCAGCACGAG
GCCTGGCCACTTCCAGGGCCACCAGCTGCTCCCAGACAGTCAGGGCAG
GACCTGGTAAGC
CTGGAGATGGTAGGGGAATGGCAGCCATGCAGATACCAGGAACAGCT
GAGAGGCGAGAAG
CTAGGGGCAGTGGCAGACAGCAGGGACAACAGGGGCCAGCCTGGCAC
CCCACACCTAACC
CCAATGCTTGAACCAAGGGTTAATGTTACAGCTGAGAAACTAAAAACC
AGCGAAGGCCCT
GTGTGCCCAGCATTCCCATTAGCCATCCTGGGTTCACCACCCAAAGAC
CCAACCAGGGTC
CACCCAACCCCAGGACCCTGGTCATCTAATTTGCTTAGCCCCTGTCCTG
AAAGTAGTGGG
AACCTGAAAACACGTGCTGGCTGGGGACATGCTGAGAGGGACACAGG
GGGACCTGGCTTA
CCGGCCCGAGAGTCCACTCTGCTAGTCCTTCAGTCTAAGGCTTGCTCAG
CACAAAGCAAG
GGATAGCACAAGTCACACACCAGTCCAGTGCTCACCAATGGCTAATAG
GACGATTTTGGG
CCAAGCTGAGCCTGGGTACATGCAAGGGCCTGTCCATGGTCAGGATTC
ACTCGATAGCTT
CCCCTTGGGCTTTGCCACCCTCTGGCCCAACCTCTCCTGAGTCTTTCTCT
GGACCTTGTA
GCACAAGTGTGCCCCACTCTGCCTAAGACCTCCACATCAGTCCATCTCC
TCCTGAGGGAC

ACCCACCCTTCAAGATCTTCAATATCCCTGGGATATGCTTTAACACTGA
TATGCTTTAAC
AGTGTTGCTTGATACTCTTATCTGGCACTCTGTTGGGATGCAGGCTCCA
TAACTGATAAA
GCCCATTCTCCCCCTAGCTTGGGGCCTAGAGAGTGCCCCTACCTGCTAT
CAGTGGTTACT
TTCATTCTTGCCATATCATCTCCTGGCCTCTTGCCTCTGCCACCTAGCAC
ACCAGGCTGT
CTTCCTATTCTCTAACGGCTTCTACCCACATCAGCCCCTCCCTGTCCCA
CACACTGACTC
TTGAGATGGAACCCACCGGGACTCAAACACACAGCAGGAGCACAGAG
GGAAGCGTCGGGG
CCAGGCAGAGCGTGGGAGTGGGAGGGAGTGGGAGGAGGGGTGGCAC
GCCTCTCACCTTCA
CTCTGCTGGCTCCCAGCACTGCCGCTGCCGCAGCTGAAGCCAGGGTCC
TGGTAAGCAGGC
GGGAAGCAGGGCGGGGGTCCTGGGTACTGGTAGGGGTAGCCTTGACC
CAAGGGCCAGGGT
ACTGATGGGTGGGGCAGTGGGGCCAGTGTGTCCTGATCTGAGGCTCCA
CTGGAGCCACTG
TTGAGGTTCAGGGATGCGAGGTCTGGCAGGGAGGGAGGGAGGGAGGG
GTAAGTGAAGGCA
AATGAATGAGGCCACAGCAACCCTACCCAACCGCACCCCTACTCACTA
CTGCACAGGTCG
CCAAAGACATAGTAGCACTGCTCAGAAAAGGTGATCTTGTTCACGGTG
TGCCTCAGGAAA
CCGTGCTTCAGCATACTGCTGGCATACTTTCTTGCCTCCCTTCGCTCCTT
GAAGCCCTCC
ACGTGTGTGTACAGCCAGTCCACCACATCCGCCCCTGGCCACAGGTCC
ATCAAAGTCAGG
GTAGCTGAGCCCTGGGAAGCTACGCCAGAATGAGGAACAGACGGGGC
CCTTCCCACACAG

CCAGGGACTCACCAATGACAGCATTGGCAATGGTGATCTTAAGCCACA
TGCGGTCCCGGA
TCTCCAGTCCTGAGTCTGGCAACTGCATGACGCGGACAATGGCACTCA
TGTCACTCTTCA
CAGTCAGCGGTGCCTCCTCAAGCTCTGCAGAGCACACTTCCCTGAGGC
GAGGCTCACAGC
GTGAACCTCCATGGGGTTGAGAGCAGGGGCCAGGGTCAAACCTCTTAT
CTCCCATCCTTG
GGAGATGCCCCTCATCGAAACTTGAGCTAAGACCGGGAGATTCTTCCC
GGTCCCACAGTG
CAAGTCCACGTAGGCAAGGCAGCCCCCCTCCCCTCCCCGGAGAGAACA
AGCTGTTAGCTA
TGTTAGGTAGCAGAA.AAGCAAAGCAGAGGCTGCCATGTCGTCCCAATT
CCCCCCTCCGCA
CAGGCCTGGCAGGACCCTCAATTCATGCAGATGACCAGTATGGCCAGG
CCTGGAGGGATA
TGTACATGTATCTTTGTGTACACATTTGTGAAGGTGTTGGAAGCAAAC
AAAACCTTCATA
TGTAATGGGCCCCTGTAATAGCTCTGATGAGCACCAAAGCTCAAAGCT
AGAACTGACCAT
TGTCCTTCAACCTGAGTTTCCTTGGGTGGGGGGGGGTCCTGTGAGCTGC
CACTTACGTGG
GGCGCCAGGCACTGAGCTGGTTAGTGAGGAAGAGCTGGTGCGTGTGAT
GGCGCTGGAGCA .
GGGACTCGTACCATAGCGGGGCAGGGCACCCGTCAGTGCTGCTGTGTG
GGACAGCCAGGC
AGCCGGGTCGATGGGTCGCACTGGGTCAGCTGCATAGTTTCCAGAGCA
ACGGATTACAGG
TGGTAAGTAGGGGGGCAGCACAGAGGCAGACAAGAAAGACCCCCAGA
CTGAACACAGAAA
CCCCACGCTACCCCACCTTTCCATGGGGTAACTCACCCCTTGGGATGGT
GAAGTAGCTCC

GAGGGGTTGGGTCCCAGCACTTGGCCACTGTGAGACTGATGGGCCTAC
AGAGTTGAGCAG
ACCATGTTGTAAGTGAGGCCCGCACAGCCCCTCCCATCCTGTGCCACT
CCCACCCCCACT
TGGCTCCCACCTCACCCTGTCTGGGACACGATCTCCCGAAGCACCCGT
ACAGCGTCGTCA
TTGCTCATGTTCTCAAAGTTGACATCGTTCACCTACGGGGTTTGTGGGG
TCAGGGGTTGG
TGGTGGGATGTGGGTGCCTCTTGTCCCCACAGTCCCCACATGGCTCCCA
CCTGCAGCAAC
ATGTCGCCCGGCTCAATGCGGCCATCAGCAGCCACGGCCCCGCCCTTC
ATGATGGATCCA
ATGTAGATGCCGCCATCACCCCGGTCGTTGCTCTGGCCCACGATGCTG
ATGCCCAGGAAG
TGGTGCCTCTCTGCAGGAGGGGCCGTGAGCAGGCCCCCAAAGCTCCCG
AGGCTGTACCCA
CCCCCAGCAGGCACCCACAGCCCACAAGGCCTCACCCATGTTGAGAGT
GACGGTGATGAT
GTTCAGGGACATGGTGGAGTCTGTGATGCTGCTGAAGGAGGATGCCTG
CGGAGGGACCCA
GTGAGGGGCTGTGTGGGCACCATTCAGAGCAGACACCCCACCCACCTG
CTGCCTACCCGG
TCTGTCTGCCTCAAGCGCTGCTTCCGACGACGGCATTTGTGCTTCCGAA
CTAGCCGAGAG
GAGGTGCTCTGCTCTGTGGAGCTGCTCAGCCTGAGGCAGGAGTCAGAA
AAGCACAAACAT
GTATAACCAGCTCGGACGCTCAACTACAAATCTCCAGCACGTACTGAC
ATGTGCACACGT
CACCCACCGGCTCGTATTGTCCTCCTCATCTGAGTCAATAAAGCTGCTA
GATTCAAGCTC
ACTGCTCAGTACAGTGGATGCACTGTCTGGAGGTAGTCCCAGGTCCCG
CCGCCGATCCCC

TCTCGGGTGCCCATTGGTCCGGGCAGCTGTGGGGACAGTAGGGTGGGT
ACGACTGTGGGA
CTTCAGTCCTAACAGAATGCGGGTGGCCTGTGCATTTCAAAGTTTATGC
AGTAACTCTGG
GGCCACAGGGGCTAGGAGTACCAGGCTGGGACCTCTACCCAAGGATC
ACTGCTTGGAAGA
ATATGTGGAATACTTCCAGGCTTGGAGTATACCAAAGGGATACCAAG
GG
The polypeptide sequence of mouse SAC1 (SEQ ID NO: 3) is:
MPALAIMGLSLAAFLELGMGASLCLSQQFKAQGDYILGGLFPLGSTEEAT
LNQRTQPNSIPCNRFSPLGLFLAMAMKMAVEEINNGSALLPGLRLGYDLF
DTCSEPVVTMKSSLMFLAKVGSQSIAAYCNYTQYQPRVLAVIGPHSSELA
LITGKFFSFFLMPQVSYSASMDRLSDRETFPSFFRTVPSDRVQLQAVVTLL
QNFSWNWVAALGSDDDYGREGLSIFSSLANARGICIAHEGLVPQHDTSGQ
QLGKVLDVLRQVNQSKVQVVVLFASARAVYSLFSYSIHHGLSPKVWVAS
ESWLTSDLVMTLPNIARVGTVLGFLQRGALLPEFSHYVETHLALAADPAF
CASLNAELDLEEHVMGQRCPRCDDIMLQNLS S GLLQNLSAGQLHHQIFAT
YAAVYSVAQALHNTLQCNVSHCHVSEHVLPWQLLENMYNMSFHARDLT
LQFDAEGNVDMEYDLKMWVWQSPTPVLHTVGTFNGTLQLQQSKMYWP
GNQVPVSQCSRQCKDGQVRRVKGFHSCCYDCVDCKAGSYRKHPDDFTC
TPCNQDQWSPEKSTACLPRRPKFLAWGEPVVLSLLLLLCLVLGLALAALG
LSVHHWDSPLVQASGGSQFCFGLICLGLFCLSVLLFPGRPSSASCLAQQPM
AHLPLTGCLSTLFLQAAETFVESELPLSWANWLCSYLRGLWAWLVVLLA
TFVEAALCAWYLIAFPPEVVTDWSVLPTEVLEHCHVRSWVSLGLVHITNA
MLAFLCFLGTFLVQSQPGRYNRARGLTFAMLAYFITWVSFVPLLANVQV
AYQPAV QMGAILV CALGILV TFHLPKCYVLLWLPKLNTQEFFLGRNAKK
AADENSGGGEAAQGHNE
The cDNA of human SAC 1 (SEQ ID NO: 4) is:
ATGCTGGGCCCTGCTGTCCTGGGCCTCAGCCTCTGGGCTCTCCTGCACC
CTGGGACGGGGGCCCCATTGTGCCTGTCACAGCAACTTAGGATGAAGG

GGGACTACGTGCTGGGGGGGCTGTTCCCCCTGGGCGAGGCCGAGGAG
GCTGGCCTCCGCAGCCGGACACGGCCCAGCAGCCCTGTGTGCACCAGG
TTCTCCTCAAACGGCCTGCTCTGGGCACTGGCCATGAAAATGGCCGTG
GAGGAGATCAACAACAAGTCGGATCTGCTGCCCGGGCTGCGCCTGGGC
TACGACCTCTTTGATACGTGCTCGGAGCCTGTGGTGGCCATGAAGCCC
AGCCTCATGTTCCTGGCCAAGGCAGGCAGCCGCGACATCGCCGCCTAC
TGCAACTACACGCAGTACCAGCCCCGTGTGCTGGCTGTCATCGGGCCC
CACTCGTCAGAGCTCGCCATGGTCACCGGCAAGTTCTTCAGCTTCTTCC
TCATGCCCCAGGTCAGCTACGGTGCTAGCATGGAGCTGCTGAGCGCCC
GGGAGACCTTCCCCTCCTTCTTCCGCACCGTGCCCAGCGACCGTGTGCA
GCTGACGGCCGCCGCGGAGCTGCTGCAGGAGTTCGGCTGGAAGTGGGT
GGCCGCCCTGGGCAGCGACGACGAGTACGGCCGGCAGGGCCTGAGCA
TCTTCTCGGCCCTGGCCTCGGCACGCGGCATCTGCATCGCGCACGAGG
GCCTGGTGCCGCTGCCCCGTGCCGATGACTCGCGGCTGGGGAAGGTGC
AGGACGTCCTGCACCAGGTGAACCAGAGCAGCGTGCAGGTGGTGCTG
CTGTTCGCCTCCGTGCACGCCGCCCACGCCCTCTTCAACTACAGCATCA
GCAGCAGGCTCTCGCCCAAGGTGTGGGTGGCCAGCGAGGCCTGGCTGA
CCTCTGACCTGGTCATGGGGCTGCCCGGCATGGCCCAGATGGGCACGG
TGCTTGGCTTCCTCCAGAGGGGTGCCCAGCTGCACGAGTTCCCCCAGT
ACGTGAAGACGCACCTGGCCCTGGCCACCGACCCGGCCTTCTGCTCTG
CCCTGGGCGAGAGGGAGCAGGGTCTGGAGGAGGACGTGGTGGGCCAG
CGCTGCCCGCAGTGTGACTGCATCACGCTGCAGAACGTGAGCGCAGGG
CTAAATCACCACCAGACGTTCTCTGTCTACGCAGCTGTGTATAGCGTG
GCCCAGGCCCTGCACAACACTCTTCAGTGCAACGCCTCAGGCTGCCCC
GCGCAGGACCCCGTGAAGCCCTGGCAGCTCCTGGAGAACATGTACAAC
CTGACCTTCCACGTGGGCGGGCTGCCGCTGCGGTTCGACAGCAGCGGA
AACGTGGACATGGAGTACGACCTGAAGCTGTGGGTGTGGCAGGGCTC
AGTGCCCAGGCTCCACGACGTGGGCAGGTTCAACGGCAGCCTCAGGAC
AGAGCGCCTGAAGATCCGCTGGCACACGTCTGACAACCAGAAGCCCGT
GTCCCGGTGCTCGCGGCAGTGCCAGGAGGGCCAGGTGCGCCGGGTCA
AGGGGTTCCACTCCTGCTGCTACGACTGTGTGGACTGCGAGGCGGGCA
GCTACCGGCAAAACCCAGACGACATCGCCTGCACCTTTTGTGGCCAGG

ATGAGTGGTCCCCGGAGCGAAGCACACGCTGCTTCCGCCGCAGGTCTC
GGTTCCTGGCATGGGGCGAGCCGGCTGTGCTGCTGCTGCTCCTGCTGCT
GAGCCTGGCGCTGGGCCTTGTGCTGGCTGCTTTGGGGCTGTTCGTTCAC
CATCGGGACAGCCCACTGGTTCAGGCCTCGGGGGGGCCCCTGGCCTGC
TTTGGCCTGGTGTGCCTGGGCCTGGTCTGCCTCAGCGTCCTCCTGTTCC
CTGGCCAGCCCAGCCCTGCCCGATGCCTGGCCCAGCAGCCCTTGTCCC
ACCTCCCGCTCACGGGCTGCCTGAGCACACTCTTCCTGCAGGCGGCCG
AGATCTTCGTGGAGTCAGAACTGCCTCTGAGCTGGGCAGACCGGCTGA
GTGGCTGCCTGCGGGGGCCCTGGGCCTGGCTGGTGGTGCTGCTGGCCA
TGCTGGTGGAGGTCGCACTGTGCACCTGGTACCTGGTGGCCTTCCCGC
CGGAGGTGGTGACGGACTGGCACATGCTGCCCACGGAGGCGCTGGTG
CACTGCCGCACACGCTCCTGGGTCAGCTTCGGCCTAGCGCACGCCACC
AATGCCACGCTGGCCTTTCTCTGCTTCCTGGGCACTTTCCTGGTGCGGA
GCCAGCCGGGCCGCTACAACCGTGCCCGTGGCCTCACCTTTGCCATGC
TGGCCTACTTCATCACCTGGGTCTCCTTTGTGCCCCTCCTGGCCAATGT
GCAGGTGGTCCTCAGGCCCGCCGTGCAGATGGGCGCCCTCCTGCTCTG
TGTCCTGGGCATCCTGGCTGCCTTCCACCTGCCCAGGTGTTACCTGCTC
ATGCGGCAGCCAGGGCTCAACACCCCCGAGTTCTTCCTGGGAGGGGGC
CCTGGGGATGCCCAAGGCCAGAATGACGGGAACACAGGAAATCAGGG
GAAACATGAGTGA
The polypeptide sequence of human SAC1 substantially from the translated region of the human cDNA (SEQ ID NO: 5) is:
MLGPAVLGLSLWALLHPGTGAPLCLSQQLRMKGDYVLGGLFPLGEAEEA
GLRSRTRPSSPVCTRFSSNGLLWALAMKMAVEEINNKSDLLPGLRLGYDL
FDTCSEPWAMKPSLMFLAKAGSRDIAAYCNYTQYQPRVLAVIGPHSSEL
AMVTGKFFSFFLMPQVSYGASMELLSARETFPSFFRTVPSDRVQLTAAAE
LLQEFGWNWVAALGSDDEYGRQGLSIFSALASARGICIAHEGLVPLPRAD
DSRLGKVQDVLHQVNQSSVQVVLLFASVHAAHALFNYSISSRLSPKVWV
ASEAWLTSDLVMGLPGMAQMGTVLGFLQRGAQLHEFPQYVKTHLALAT
DPAFCSALGEREQGLEEDVVGQRCPQCDCITLQNVSAGLNHHQTFSVYAA
VYSVAQALHNTLQCNASGCPAQDPVKPWQLLENMYNLTFHVGGLPLRF

DSSGNVDMEYDLKLWVWQGSVPRLHDVGRFNGSLRTERLKIRWHTSDN
QKPVSRCSRQCQEGQVRRVKGFHSCCYDCVDCEAGSYRQNPDDIACTFC
GQDEWSPERSTRCFRRRSRFLAWGEPAVLLLLLLLSLALGLVLAALGLFV
HHRDSPLVQASGGPLACFGLVCLGLVCLSVLLFPGQPSPARCLAQQPLSHL
PLTGCLSTLFLQAAEIFVESELPLSWADRLSGCLRGPWAWLVVLLAMLVE
VALCT WYLVAFPPEV V TDWHMLPTEALVHCRTRS W V SFGLAHATNATL
AFLCFLGTFLVRSQPGRYNRARGLTFAMLAYFITWVSFVPLLANVQVVLR
PAVQMGALLLCVLGILAAFHLPRCYLLMRQPGLNTPEFFLGGGPGDAQG
QNDGNTGNQGKHE
III. SAC.l Is a G-Protein Coupled Receptor The evidence that SAC is a G-protein coupled receptor (GPCR) comes from its sequence homology to other GPCR and the structure predicted for the amino acid sequence.
GPCRs (also known as 7-transmembrane receptors) bind extracellular ligands and transduce signals into the cell by coupling to intracellular G-proteins.
GPCRs can be subdivided into more than 30 families on the basis of their ligands.
Sac is most closely allied by sequence homology with the Cad-sensing, metabotropic receptors.
Proteins often contain several modules or domains, each with a distinct evolutionary origin and function. When the Sac cDNA sequence is queried against the Conserved Domain Database at NCBI, the following results are obtained:
Sequences producing Score E
significant alignments:

(bits)Value Gnl ~ Pfam ~ pfam01094ANF receptor, Receptor family145 73-36 ligand binding region Gnl ~ Pfam ~ pfam000037tm 3, 7-transmembrane receptor87.0 3e-18 (metabotropic glutamate family) Note the ANF receptor family contains the metabotropic and calcium-sensing families of GCPs.

The closest sequence homology of the mouse SAC gene is to the Cad sensing receptors, all of which are GCPRs. An alignment between a calcium sensing GPCR (BAA09453) is shown in Fig. 5.
As described above, all GPCRs have a characteristic 7-transmembrane domain. Figure 6 is a plot of the transmembrane domains of SAC 1.

r N
p ,G' V ' ~ ~ bA bD p ...'~'', m p O t0., '.~," ~ ',~"'' ~ ,-r-n ~ ~ N _ i~, U C~'/~ "_~ > ~c'~ ~ Pa N V~
Nw ~~+~ ~ ~z N
.t.'~ 'C H ~ N ~ ~b ~~ ,~ N Vi ~ ~
v°' o w o ~ °' +' y '" ,~ +a p N p p ~ GD ~ ~ "C c~~ U
'~ ~ U ° ~~ ~ [~ C% v 0 o O~ .~M.~ ~ ~,'''.~,~ ~v~W
~r,' ~~ N ~ ~ ~ 3 ~ U N
C~ :a ~ ~ M ~j .fl ~ ~ 1n ~~. , ~ V by ~ yr U ~ O V.
a~ a~ o x W $ N b ~ s7 m v~ a~ o ~
p, o.~. ~ N >, N ~ ~ .~-~ ~ ~ ~ b0 y ,-~3,' f cd N U p _~ ~' 'O ~ ~ CA ~ ai ~~ ~ 'L
N ~ 'f" ~ r~ V~ N V O '~' ~ H ~i ~~'' bIl N a 'd ~ bD ~ p., b .~ '~ ,.~~ N ,.~., '"' N N ~ '~ ~1 N tr" ~ N N ~~ U O ~ O
° .o .-b ~ ~o"' .0 3 ~ o.' ~ w ~ E~ ~ ~ O H ~ ~
~1. ci., x ~_ r, C/~ .D u) N x c~ ~ ~ ~ 3 ~ ~ ~a V
c~0,'~ O ~N~yW~y~UO C,' ~"~~tpi~ ~N~ U a ;,~ "O ~ by ~ y y~,~ Pa ~ ~ °~ ~ y ,.°~~ ~ y N ~ p ~ Q
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IV. The Sac Locus and the Gpr98 Sweet Taste Receptor Gene A substantial effort has been devoted to positional cloning of a locus on distal Chr 4 with a major effect on sweetener intake. This locus has been previously described as the Sac (saccharin preference) locus, and it also explains ~8 % of the phenotypic variance in ethanol preferences within the B6 x 129 F2 generation.
Details on positional cloning of the Sac locus are found above.
The effects of SAC1 (Gpr98) on ethanol intake Two lines of evidence point to the involvement of Gpr98 in ethanol intake. First, 129.B6-Sac congenic mice homozygous for a 194-kb donor fragment from the B6 strain consumed more 10% ethanol solution than did congenic mice without the donor fragment (1.50 ~ 0.15 and 1.19 ~ 0.11 mL/day, respectively; p <0.05, one-tailed t-test).
Second, ethanol preference was related to sequence variations of Gpr98.
Analysis of Gpr98 sequences from genealogically remote or unrelated mouse strains indicated the presence of two haplotypes of single nucleotide polymorphisms within the Gpr98 locus. One, 'B6-like' haplotype, was found in mouse strains with elevated sweetener preference and the other, ' 129-like' haplotype, was found in strains relatively indifferent to sweeteners as described above. Preferences for 10% ethanol for the same mouse strains were studied as described in Abstr. of the 23th RSA Meeting (June 2000, Denver, Colorado). We found that strains with the 'B6-like' haplotype had higher preferences for 10% ethanol (20 ~ 4%, n =14, strains C57BL/6J, C57L/J, CAST, FVB/NJ, KK/HIJ, NOD/LtJ, NZB/B1NJ, P/J, RBF/DnJ, RF/J, SEA/GnJ, SJL/J, SPRET/Ei and SWR/J) compared with strains having the ' 129-like' haplotype ( 12 ~ 2%, n =10, p <0.05, one-tailed t-test, strains 129P3/J, AKR/J, BALB/c, BUB/BnJ, C3H/HeJ, CBA/J, DBAl2J, LP/J, PLIJ and RIIIS/J).
V. Preparation of Recombinant or Chemicall~ynthesized Nucleic Acids, Vectors, Transformation, Host-Cells Large amounts of the polynucleotides of the present invention may be produced by replication in a suitable host cell. Natural or synthetic polynucleotide fragments coding for a desired fragment will be incorporated into recombinant polynucleotide constructs, usually DNA constructs, capable of introduction into and replication in a prokaryotic or eukaryotic cell. Usually the polynucleotide constructs will be suitable for replication in a unicellular host, such as yeast or bacteria, but may also be intended for introduction to (with and without integration within the genome) cultured mammalian or plant or other eukaryotic cell lines. The purification of nucleic acids produced by the methods of the present invention is described, e.g., in Ausubel et al., Current Protocols i~
Molecular Biology, Vol. 1-2, John Wiley & Sons, 1992 and Sambrook et al., Molecular CloningA Laboratory Manual, 2nd Ed., Vols. 1-3, Cold Springs Harbor Press, 1989.
The polynucleotides of the present invention may also be produced by chemical synthesis, e.g., by the phosphoramidite method or the triester method, and may be performed on commercial, automated oligonucleotide synthesizers. A
double-stranded fragment may be obtained from the single-stranded product of chemical synthesis either by synthesizing the complementary strand and annealing the strands together under appropriate conditions or by adding the complementary strand using DNA polymerase with an appropriate primer sequence.
Polynucleotide constructs prepared for introduction into a prokaryotic or eukaryotic host may comprise a replication system recognized by the host, including the intended polynucleotide fragment encoding the desired polypeptide, and will preferably also include transcription and translational initiation regulatory sequences operably linked to the polypeptide encoding segment. Expression vectors may include, for example, an origin of replication or autonomously replicating sequence (ARS) and expression control sequences, a promoter, an enhancer and necessary processing information sites, such as ribosome-binding sites, RNA splice sites, polyadenylation sites, transcriptional terminator sequences, and mRNA stabilizing sequences. Secretion signals may also be included where appropriate, whether from a native SAC 1 protein or from other receptors or from secreted polypeptides of the same or related species, which allow the protein to cross andlor lodge in cell membranes, and thus attain its functional topology, or be secreted from the cell. Such vectors may be prepared by means of standard recombinant techniques well-known in the art and discussed, for example, in Sambrook et al., 1989 or Ausubel et al., 1992.
An appropriate promoter and other necessary vector sequences will be selected so as to be functional in the host, and may include, when appropriate, those naturally associated with SAC1 genes. Examples of workable combinations of cell lines and expression vectors are described in Sambrook et al., 1989 or Ausubel et al., 1992. Many useful vectors are known in the art and may be obtained from commercial vendors. Promoters such as the trp, lac and phage promoters, TRNA promoters and glycolytic enzyme promoters may be used in prokaryotic hosts. Useful yeast promoters include promoter regions for metallothionein, 3-phosphoglycerate kinase or other glycolytic enzymes such as enolase or glyceraldehyde-3-phosphate dehydrogenase, enzymes responsible for maltose and galactose utilization, and others. In addition, the construct may be joined to an amplifiable gene so that multiple copies of the gene may be made.
For appropriate enhancer and other expression control sequences, see also Enhancers and Euka~yotic Geue Expression, New York: Cold Spring Harbor Press, 1983.
See also, e.g., US Patent Nos. 5,691,198; 5,735,500; 5,747,469 and 5,436,146.
Expression and cloning vectors will likely contain a selectable marker, a gene encoding a protein necessary for survival or growth of a host cell transformed with the vector. The presence of this gene ensures growth of only those host cells which express the inserts. Typical selection genes encode proteins that (a) confer resistance fo antibiotics or other toxic substances, e.g., ampicillin, neomycin, methotrcxate, etc.; (b) complement auxotrophic deficiencies; or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli. The choice of the proper selectable marker will depend on the host cell, and appropriate markers for different hosts are well-known in the art.
The vectors containing the nucleic acids of interest can be transcribed in vitro, and the resulting RNA introduced into the host cell by well-known methods, e.g., by injection, or the vectors can be introduced directly into host cells by methods well-known in the art, which vary depending on the type of cellular host, including electroporation; transfection employing calcium chloride, rubidium chloride, calcium phosphate, DEAF-dextran, or other substances;
microprojectile bombardment; lipofection; infection (where the vector is an infectious agent, such as a retroviral genome); and other methods. The introduction of the polynucleotides into the host cell by any method known in the S art, including, inter alia, those described above, will be referred to herein as "transformation." The cells into which have been introduced nucleic acids described above are meant to also include the progeny of such cells.
Large quantities of the nucleic acids and polypeptides of the present invention may be prepared by expressing the SAC1 nucleic acids or portions thereof in vectors or other expression vehicles in compatible prokaryotic or eukaryotic host cells. The most commonly used prokaryotic hosts are strains of Eseherichia coli, although other prokaryotes, such as Bacillus subtilis or Pseudomo~as may also be used.
Mammalian or other eukaryotic host cells, such as those of yeast, filamentous fungi, plant, insect, or amphibian or avian species, may also be useful for production of the proteins of the present invention. Propagation of mammalian cells in culture is per se well-known. Examples of commonly used mammalian host cell lines are VERO and HeLa cells, Chinese hamster ovary (CHO) cells, and W138, BHK, and COS cell lines. An example of a commonly used insect cell line is SF9. However, it will be appreciated by the skilled practitioner that other cell lines may be appropriate, e.g., to provide higher expression, desirable glycosylation patterns, or other features.
Clones are selected by using markers depending on the mode of the vector construction. The marker may be on the same or a different DNA molecule, preferably the same DNA molecule. In prokaryotic hosts, the transformant may be selected, e.g., by resistance to ampicillin, tetracycline or other antibiotics.
Production of a particular product based on temperature sensitivity may also serve as an appropriate marker.
VI. Diagnosis or Screening Genetic analysis of obesity and diabetes and alcoholism or alcohol consumption is often complicated by the lack of a simple diagnostic mark. For example, currently there is no single diagnostic marker for the diagnosis of obesity. Sequence variation of the SACl locus may indicate a predisposition to diabetes, obesity, and alcoholism and may provide a diagnostic mark.
In order to detect the presence of a SAC 1 allele predisposing an individual to obesity, diabetes, or alcoholism, a biological sample may be prepared and analyzed for the presence or absence of susceptibility alleles of SAC1.
Results of these tests and interpretive information may be returned to the health care professionals for communication to the tested individual. Such diagnoses may be performed by diagnostic laboratories. In addition, diagnostic kits may be manufactured and available to health care providers or to private individuals for self diagnosis.
A basic format for sequence or expression analysis is finding sequences in DNA or RNA extracted from affected family members which create abnormal SAC1 gene products or abnormal levels of SACl gene product. The diagnostic or screening method may involve amplification or molecular cloning of the relevant SAC1 sequences. For example, PCR based amplification may be used. Once amplified, the resulting nucleic acid can be sequenced or used as a substrate for DNA probes. Primers and probes specific for the SAC 1 gene sequences may be used to identify SAC1 alleles.
The pairs of single-stranded DNA primers can be annealed to sequences within or surrounding the SAC1 gene in order to prime amplifying DNA synthesis of the SAC 1 gene itself. The set of primers may allow synthesis of both intron and exon sequences. Allele-specific primers can also be used. Such primers anneal only to particular SAC1 mutant alleles, and thus will only amplify a product in the presence of the mutant allele as a template.
In order to facilitate subsequent cloning of amplified sequences, primers may have restriction enzyme site sequences appended to their 5' ends. Thus, all nucleotides of the primers are derived from SAC 1 sequences or sequences adjacent to SACl, except for the few nucleotides necessary to form a restriction enzyme site. Such enzymes and sites are well-known in the art. The primers themselves can be synthesized using techniques which are well-known in the art.

Generally, the primers can be made using oligonucleotide synthesizers which are commercially available.
The biological sample to be analyzed, such as blood, may be treated, if desired, to extract the nucleic acids. The sample nucleic acid may be prepared in various ways to facilitate detection of the target sequence; e.g., denaturation, restriction digestion, electrophoresis or dot blotting. The region of interest of the target nucleic acid is usually at least partially single-stranded to form hybrids with the probe. If the sequence is double-stranded, the sequence will probably need to be denatured. The target nucleic acid may be also be fragmented to reduce or eliminate the formation of secondary structures. The fragmentation may be performed using a number of methods, including enzymatic, chemical, thermal cleavage or degradation. For example, fragmentation may be accomplished by heat/Mg2+treatment, endonuclease (e.g., DNAase 1) treatment, restriction enzyme digestion, shearing (e.g., by ultrasound) or NaOH treatment.
Many genotyping and expression monitoring methods have been described previously. In general, target nucleic acid and probe are incubated under conditions which forms hybridization complex between the probe and the target sequence. The region of the probes which is used to bind to the target sequence can be made completely complementary to the targeted region of the SAC 1 locus.
Therefore, high stringency conditions may be desirable in order to prevent false positives. However, conditions of high stringency are typically used if the probes are complementary to regions of the chromosome which are unique in the genome. The stringency of hybridization is determined by a number of factors during hybridization and during the washing procedure, including temperature, ionic strength, base composition, probe length, and concentration of formamide.
Under certain circumstances, the formation of higher order hybrids, such as triplexes, quadraplexes, etc. may be desired to provide the means of detecting target sequences.
Detection, if any, of the resulting hybrid is usually accomplished by the use of labeled probes. Alternatively, the probe may be unlabeled, but may be detectable by specific binding with a ligand which is labeled, either directly or indirectly. Suitable labels, and methods for labeling probes and ligands are known in the art, and include, for example, radioactive labels which may be incorporated by known methods (e.g., nick translation, random priming or kinase reaction), biotin, fluorescent groups, chemiluminescent groups (e.g., dioxetanes, particularly triggered dioxetanes), enzymes, antibodies and the like. Variations of this basic scheme are known in the art, and include those variations that facilitate separation of the hybrids to be detected from extraneous materials and/or that amplify the signal from the labeled moiety.
Two-step label amplification methodologies are known in the art. These assays work on the principle that a small ligand (such as digoxigenin, biotin, or the like) is attached to a nucleic acid probe capable of specifically binding SAC 1.
In one example, the small ligand attached to the nucleic acid probe is specifically recognized by an antibody-enzyme conjugate. In one embodiment of this example, digoxigenin is attached to the nucleic acid probe. Hybridization is detected by an antibody-alkaline phosphatase conjugate which turns over a chemiluminescent substrate. In a second example, the small ligand is recognized by a second ligand-enzyme conjugate that is capable of specifically complexing to the first ligand. A well-known embodiment of this example is the biotin-avidin type of interactions.
It is also contemplated within the scope of this invention that the nucleic acid probe assays of this invention will employ a cocktail of nucleic acid probes capable of detecting SAG 1. Thus, in one example to detect the presence of SAC

in a biological sample, more than one probe complementary to SAG 1 is employed.
Predisposition to diabetes, obesity, or alcoholism can be ascertained by testing any fluid or tissue of a human for sequence variations of the SAC1 gene.
For example, a person who has inherited a germline SAC 1 mutation would be prone to develop obesity, diabetes, or alcoholism. This can be determined by testing DNA from any tissue of the person's body. Most simply, blood can be drawn and DNA extracted from the cells of the blood. In addition, prenatal diagnosis can be accomplished by testing fetal cells, placental cells or amniotic cells for mutations of the SAG 1 gene.

The most definitive test for mutations in a candidate locus is to directly compare genomic SAC1 sequences from obese, diabetic, or alcoholic patients, with those from a control population. Alternatively, one could sequence messenger RNA after amplification, e.g., by PCR, thereby eliminating the necessity of determining the exon structure of the candidate gene.
Sequence variations from diabetic, obese, or alcoholic patients falling outside the coding region of SAC1 can be detected by examining the non-coding regions, such as introns and regulatory sequences near or within the SAC1 gene.
An early indication that mutations in noncoding regions are important may come from Northern blot experiments that reveal messenger RNA molecules of abnormal size or abundance in obese or diabetic patients as compared to control individuals.
Alteration of SAC 1 mRNA expression can be detected by any techniques known in the art (see above). These include Northern blot analysis, PCR
amplification, RNase protection, and gene chip analysis. Diminished mRNA
expression indicates an alteration of the wild-type SAC1 gene.
The diabetic, obese, or alcoholic condition can also be detected on the basis of the alteration of wild-type SACl polypeptide. For example, the presence of a SAC 1 gene variant, which produces a protein having a loss of function, or altered function, may directly correlate to an increased risk of obesity or diabetes.
Such variation can be determined by sequence analysis in accordance with conventional techniques. For example, antibodies (polyclonal or monoclonal) may be used to detect differences in, or the absence of, SAC 1 polypeptides.
Antibodies may immunoprecipitate SAC 1 proteins from solution as well as react with SAC 1 protein on Western or immunoblots of polyacrylamide gels. Antibodies may also detect SAC1 proteins in paraffin or frozen tissue sections, using immunocytochemical techniques. Immunoassay include, for example, enzyme linked immunosorbent assays (ELISA), radioimmunoassays (RIA), immunoradiometric assays (IRMA) and immunoenzymatic assays (IEMA), sandwich assays, etc.

Functional assays, such as protein binding determinations, can be used.
Finding a mutant SAC1 gene product indicates alteration of a wild-type SACl gene.
VII. Drug, Sweetener, and Alcohol Preference Screening This invention is also useful for screening compounds by using the SACl polypeptide or binding fragment thereof in any of a variety of drug, sweetener, and alcohol screening techniques.
The SAC1 polypeptide or fragment employed in such a test may either be free in solution, affixed to a solid support, or borne on a cell surface. One method of drug screening utilizes eukaryotic or prokaryotic host cells which are stably transformed with recombinant polynucleotides expressing the polypeptide or fragment, preferably in competitive binding assays. Such cells, either in viable or fixed form, can be used for standard binding assays. One may measure, for example, for the formation of complexes between a SAC1 polypeptide or fragment and the agent being tested, or examine the degree to which the formation of a complex between a SAC1 polypeptide or fragment and a known ligand is interfered with by the agent being tested.
Thus, the present invention provides methods of screening for drugs and sweeteners comprising contacting such an agent with a SAC1 polypeptide or fragment thereof and assaying (i) for the presence of a complex between the agent and the SAC 1 polypeptide or fragment, or (ii) for the presence of a complex between the SAC 1 polypeptide or fragment and a ligand, by methods well-known in the art. In such competitive binding assays the SACl polypeptide or fragment is typically labeled. Free SAC1 polypeptide or fragment is separated from that present in a protein:protein complex, and the amount of free (i.e., uncomplexed) label is a measure of the binding of the agent being tested to SAC1 or its interference with SACl:ligand binding, respectively.
Other suitable techniques for drug, sweetener, and alcohol screening may provide high throughput screening for compounds having suitable binding affinity to the SAC1 polypeptides. For example, large numbers of different small peptide test compounds are synthesized on a solid substrate, such as plastic pins or some other surface. The peptide test compounds are reacted with SAC 1 polypeptide and washed. Bound SAC1 polypeptide is then detected by methods well-known in the art.
Purified SACl can be coated directly onto plates for use in the aforementioned drug screening techniques. However, non-neutralizing antibodies to the polypeptide can be used to capture antibodies to immobilize the SAC1 polypeptide on the solid phase.
This invention also contemplates the use of competitive drug, sweetener, and alcohol screening assays in which neutralizing antibodies capable of specifically binding the SAC1 polypeptide compete with a test compound for binding to the SAC1 polypeptide or fragments thereof. In this manner, the antibodies can be used to detect the presence of any peptide which shares one or more antigenic determinants of the SAC 1 polypeptide.
A further technique for drug, sweetener, and alcohol screening involves the use of host eukaryotic cell lines or cells which have a nonfunctional SAC

gene. These host cell lines or cells are defective at the SAC 1 polypeptide level.
The host cell lines or cells are grown in the presence of the drug, sweetener, or alcohol compound. The rate of growth of the host cells is measured to determine if the compound is capable of regulating the growth of SAC1 defective cells.
Briefly, a method of screening for a substance which modulates activity of a polypeptide may include contacting one or more test substances with the polypeptide in a suitable reaction medium, testing the activity of the treated polypeptide and comparing that activity with the activity of the polypeptide in comparable reaction medium untreated with the test substance or substances. A
difference in activity between the treated and untreated polypeptides is indicative of a modulating effect of the relevant test substance or substances.
Prior to or as well as being screened for modulation of activity, test substances may be screened for ability to interact with the polypeptide, e.g., in a yeast two-hybrid system. This system may be used as a coarse screen prior to testing a substance for actual ability to modulate activity of the polypeptide.

Alternatively, the screen could be used to screen test substances for binding to a SACl specific binding partner, or to find mimetics of a SAC1 polypeptide.
VIII. Rational Drug Design The goal of rational drug design is to produce structural analogs of biologically active polypeptides of interest or of small molecules with which they interact (e.g., agonists, antagonists, inhibitors) in order to fashion drugs which are, for example, more active or stable forms of the polypeptide, or which, e.g., enhance or interfere with the function of a polypeptide in vivo. In one approach, one first determines the three-dimensional structure of a protein of interest (e.g., SACl polypeptide) or, for example, of the SAC1-receptor or ligand complex, by x-ray crystallography, by computer modeling or most typically, by a combination of approaches. Less often, useful information regarding the structure of a polypeptide may be gained by modeling based on the structure of homologous proteins. An example of rational drug design is the development of HIV
protease inhibitors. In addition, peptides (e.g., SAC1 polypeptide) are analyzed by an alanine scan. In this technique, an amino acid residue is replaced by Ala, and its effect on the peptide's activity is determined. Each of the amino acid residues of the peptide is analyzed in this manner to determine the important regions of the peptide.
It is also possible to isolate a target-specific antibody, selected by a functional assay, and then to solve its crystal structure. In principle, this approach yields a pharmacore upon which subsequent drug design can be based. It is possible to bypass protein crystallography altogether by generating anti-idiotypic antibodies (anti-ids) to a functional, pharmacologically active antibody. As a mirror image of a mirror image, the binding site of the anti-ids would be expected to be an analog of the original receptor. The anti-id could then be used to identify and isolate peptides from banks of chemically or biologically produced banks of peptides. Selected peptides would then act as the pharmacore.
Thus, one may design drugs which have, e.g., improved SACl polypeptide activity or stability or which act as inhibitors, agonists, antagonists, etc.
of SAC1 polypeptide activity. By virtue of the availability of cloned SAC1 sequences, sufficient amounts of the SAC1 polypeptide may be made available to perform such analytical studies as x-ray crystallography. In addition, the knowledge of the SAC1 protein sequence provided herein will guide those employing computer modeling techniques in place of, or in addition to x-ray crystallography.
Following identification of a substance which modulates or affects polypeptide activity, the substance may be investigated further. Furthermore, it may be manufactured andlor used in preparation, i.e., manufacture or formulation, or a composition such as a medicament, pharmaceutical composition or drug.
These may be administered to individuals.
Thus, the present invention extends in various aspects not only to a substance identified using a nucleic acid molecule as a modulator of polypeptide activity, in accordance with what is disclosed herein, but also a pharmaceutical composition, medicament, drug or other composition comprising such a substance, a method comprising administration of such a composition comprising such a substance, a method comprising administration of such a composition to a patient, e.g., for treatment of diabetes, obesity or alcohol consumption, use of such a substance in the manufacture of a composition for administration, e.g., for treatment of diabetes or alcohol consumption, and a method of making a pharmaceutical composition comprising admixing such a substance with a pharmaceutically acceptable excipient, vehicle or carrier, and optionally other ingredients.
A substance identified as a modulator of polypeptide function may be peptide or non-peptide in nature. Non-peptide "small molecules" are often preferred for many in vivo pharmaceutical uses. Accordingly, a mimetic or mimic of the substance (particularly if a peptide) may be designed for pharmaceutical use.
The designing of mimetics to a known pharmaceutically active compound is a known approach to the development of pharmaceuticals based on a "lead"
compound. This might be desirable where the active compound is difficult or expensive to synthesize or where it is unsuitable for a particular method of administration, e.g., pure peptides are unsuitable active agents for oral compositions as they tend to be quickly degraded by proteases in the alimentary canal. Mimetic design, synthesis and testing is generally used to avoid randomly screening large numbers of molecules for a target property.
There are several steps commonly taken in the design of a mimetic from a compound having a given target property. First, the particular parts of the compound that are critical and/or important in determining the target property are determined. In the case of a peptide, this can be done by systematically varying the amino acid residues in the peptide, e.g., by substituting each residue in turn.
Alanine scans of peptide are commonly used to refine such peptide motifs.
These parts or residues constituting the active region of the compound are known as its pharmacophore. .
Once the pharmacophore has been found, its structure is modeled according to its physical properties, e.g., stereochemistry, bonding, size and/or charge, using data from a range of sources, e.g., spectroscopic techniques, x-ray diffraction data and NMR. Computational analysis, similarity mapping (which models the charge and/or volume of a pharmacophore, rather than the bonding between atoms) and other techniques can be used in this modeling process.
In a variant of this approach, the three-dimensional structure of the ligand and its binding partner are modeled. This can be especially used where the ligand and/or binding partner change conformation on binding, allowing the model to take account of this in the design of the mimetic.
A template molecule is then selected onto which chemical groups which mimic the pharmacophore can be grafted. The template molecule and the chemical groups grafted onto it can conveniently be selected so that the mimetic is easy to synthesize, is likely to be pharmacologically acceptable, and does not degrade in vivo, while retaining the biological activity of the lead compound.
Alternatively, where the mimetic is peptide-based, further stability can be achieved by cyclizing the peptide, increasing its rigidity. The mimetic(s) found by this approach can then be screened to see whether they have the target property, or to what extent they exhibit it. Further optimization or modification can then be carried out to arrive at one or more final mimetics for in vivo or clinical testing.

IX. Gene Therapy According to the present invention, a method is also provided of supplying wild-type SAC 1 function to a cell which carries mutant SAC 1 alleles. The wild-type SACl gene or a part of the gene may be introduced into the cell in a vector such that the gene remains extrachromosomal. In such a situation, the gene will be expressed by the cell from the extra chromosomal location. More preferred is the situation where the wild-type SAC1 gene or a part thereof is introduced into the mutant cell in such a way that it recombines with the endogenous mutant SAC1 gene present in the cell. Such recombination requires a double recombination event which results in the correction of the SAC1 gene mutation. Vectors for introduction of genes both for recombination and for extrachromosomal maintenance are known in the art, and any suitable vector may be used. Methods for introducing DNA into cells such as electroporation, calcium phosphate coprecipitation and viral transduction are known in the art, and the choice of method is within the competence of skilled practitioners.
As generally discussed above, the SAC 1 gene or fragment, where applicable, may be employed in gene therapy methods in order to increase the amount of the expression products of such genes in diabetic or obese cells.
Such gene therapy is particularly appropriate, in which the level of SAC 1 polypeptide is absent or compared to normal cells. It may also be useful to increase the level of expression of a given SAC 1 gene even in those situations in which the mutant gene is expressed at a "normal" level, but the gene product is not fully functional.
Gene therapy would be carried out according to generally accepted methods, for example, as described by Therapy for Genetic Diseases, T. Friedman, ed. Oxford University Press, 1991. Cells from a patient would be first analyzed by the diagnostic methods described above, to ascertain the production of SAC1 polypeptide in these cells. A virus or plasmid vector, containing a copy of the SAC 1 gene linked to expression control elements and capable of replicating inside the sample cells, is prepared. Suitable vectors are known, such as disclosed in PCT publications WO 93/07282 and United States PatentNos. 5,252,479, 5,691,198, 5,747,469, 5,436,146 and 5,753,500. The vector is then injected into the patient.
Gene transfer systems known in the art may be useful in the practice of the gene therapy methods of the present invention. These include viral and nonviral transfer methods. A number of viruses have been used as gene transfer vectors, including papovaviruses, e.g., SV40, adenovirus, vaccinia virus, adeno-associated virus, herpes viruses including HSV and EBV; lentiviruses, Sindbis and Semliki Forest virus, and retroviruses of avian, marine, and human origin. Most human gene therapy protocols have been based on disabled marine retroviruses.
Nonviral gene transfer methods known in the art include chemical techniques such as calcium phosphate coprecipitation; mechanical techniques, for example microinjection; membrane fusion-mediated transfer via liposomes; and direct DNA uptake and receptor-mediated DNA transfer. Viral-mediated gene transfer can be combined with direct in vivo gene transfer using liposome delivery, allowing one to direct the viral vectors to the affected cells and not into the surrounding nondividing cells. Alternatively, the retroviral vector producer cell line can be injected into affected cells. Injection of producer cells would then provide a continuous source of vector particles.
In an approach which combines biological and physical gene transfer methods, plasmid DNA of any size is combined with a polylysine-conjugated antibody specific to the adenovirus hexon protein, and the resulting complex is bound to an adenovirus vector. The trimolecular complex is then used to infect cells. The adenovirus vector permits efficient binding, internalization, and degradation of the endosome before the coupled DNA is damaged. For other techniques for the delivery of adenovirus based vectors see United States Patent Nos. 5,691,198; 5,747,469; 5,436,146 and 5,753,500.
Liposome/DNA complexes have been shown to be capable of mediating direct in vivo gene transfer. While in standard liposome preparations the gene transfer process is nonspecific, localized in vivo uptake and expression may be accomplished following direct iii situ administration.
Expression vectors in the context of gene therapy are meant to include those constructs containing sequences sufficient to express a polynucleotide that has been cloned therein. In viral expression vectors, the construct contains viral sequences sufficient to support packaging of the construct. If the polynucleotide encodes SACl, expression will produce SACl. If the polynucleotide encodes an antisense polynucleotide or a ribozyme, expression will produce the antisense polynucleotide or ribozyme. Thus in this context, expression does not require that a protein product be synthesized. In addition to the polynucleotide cloned into the expression vector, the vector also contains a promoter functional in eukaryotic cells. The cloned polynucleotide sequence is under control of this promoter.
Suitable eukaryotic promoters include those described above. The expression vector may also include sequences, such as selectable markers and other sequences described herein.
Receptor-mediated gene transfer, for example, may be accomplished by the conjugation of DNA (usually in the form of covalently closed supercoiled plasmid) to a protein ligand via polylysine. Ligands are chosen on the basis of the presence of the corresponding ligand receptors on the cell surface of the target cell/tissue type. One appropriate receptor/ligand pair may include the estrogen receptor and its ligand, estrogen (and estrogen analogues). These ligand-DNA
conjugates can be injected directly into the blood if desired and are directed to the target tissue where receptor binding and internalization of the DNA-protein complex occurs. To overcome the problem of intracellular destruction of DNA, coinfection with adenovirus can be included to disrupt endosome function.
X. Peptide Therapy Peptides which have SAC 1 activity can be supplied to cells which carry mutant or missing SAC1 alleles. Protein can be produced by expression of the cDNA sequence in bacteria, for example, using known expression vectors.
Alternatively, SAC1 polypeptide can be extracted from SAC1-producing mammalian cells. In addition, the techniques of synthetic chemistry can be employed to synthesize SAC 1 protein. Any of such techniques can provide the preparation of the present invention which comprises the SAC 1 protein.

Preparation is substantially free of other human proteins. This is most readily accomplished by synthesis in a microorganism or in vitro.
Active SAC1 molecules can be introduced into cells by microinjection or by use of liposomes, for example. Alternatively, some active molecules may be taken up by cells, actively or by diffusion. Extra-cellular application of the gene product may be sufficient. Molecules with SAC 1 activity (for example, peptides, drugs or organic compounds) may also be used to effect such a reversal.
Modified polypeptides having substantially similar function are also used for peptide therapy.
XI. Transformed Hosts Similarly, cells and animals which carry a mutant SAC1 allele can be used as model systems to study and test for substances which have potential as therapeutic agents. These may be isolated from individuals with SAC1 mutations, either somatic or germline. Alternatively, the cell line can be engineered to carry the mutation in the SAC1 allele.
Animals for testing therapeutic agents can be selected after mutagenesis of whole animals or after treatment of germline cells or zygotes. Such treatments include insertion of mutant SAC1 alleles, usually from a second animal species, as well as insertion of disrupted homologous genes. Alternatively, the endogenous SAC1 gene of the animals may be disrupted by insertion or deletion mutation or other genetic alterations using conventional techniques to produce knockout or transplacement animals. A transplacement is similar to a knockout because the endogenous gene is replaced, but in the case of a transplacement the replacement is by another version of the same gene. After test substances have been administered to the animals, the phenotype must be assessed. If the test substance prevents or suppresses the disease, then the test substance is a candidate therapeutic agent for the treatment of disease. These animal models provide an extremely important testing vehicle for potential therapeutic products.
In one embodiment of the invention, transgenic animals are produced which contain a functional transgene encoding a functional SACl polypeptide or variants thereof. Transgenic animals expressing SAC1 transgenes, recombinant cell lines derived from such animals and transgenic embryos may be useful in methods for screening for and identifying agents that induce or repress function of SACl . Transgenic animals of the present invention also can be used as models for studying indications such as diabetes.
In one embodiment of the invention, a SAC1 transgene is introduced into a non-human host to produce a transgenic animal expressing a human or marine SAC 1 gene. The transgenic animal is produced by the integration of the transgene into the genome in a manner that permits the expression of the transgene.
Methods for producing transgenic animals are generally described in US Patent No. 4,873,191 and in Manipulating the Mouse Embryo; A Laboratory Manual, 2nd edition (eds., Hogan, Beddington, Costantimi and Long, New York: Cold Spring Harbor Laboratory Press, 1994).
Tt may be desirable to replace the endogenous SAC1 by homologous recombination between the transgene and the endogenous gene; or the endogenous gene may be eliminated by deletion as in the preparation of "knock-out"
animals.
Typically, a SAC 1 gene flanked by genomic sequences is transferred by microinjection into a fertilized egg. The microinjected eggs are implanted into a host female, and the progeny are screened for the expression of the transgene.
Transgenic animals may be produced from the fertilized eggs from a number of animals including, but not limited to reptiles, amphibians, birds, mammals, and fish. Within a particularly preferred embodiment, transgenic mice are generated which express a mutant form of the polypeptide.
As noted above, transgenic animals and cell lines derived from such animals may find use in certain testing experiments. In this regard, transgenic animals and cell lines capable of expressing wild-type or mutant SAC 1 may be exposed to test substances. These test substances can be screened for the ability to reduce overexpression of wild-type SAC 1 or impair the expression or function of mutant SAC 1.

XII. Pharmaceutical Compositions and Routes of Administration The SAC1 polypeptides, antibodies, peptides and nucleic acids of the present invention can be formulated in pharmaceutical compositions, which are prepared according to conventional pharmaceutical compounding techniques. See, for example, Remingtoh's Pharmaceutic. Sciehces, 18th Ed. (Euston, PA: Mack Publishing Co., 1990). The composition may contain the active agent or pharmaceutically acceptable salts of the active agent. These compositions may comprise, in addition to one of the active substances, a pharmaceutically acceptable excipient, carrier, buffer, stabilizer or other materials well-known in the art. Such materials should be nontoxic and should not interfere with the efficacy of the active ingredient. The carrier may take a wide variety of forms depending on the form of preparation desired for administration, e.g., intravenous, oral, intrathecal, epineural or parenteral.
For oral administration, the compounds can be formulated into solid or liquid preparations such as capsules, pills, tablets, lozenges, melts, powders, suspensions or emulsions. In preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, suspending agents, and the like in the case of oral liquid preparations (such as, for example, suspensions, elixirs and solutions); or carriers such as starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like in the case of oral solid preparations (such as, for example, powders, capsules and tablets). Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously employed. If desired, tablets may be sugar-coated or enteric-coated by standard techniques. The active agent can be encapsulated to make it stable to passage through the gastrointestinal tract while at the same time allowing for passage across the blood brain barrier. See for example, 8.
For parenteral administration, the compound may be dissolved in a pharmaceutical carrier and administered as either a solution or a suspension.

Illustrative of suitable carriers are water, saline, dextrose solutions, fructose solutions, ethanol, or oils of animal, vegetative or synthetic origin. The carrier may also contain other ingredients, for example, preservatives, suspending agents, solubilizing agents, buffers and the like. When the compounds are being administered intrathecally, they may also be dissolved in cerebrospinal fluid.
The active agent is preferably administered in a therapeutically effective amount. The actual amount administered, and the rate and time-course of administration, will depend on the nature and severity of the condition being treated. Prescription of treatment, e.g., decisions on dosage, timing, etc., is within the responsibility of general practitioners or specialists, and Typically takes account of the disorder to be treated, the condition of the individual patient, the site of delivery, the method of administration and other factors known to practitioners. Examples of techniques and protocols can be found in Remington's Pharmaceutical Sciences.
Alternatively, targeting therapies may be used to deliver the active agent more specifically to certain types of cell, by the use of targeting systems such as antibodies or cell specific ligands. Targeting may be desirable for a variety of reasons, e.g., if the agent is unacceptably toxic, or if it would otherwise require too high a dosage, or if it would not otherwise be able to enter the target cells.
Instead of administering these agents directly, they could be produced in the target cell, e.g., in a viral vector such as described above or in a cell based delivery system such as described in United States Patent No. 5,550,050 and PCT
publications WO 92/19195, WO 94/25503, WO 95/01203, WO 95/05452, WO 96!02286, WO 96/02646, WO 96/40871, WO 96/40959 and WO 97!12635, designed for implantation in a patient. The vector could be targeted to the specific cells to be treated, or it could contain regulatory elements which are more tissue specific to the target cells. The cell based delivery system is designed to be implanted in a patient's body at the desired target site and contains a coding sequence for the active agent. Alternatively, the agent could be administered in a precursor form for conversion to the active form by an activating agent produced in, or targeted to, the cells to be treated. See for example, EP 425,731A and WO 90/07936.

EXAMPLES
The following examples further illustrate the present invention. These examples are intended merely to be illustrative of the present invention and are not to be construed as being limiting.

Animal care and maintenance. All animal protocols used in these studies were approved by the Monell Institutional Animal Care and Use Committee. Mice were housed in individual cages in a temperature- controlled room at 23°C on a 12-hour light:l2-hour dark cycle. The animals had free access to deionized water and Teklad Rodent Diet 8604 (Harlan Teklad, Madison, WI).

Breeding of F2 and partially congenic mice. C57BL6/ByJ (B6) and 129P3/J (formerly named 129/J; abbreviated here as 129) mice were purchased from The Jackson Laboratory. The B6 and 129 mice were outcrossed to produce the first filial generation of hybrids (Fl), and these were intercrossed to produce the second hybrid generation (F2, n = 629).
To create the partially congenic lines, the F2 mice were genotyped with several markers on the distal part of chromosome 4, and a few F2 mice with recombinations in this region were used as founders of strains partially congenic with the 129 strain. These F2 founders were backcrossed to the 129 strain to produce the N2 generation. Mice from this and subsequent backcross generations were phenotyped using 96-hour two-bottle tests with saccharin solutions, and genotyped using markers on distal chromosome 4 and on other autosomes. Mice with high saccharin intake (with a fragment of distal chromosome 4 from the B6 strain and homozygous for 129 alleles of markers on other chromosomes) were selected for subsequent backcrossing. This marker-assisted selection resulted in a segregating 129.B6-Sac partially congenic strain. Three strains were created, with different overlapping fragments containing the SACl gene.

Testing of sweet preference in the F2 and partially congenic mice.
Consumption of 120 mM sucrose and 17 mM saccharin (Sigma Chemical Company, St. Louis, MO) was measured in individually caged mice using 96-hour S two-bottle tests, with water as the second choice. The positions of the tubes were switched every 24 hours. Fluid intakes were expressed per 30 g of body weight (the approximate weight of an adult mouse) per day, or as a preference score (ratio of average daily solution intake to total fluid intake, in percent).

Genotyping of F2 mice and linkage analysis. Genomic DNA was purified from mouse tails by NaOH/Tris (Beier, personal communication;
Truett G.E. et al., Preparation of PCR-quality mouse genomic DNA with hot sodium hydroxide and tris (HotSHOT) [In Process Citation]. Biotechniques, 2000;29:52, S4), or the phenol/chloroforzn method. All F2 mice were genotyped 1 S with all available polymorphic microsatellite markers (Research Genetics, Huntsville, AL) known to map near the SAC 1 region with a protocol modified slightly from that described by Dietrich W. et al., A genetic map of the mouse suitable for typing intraspecific crosses. Genetics, 1991;131;423-447. The maxkers tested are as follows: D4Mit190, D4Mit42, D4Mit2S4, and D4Mit2S6. Analysis of this framework map using MAPMAKER/QTL 1. I (Lender E. et aI.
MAPMAKER: An interactive complex package for constructing primary linkage maps of experimental and natural populations. Genomics, 1987;1:174-181), indicated that Sac mapped distal to D4Mit2S6, and therefore all available STS
and EST were tested by SSCP (Orita M., Iwahana H., Kanazawa H., Hayashi K., and 2S Sekiya T. Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphins. Proceedings of the Natio~zal Academy of Sciences of the USA, 1989:86) or direct sequencing, for polymorphisms between the B6 and 129 strains. Polymorphisms between strains were found for the following maxkers: D18346, AA410003 (K00231), V2r2, and D4Erdt296E, and the linkage analysis conducted again using these polymorphic maiCers.

Genotyping of partially congenic mice. Three partially congenic strains of mice were genotyped with all available markers, and those markers with two 129 alleles were excluded from the SAC 1 nonrecombinant interval.

Radiation hybrid mapping. To generate additional markers to narrow the Sac nonrecombinant interval, several markers were tested using the T31 RH
genome map. Primers from several sequences suggested through survey of the public databases were constructed and DNA from the T31 panel. Results were scored using software at the Jackson Laboratory.

Construction of BAC contig and marker development. To construct a physical map of the SAC1 region, the RPCI-23 BAC library was screened with markers within and near the SAC1 nonrecombinant interval: each marker was tested by whole cell PCR to confirm its presence. Only those markers positive by both hybridization and PCR are shown. Primers for the BAC ends were constructed from sequence obtained through TIGR (www.ti~r.org) or by direct sequencing, when necessary. Each positive clone was tested for the presence of each BAC end (if the BAC end contained unique sequence), and the contig oriented using SEGMAP, Version 3.48 (Green E.D. and Green P. Sequence-tagged site (STS) content mapping of human chromosomes: theoretical considerations and early experiences. PCR Methods Appl., 1991;1:77-90). BAC
end sequences was amplified in B6 and 129 strains, and analyzed by SSCP or direct sequencing. Those BAC ends polymorphic between 129 and B6 were tested in the recombinant F2 and partially congenic mice, to further narrow the SAC 1 nonrecombinant interval.

Amplification of SACl and polymorphism detection. After the SAC1 nonrecombinant interval was narrowed to less than 350 kb, a 246 kb BAC was chosen for sequencing which spanned most of the region (RPCI-23-118E21).
Within this BAC, there was a gene with homology to other taste receptors.
Using 11.8 kb of sequence and the program GENSCAN (Barge C.B. and Karlin S.
Finding the genes in genomic DNA. Current Opinion Structural Biology, 1998;8:346-354), a 858 amino acid protein, with 6 exons, was identified.
Primers were constructed that amplified this gene, and an additional 2600 nt upstream and 5200 nt downstream were also amplified (primer sequence available upon request). These PCR products were sequenced using genomic DNA from B6 and 129 mouse strains, as well as other strains with either higher (SWR/J, C57L/J, IS, ST/bJ, SEA/GnJ) or lower (DBA/2J, AKR/J, BALB/cByJ) saccharin preference (Lush LE., The genetics of tasting in mice. VI. Saccharin, acesulfame, dulcin and sucrose. Genet Res., 1989;53:95-99; Lush I. The genetics of bitterness, sweetness, and saltiness in strains of mice. in Genetics of perception and communication, Vol. 3, eds. Wysocki C. and Kare M., New York: Marcel Dekker, 1991:227-235;
Lush LE. and Holland G. The genetics of tasting in mice. V. Glycine and cycloheximide. Genet Res., 1988;52:207-12). Sequences were aligned with Sequencer (Gene Codes, Ann Arbor, MI) and the single nucleotide polymorphisms, insertions and deletions identified.

Preparation of tongue cDNA and expression studies. Total RNA was extracted from anterior mouse tongue from the 129 and B6 strains (TRIZOL
Reagent; GIBCOBRL). Total RNA (200 ng) was reverse transcribed using the Life Technologies Superscript Kit. Following the reverse transcription, the samples were amplified using Advantage cDNA PCR Kit (Clontech, Palo Alto, CA). Primers were constructed to span exon 2 and 3, so that the genomic and cDNA product size would differ (Primer set 3A;
Left-5'TGCATTGGCCAGACTAGAAA3';

Right-SCGGCTGGGCTATGACCTAT'). The expected product size for primer 3A is 418 by for cDNA and 497 by for genomic DNA. Single bands of these sizes were excised from the gel, purified and sequenced, confirming the intron-exon boundary and expression of mRNA of this gene in mouse tongue. Primers were then designed to cover the whole cDNA, and, the sequences obtained and aligned, to confirm intronlexon boundaries.

Human gene expression. The human ortholog of the SAC1 gene was examined for mRNA expression in human tongue. Total RNA from human taste papillae was obtained through biopsy, a procedure approved by the Committee on Studies Involving Human Beings at the University of Pennsylvania. The RNA was extracted as described above, reverse transcribed, and amplified, with human specific primers. Two bands were obtained of the expected size for genomic and cDNA. Sequencing of these bands co~rmed the SAC 1 gene is expressed in human taste papillae.

Tissue Expression of SACl. Oligonucleotide primers specific for different parts of the SAC1 gene were used to assay different tissues for SAC1 expression as shown in Table 2. Tissue specific cDNA pools were purchased from OriGene Technologies Ltd. Primer pair 3A, amplifies parts of exons 2 and 3, with a small intron to differentiate between PCR product representing genomic DNA
versus cDNA. Primer pair 6A amplifies parts of exons 4 and 5. This part of the protein encodes the 7TM domain, and may cross react with other GPCRs expressed in different tissues.

Table 2. Expression pattern of SAC1 Tissue 3A 6A

Brain - -Heart - -Kidney + +

Spleen + +

Thymus + +

Liver - +

Stomach - +

Sm Intestine - +

Muscle - +

Lung - +

Testis + +

Skin - -Adrenal + -Pancreas + +

Uterus - -Prostrate + +

Embryo-8.5 - -9.5 - -12.5 - -19 + -/+

Breast-virgin - +

Pregnant - +

Lactating + +

Involuting - -Primers for the SACl Locus (Seq. ID Nos.: 6-651) are:
Marker Forward Reverse Size, SEQ.
by ID

NO.

28.MMHAP7B4.CACTAGAGCTGCC CCCTCAGCACCA 162 6-7 seq ACCTTCC CTTTTTGT

28.MMHAP7B4.ACAAAAAGTGGTG CAGGAGACCCA 163 8-9 seq CTGAGGG AAGGATCAA

GAGGCAC ATAGGTGG

GCG ACTTCTGC

CAGCC TGCG

TGGAG GAGACAC

Agrin4 TGTGACTTCCTCTT TGAGCCACTCCA 156 18-19 CCCCAC GATGTCAG

Agrin4 GTGTGTCAGCATC CCAACGTGCAGT 290 20-21 ACTGCCT CAAGAAAA

Agrin4 CGAGAGACAAAG TTATGAAGGCCC 263 22-23 TGGTGCTG TCACCAAC

Agrin4 CCAGCTCCTAGAA GCAGTCTGCCGA 298 24-25 TTGCCTG AACAAGTC

Agrin4 ATAGAGGAATGG TACCAGGAGGG 299 26-27 GTGCGATG GTCAGTCAG

Agrin4 TACAAGCGAGCTG CCAATCAGCTCG 271 28-29 ACCAATG AGTTAGCC

AgrinA TGCCATTGTGGAT GAGTCCGAGGTC 575 30-31 GTTCACT GGTCAATA

Marker Forward Reverse Size, SEQ.
by ID

NO.

AgrinB GCTGGCTTCTGTA TATGAGGGTCAA 577 32-33 GGTCAGG GGGTCAGG

AgrinC CGCTTTGGTGAGA CATGTGGAGTTG 573 34-35 ACTAGCC TGGGAGTG

AgrinD AATGGGCAGAAG TATCAGGGTCTG 507 36-37 ACAGATGG TGAAGCCC

AgrinE ATACAGGACCCTT CAGTGTTTCTAG 587 38-39 TACCCCG GTCCCCCA

Agrin GCCTCTGTCTGCC ATAATGTTACCT 594 40-41 ATCTCTC GCAGGCGG

TGTCCTC ACACTTTTA

AAGGTGA AGTTGAGA

TCAGGGC AGTTGAGA

AAGGTGA CCTCGGTG

TGCAGACCC ATAGTGCC

Cdc212 GGCTTCAGCCTCA AAAACAACCAA 101 52-53 AGTTCTG GTTGCCCTG

Cdc212 GGCACTGAAATGA AACAATTCAAGC 265 54-55 CCTGGAT AACCTCGG

Cdc212 CTGTTCCTTCCCA TTCAGTCACGCA 225 56-57 GACTCCA AACCTGAG

Cot GCCCAGGACTTTG GGTAACCTGCAG 284 58-59 TCACTGT CTCCACTC

Marker Forward Reverse Size, SEQ.
by ID

NO.

Cot GGGACATGCTCTT GAACAAAGCCG 277 60-61 GGTTCAT GGTGATTTA

Cot GCCCTCAGTTCTC GGCAGAGAAGA 110 62-63 CTAGCCT CTGGTGGAG

Cot CCCAGACTTAGCG AGCAGAGACCTT 277 64-65 TCTCAGG TGGACTCG

Cot GAAGGCTGAGTGA TTGCACGAGGAG 276 66-67 GTCCCAG AAGGTTTT

Cot GATGCCAACGAGA AGAAGCCAAAA 247 68-69 CCTGAAT CCCTCACCT

Cot AAAAAGCCCTGCA ATTCAGGTCTCG 107 70-71 AGAACTT TTGGCATC

GAAACTA GAGAGCCC

CCTGGCT AGTGTCAA

D4Bir1 GCGATGTTGTTG CAGTGTCTTTCC 76-77 CG ACATTT

D4Ertd296e AGGCATATTGTAT CCGGATGACTCT 201 78-79 AATAAATTTGTA ACTTGAC

GT

D4Hrb1 GCTGTTTATGGGG AATTTCTGAAGC 194 80-81 TCGAGAA AGGGGGAT

D4Hrb1 TCCCCCTGCTTCA AGGGGGATGATT 192 82-83 GAAATTA GTGAGTGA

D4mit313 CTTCTTTAATCAA GGGCACATATGA 196 84-85 TCTCTGTCTCTGTG ACCTCCTG

D4mit344 CCAAACTCTTAGC ACACAGAAGAC 187 86-87 TTCTTCA ACTGAAGAAC

_7'J-Marker Forward Reverse Size, SEQ.
by ID

NO.

D4Mit51 CAGTTGTTAGAAG AGGTGCATATAC 123 88-89 CAGGATCCC CTGGGATACTC

D4Mit59 AGAGTTTGGTCTC TATCCAACACAT 108 90-91 TTCCCCTG TTATGTCTGCG

D4Mit59 GCCAGTGTGCTGA AGGGACCTGGA 119 92-93.

AAGACTG GACATCCTT

D4Nds16 CTGTAGGCTGCTT TGCCCCTTCAGC 94-95 TTATCTTTTG ACATGCCA

D4smh6b TGCAGTGTGACAT GGAAAGCCAGG 118 96-97 GTGCATAGAT CTACGCAGAA

D4smh6b CTGTAGGCTGCTT TGCCCCTTCAGC 102 98-99 TTATCTTTTG ACATGCCA

D4smh6b TAGTGTGGTTCCT CGGTCTACATAG 181 100-101 GACTAACCT TGAGTGATTC

D4smh6b AAA.AGCATCCTGC GGGTTATACAGA 83 102-103 ATCCTTCTG GAAACCCTGT

D4Xrf215 TTCCAAGCTCACA GTGCTGCTCTGC 124 104-105 CATCAGC ATTGAGTG

D4XrfZ43 GACAGTGTGGGAG CCCAAGGCATAG 203 106-107 AATCCGT GTCACAAT

D4Xrf243 ATTGTGACCTATG CGAAGGACCGTC 105 108-109 CCTTGGG ATCTGAGT

D4Xrf472 GGCTTTGATGTGA AGCTCCTCATCG 245 110-111 AAAAGGC CTCATGTT

D4Xrf472 TGGAACATCTCTG GGCTCTCATTGC 193 112-113 TCGGAAG CACCTTTA

AGACCTGC CTGGCAGA

_78_ Marker Forward Reverse Size, SEQ.
by ID

NO.

D4Xrf497 GCGAGACGAGTG ACACTGAAACCT 129 116-117 GGTAGTTC CGCTTGCT

D4Xrf497 AGCAAGCGAGGTT ACGGGGCTTGAT 204 118-119 TCAGTGT CCTTTTAT

Dshv4 AAGTTCATGGGCC TACTAGCTACCC 100-300120-121 TCACCACCTGTC TTCACATACC

DshvS ACCTAGCCACTGT ACAGAAGCAGC 100-300122-123 CTCAGTCT ATTTACACAG

Gnbl TGGGACAGCTTCC AATGGGAATTGT 213 124-125 TCAAGAT GCTCTTGG

Gnbl GGGCATCTGGCAA AGATAACCTGTG 281 126-127 AGATTTA TGTCCCGC

Gnbl GATGTCCGAGAAG TGTCAGCTTTGA 277 128-129 GGATGTG GTGCATCC

Gnbl ACATGCAGGCTGT TGTCAGCTTTGA 166 130-131 TTGACCT GTGCATCC

AAACCAA CCCTTAAA

ATGGCTC GTGCG

GAGATGAA TAAGAACC

k00954 GGTGAGAGTGGG AGGTTAGGCCCA 296 138-139 GAGATGAA ATTTCCTG

k00954 CCAGGGTTGCTGT CAGGTTAGGCCC 237 140-141 ACTGAGA AATTTCCT

CCTTCCC GGAAACCC

Maxker Forward Reverse Size, SEQ.
by m NO.

TGGAGGG AAACATCA

CCTTCCC GGAAACCC

ATGGGTG CGATATGT

CGCCATA TCTCACAC

CATAGG ATTTGCCC

CCAGATA ACAGCAAG

GCTCCAC TGTTTGGGG

TGTGAG TCCTGTGA

TTACCGA AATGACTT

TCAGAAGC TGTCCTGG

ACAGCCA TGTGACCA

GAGCAGG GAAAGGCA

GACCAAA GTGGGGGTG

Mmp23 CGGGTCCTCTCTT CTACATTTCCCT 297 170-171 CACCATA GAGCTGCC

-g~_ Marker Forward Reverse Size, SEQ.
by ID

NO.

Mmp23 GTTGACCATGTCG CCACCTCACGGA 111 172-173 GTAACCC AACTGAAT

Mmp23 GGTGTTTGGCTCA GATGCACACACA 197 174-175 CAAACCT AAAATCCG

Mmp23 ATCACCCACCAGA ACCCTCCAGGAG 255 176-177 ACGAAAA TAGGTGCT

GCAAGGT CCAAGCCA

CCTTGTT GCACAGAT

TGGAAAA CGATGCCT

TCCCCAA CAGACCGT

Pkcz TACAGCATATGCA ATTCCTCAGGGC 294 186-187 TGCCAGG ATTACACG

Pkcz GCAATCTCTTGTG ATTCCTCAGGGC 188 188-189 TCCAGGC ATTACACG

Pkcz TACAGCATATGCA GGCCTGGACACA 127 190-191 TGCCAGG AGAGATTG

Pkcz AAGTGGGTGGACA CAGCTTCCTCCA 201 192-193 GTGAAGG TCTTCTGG

Pkcz AGAGCCTCCAGTA TCGTGGACAAGC 297 194-195 GATGGCA TCCTTCTT

Pkcz CATCGAGTATGTC TTGTCCAGTTTT 156 196-197 AATGGCG AGGTCCCG

Pkcz CAGACTGGGTTTT GTCAAAGTTGTC 132 198-199 CCGACAT CAGGCCAT

Marker Forward Reverse Size, SEQ.
by ID

NO.

Pkcz AGGACGGACCCCA TGTCTCGCACTT 130 200-201 AGATG CCTCACAG

Pkcz CCAGAAGATGGA TCTACTGGAGGC 151 202-203 GGAAGCTG TCTTGGGA

Pkcz GAAAAACGACCA GATCTCAGCAGC 265 204-205 GATTTACG ATAGAACC

Pkcz ACACATTAAGCTG CAAACATAAGG 164 206-207 ACGGACT ACACCCAGT

Pkcz ACTGGGTGTCCTT CCTCTCTTTGGG 193 208-209 ATGTTTG ATCCTTAT

Pkcz GTCATAAAGAGGA GCTCTGTCTAGA 252 210-211 TCGACCA AGTGCCTG

Pkcz ACCAAGACCGAA GGCATTACACGC 223 212-213 GAGGGG TAACTTTTCC

TGGTAAG GGATGC

AATGTTT ACCAGGTT

GAGCAAT CCAGGTT

TGGGTTG CGCTTGGCT

GTAGGAACCTC CTGGC

RERl (1) GCCGATCCTGGTG ACAATGGCTCAA 224-225 ATGTACT AACCGTTC

RER1 (2) GCCTTGGGAATTT AGTACATCACCA 226-227 ACCACCT GGATCGGC

Marker Forward Reverse Size, SEQ.
by ID

NO.

RERl TAAAAGGCCATGC AGAGCTCTGTGG 228-229 GATAAGC GGTTCTCA

RERl GAAGGGGACAGT TCCATCAAGGAA 230-231 GTTGGAGA GGATCCAC

Tp73 GGTGGGTAATGAT TGACGTGGAGG 296-301232-233 TGGACT GAACTGCC

Tp73 TGAGATCTGGTGC GCCTGATCTAGG 222-229234-235 CCTCTCT CTGGAAAA

Txgpl AGGCAGAAAGCA CGACAGCACTTG 138 236-237 GACAAGGA TGACCACT

Txgpl CTGCAGATGTAGA CTGTGGTGGATT 269 238-239 CCAGGCA GGACAGTG

Txgpl TTGCCTAACACTC TATTAGGAGCAC 244 240-241 CCAAACC CACCAGGC

Txgpl ACCTGTCTTGTGG CTGTGGTGGATT 242-243 GTGGAAG GGACAGTG

ATTGACA GCCATTTT

GCTACCAA CATTGTGT

GGTAAATTGC TCCAGTTTG

TR1-likel(humaGCCACCATGCTGG TCACTCATGTTT 2520 250-251 n) GCCCTGCTGTCCT CCCCTGATTTCC

GGG

Tl-ike2(human)CTGATTTCCTGTG CATGCTGGCCTA 244 252-253 TTCCCGT CTTCATCA

Marker Forward Reverse Size, SEQ.
by ID

NO.

Tl-like3(human)GCCTTGCAGGTCA TCACTCATGTTT 2441 254-255 GCTACGGTGCTAG CCCCTGATTTCC

CAT

T1-like4(human)AGGAAGCAGAGA TCAGAACTGCCT 274 256-257 AAGGCCAG CTGAGCTG

T1-like5(human)TCTTCACGTACTG ACTACAGCATCA 175 258-259 GGGGAAC GCAGCAGG

T1-like6(human)AAGCTGAAGAACT TGGGCTACGACC 211 260-261 TCCCGGT TCTTTGAT

h-Trllike ATCTTCAGGCGCT GTACGACCTGAA 262-263 a CTGTCCT GCTGTGGG

h-Trllike ATCTTCAGGCGCT GTACGACCTGAA 264-265 b CTGTCC GCTGTGGG

h-Trllike ATCTTCAGGCGCT GAGTACGACCTG 266-267 c CTGTCC AAGCTGTGG

h-Trllike ATCTTCAGGCGCT TACGACCTGAAG 268-269 d CTGTCCT CTGTGGG

h-Trllike ATCTTCAGGCGCT TACGACCTGAAG 270-271 a CTGTCC CTGTGGG

h-Trllike GCTGTCCCGATGG ACCTTTTGTGGC 272-273 TGAAC CAGGATG

h-Trllike GCTGTCCCGATGG CACCTTTTGTGG 274-275 g TGAAC CCAGGAT

h-Trllike GCTGTCCCGATGG CCTTTTGTGGCC 276-277 h TGAAC AGGATG

h-Trllike CCTGAACCAGTGG ACCTTTTGTGGC 278-279 I

GCTGT CAGGATG

h-Trllike CCTGAACCAGTGG CACCTTTTGTGG 280-281 j GCTGT CCAGGAT

Marker Forward Reverse Size, SEQ.
by ID

NO.

h-Trllike TCATGTTTCCCCT CATGCTGGCCTA 282-283 k GATTTCC CTTCATCA

h-Trllike ATGAGCAGGTAAC TCATCACCTGGG 284-285 ACCTGGG TCTCCTTT

h-Trllike ATGAGCAGGTAAC TTCATCACCTGG 286-287 m ACCTGGG GTCTCCTT

mTrllike-lA TGGGTTGTGTTCT CCTTTTTACAGT 288-289 CTGGTTG CTGCCAGGT

mTrllike-1B TGGGTTGTGTTCT GATCCCCTTTTT 290-291 CTGGTTG ACAGTCTGC

mTrllike-2A ACGGGGTTGGTAC CACCCATTGTTA 292-293 TGTGTGT GTGCTGGA

mTrllike-2B ACGGGGTTGGTAC CACACACCCACC 294-295 TGTGTGT CATTGTTA

mTrllike-3A TGCATTGGCCAGA CGGCTGGGCTAT 296-297 CTAGAAA GACCTAT

mTrllike-3B TGCATTGGCCAGA CGGCTGGGCTAT 298-299 CTAGAAA GACCTATT

mTrllike-4A GTTCTGCAGCATG GGCAGTTGTGAC 300-301 ATGTCGT TCTGTTGC

mTrllike-4B GTTCTGCAGCATG CTGCAGGCAGTT 302-303 ATGTCGT GTGACTCT

mTrllike-SA CCATCCTTTTTGCC TCTGGAGGAACA 304-305 TGTCTT TGTGATGG

mTrllike-SB CACCATCCTTTTT GAACATGTGATG 306-307 GCCTGTC GGGCAAC

mTrllike-6A CAAAGCAGCAGG AAATGTACTGGC 308-309 AGGAGTG CAGGCAAC

Marker Forward Reverse Size, SEQ.
by ID

NO.

mTrllike-6B AGTGCTAGACCCA AAATGTACTGGC 310-311 GCACCAG CAGGCAAC

mTrllike-7A GCACTGACCAGTC GTCCCCAGAGAA 312-313 TGTCACC AAGCACAG

mTrllike-7B CAGTCTGTCACCA CAGTGGTCCCCA 314-315 CCTCTGG GAGAAAAG

mTrllike-8A TACTATTCGGGGC GCAGCACTATGT 316-317 TTGTTGG GCCTGGTA

mTrllike-8B TACTATTCGGGGC GCCTGGTATTTG 318-319 TTGTTGG ATCGCTTT

mTrllike-9A GCTCAGCTAGGGA CAGCTCAGGGAC 320-321 TGGAGAA ACAATGAA

mTrllike-9B TCCTACAGGCTAG CAGCTCAGGGAC 322-323 GGCTCAG ACAATGAA

mTrllike-l0AGGGACTGATGTGT AGGCGTCCCAGG 324-325 GGCTTGT AATAGAAG

mTrllike-lOBGGACTGATGTGTG AGGCGTCCCAGG 326-327 GCTTGTTT AATAGAAG

mTrllike-11ATGTTTCTGTTCTGG ATCTGCAGGCAG 328-329 TGGCTG GATCAGAC

mTrllike-11BCTCAGTGGTGGGT ATCTGCAGGCAG 330-331 GACAGTG GATCAGAC

Mutationl ACACACAGTACCA CCTGTGGTGATC 182 332-333 (mouse) ACCCCGT AAGAAGCA

Mutation2 TGCTTCTTGATCA GCAACAGAGTC 131 334-335 (mouse) CCACAGG ACAACTGCC

Mutationsl+2ACACACAGTACCA GCAACAGAGTC 293 336-337 (mouse) ACCCCGT ACAACTGCC

Marker Forward Reverse Size, SEQ.
by ID

NO.

34m15-T7 GGGTTTATGTGGC ACTCCATTTGCC 118 338-339 AAGCACT TTTTGTGG

34m15-SP6 CGCTACTTCGCTT ATGATGACGTAC 150 340-341 TTATCCG GACGACGA

GGAGAAGTC ACGCCAGG

37D20-T7(3)*AGTGAGAGGCCCA GATCTGATGCCC 247 344-345 GTCTCAA TCTTCTGC

CCAACAC TTACCCAG

GTCTGTC CTCTTCTA

CCAGCCC CACTGCTC

GGTCTTC GCTCCTGC

GCCGCC TCCCCAAG

TAATCTC AGAAAGCA

AAACTCACA AAGAAAAAGTC

AGACA

CCTCAAA TCAGCTAT

AATAAGT CTCCTTGAG

130A12-T7* GCTCAGTGGGTAA CTACCCTGCCGC 242 364-365 GAGCACC TAATCTCA

_87_ Marker Forward Reverse Size, SEQ.
by ID

NO.

ACCCTAA TTCACCTG

ACGGGGA CAAGCTCC

GTTTCGG AGGCCAGG

GTTTCGG AGGCCAGG

147a15-T7* GTGGTTGCTGGGA CAAGCAACCAA 101 374-375 TTTGAAC ACAACCAAA

AAATCTG CATTCCCT

GCACAGG ATGTGATT

GCAGTTG CATATCCA

CCGCTT GCCTTGAA

CAGGAAG ATGAATAA

TCTGTTCA TGGCAAGG

TGGCCTA TTTCCTCGT

CTCTTTG CCAGAC

TTTACATGG AAAAGGAT

_88_ Marker Forward Reverse Size, SEQ.
by ID

NO.

189M12SP6* CCGTCGACATTTA GATAGTGGGGTG 189 394-395 GGTGACA GTGGGTAA

GGTGACA GTAACTGA

22764-T7'~ GGAAGCAAATGCT TATCCCTAGCCC 243 398-399 CCACTAAA CTTGTGTG

GGTGACA TAGTGACC

TCATTGG CTGCTTACA

TGTGACT ACGGCTGG

TCCCCTG CCTCTCAG

280612-T7* GGGCTGGGAATTG TGAATCCCTTAC 420 408-409 AACCTAT AGCCTTGC

CACTCCT TAGAAGAT

TAGGCAA TGTCCTCA

284D21-SP6 ACAGAAATCCCTC TCAGTGTGGACC lOS 414-41S

ATGCGA AGAAAGTCC

TCTTGATAA CAAGCTGTCTG

298e4-T7(3)*TCTCCCCTTTTACC GCAAGGAGTCA 180 418-419 ACTCCC AAAACAGCA

CAAACCA CAACTCCA

Marker ~ Forward Reverse Size, SEQ.
by ID

NO.

GGGACGA GCAATCCAC

CACCCTA TTCACCTG

TGCTGAT TGGAGCTT

350D2-T7* TGTAGGGAATGTT ACATGGAACAG 295 428-429 TCTGCACC GATTCTGGC

ACAGACAA TTACTGAC

TGTGGGT TTGAGGCA

CAGCCTCAG AAGCCTGG

CTCTTGT AGTGGTGGG

382A8-SP6(2)CAGCGATTGGCTC GGGGCTTCCTTT 531 438-439 TTCTCTT CTGAGGTA

CTTGGTA TGCTTCTC

GGTTATC AATGGAAAG

CACTCCT CCCAAACC

GGGACGA GCAATCCAC

TGGTCTC TTCTGCCT

Marker Forward Reverse Size, SEQ.
by ID

NO.

CTATTCCTTTG GGCTTTTTGATA

CA

GTTGACC TCAGGGCA

TTTTGTT AGGTTCCA

GCTGCTG AGGTTCCA

AACTCAGGA CACCTCCA

GTTGTTG TGAATGAT

CTTGGTA TCCCCTCC

TGGACAA CCAGGAAG

AATCTGA GCCTGAGC

GTTTGGACA ATCCCAGC

CCCTCA GAGGTCCA

D4Mon1 AGGCCTACCCAAG GCAGTGAGCTGC 201 472-473 GACATCT AGAGTTTG

D4Mon2 AGACACCCTAGGT TGATCTTTCCAA 151 474-475 CCTGCTG ACGCATAAGA

D4Mon3 GCAAGCAACCTGA GCTTACGATGGT 188 476-477 ACATGAA CGTGAGGT

Marker Forward Reverse Size, SEQ.
by ID

NO.

D4Mon4 ACATGCCTGCCTA GGAACCTGTTTT 197 478-479 TCTTTGC CCATGGTG

D4Mon5 ACCTTGTTCCTGG TAGCTGGGACGT 200 480-481 TGTGAGC GGTATGGT

D4Mon6 CCATGGGAGACCA TGAGTGTCCTCT 206 482-483 GAAGGTA GCCTGATG

D4Mon7 GCGCTGACATCCT CCCACTATGGTC 187 484-485 CCTATGT CCAGAGAA

D4Mon8 . TTGCACGTCTTTG AAAGGGGAATA 219 486-487 TTTCGAG GACCTGAGTAG

AA

D4Mon9 CCAAGAGTCAGCC GGACAGGTAGCT 200 488-489 TTGGAGT CACCCAAC

TrllikeulcDNATGCCAGCTTTGGC TTCATTGTGTCC 490-491 mouse TATCAT CTGAGCTG

Trllikeu2cDNAAGCTTTGGCTATC ACCACCGCCACT 492-493 mouse ATGGGTCTCAG GTTCTCATCT

Trllike A1 TGTGGGGGAAGA TGATGTGTGGCT 5935 494-495 (mouse) ACATAGAA TGTTTCTCTT

Trllike A2 ATAGGTGGGGAG TGATGTGTGGCT 5903 496-49?

(mouse) GGAGCTAA TGTTTCTCTT

TRl like-2 TGTGCCTGTCACA CATGCTAGCACC 498-499 (human) GCAACTT GTAGCTGA

TRl like-3 GGAGACCTTCCCC GCTGTAGTTGAA 500-501 (human) TCCTTCT GAGGGCGT

TRllike-4 GTGCTTGGCTTCC CAGGTCGTACTC 502-503 (human) TCCAG CATGTCCA

TRl like-5 TGGAGTACGACCT ACTCATCCTGGC 504-505 (human) GAAGCTG CACAAAAG

Marker Forward Reverse Size, SEQ.
by ID

NO.

TRllike-6 GAACAGGAGGAC CTTTTGTGGCCA 506-507 (human) GCTGAGG GGATGAGT

TRl like-7 TCACCTCACCTGG GTACGACCTGAA 508-509 (human) TTGTCAG GCTGTGGG

TRl like-8 GGCTGAGATCACA CCGTGCCTGTTG 510-511 (human) GGGTTGGGTCACT GAAGTTGCCTCT

C GCC

118e21-0 AATTCCCAGCAAC CAGACACTCCAG 585 512-513 CACTCAC AAGAGGGC

118e21-1 TGACTGCTCTTCC TTTGTGGAATAG 588 514-515 GAAGGTT CCAAAGCC

TCCCCC CACCTTAT

118e21-3 TAGGAGTGCCCCA TCATTGTACCCA 518 518-519 TAGGTTG GCCAGTCA

118e21-4 AGGACTGAGCCTG CTGGGCGTTTTG 552 520-521 GATGAGA TTTTGTTT

118e21-5 CTTCCTCCTGCAG ACCCTGCTACAA 546 522-523 CTACCAC CGCAGACT

118e21-6 TCCAACCTTGACA AGCCAGGGCTAC 584 524-525 CCCATTT ACAGAGAA

139J18T7(1) CTGCTTTTCCTCA ATTCGCCGTTAG 526-527 GCAACTG AAGCTAGG

139J18T7(2) AACTGTACGTGGC ATTCGCGGTTAG 528-529 TGCTGGT AAGCTAGG

Agrin(CA)n GCCAGGTGACCCT GAGAGATGGCA 271 530-531 TATGAAA GACAGAGGC

Agrin(TG)n AGCTCTCTGTCCC TGCCAACCACTA 157 532-533 TGGTGAA GCCTCTCT

Marker Forward Reverse Size, SEQ.
by ID

NO.

repeatl CTGAACCCTCCAC AGCCAGGGCTAC 205 534-535 TCTCCTG ACAGAGAA

repeat2 AGCCAGGGCTACA ACCCTGCTACAA 153 536-537 CAGAGAA CGCAGACT

repeat3 GCAAGTTTCAGGA CCCCAGAACCAG 166 538-539 GCTAGGG AGACCATA

repeat4 CTAGGGGACTCTG CA.AGACACCCA 195 540-541 CCAAGTG GTCCCAACT

repeats TACTTCCCCTTTCC TCCTTGGTGCTT 232 542-543 CGAACT ACCCTCAC

repeat6 TGTTCCTGAGTTC ATTCCCAGCAAC 269 544-545 ACAACGC TACATGGC

repeat? ACATGTCCACTGT TGTCATGAGTTT 246 546-547 GGCAAAA GAGGCCAG

repeat8 ATCAGACAGCCCA TATGTGCCACCA 206 548-549 CAACCTC CACCTGTC

repeat9 GCTCAAGGAAGG TGCTCTTAACAT 201 550-551 ACACACCT TTTGAGCCAT

repeatl0 GCTCAGCCCCTGA GGGATCTGCCTG 111 552-553 ATCAATA TCTTACCA

repeatll GGAAGGTAGGGC GCTCCAAGATCT 277 554-555 CTGGTAAT GTGCGATT

repeatl2 TTAGCGTTAGGGT GGAGACTACGG 150 556-557 GAGGGTG ACTTGTGGC

repeatl3 CAGTTCTTCCCGA TTTCTGGGAACT 174 558-559 AAACCAC GAGATGGC

repeatl4 GTTGGGGCTGCTC GCTGTGGCTCTC 422 560-561 ATAGAAA TTGGAGTT

Marker Forward Reverse Size, SEQ.
by ID

NO.

repeatl5 CTCTGATTTCCCA AAGAGGGAGCA 152 562-563 CATGCCT CTGAGGACA

repeatl6 CAGCAGCAAATGA GAGGCAGGCAG 147 564-565 CCTTTCA ATTTCTGAG

repeatl7 GTTTCACATGTTG GGGACCTTTGGG 131 566-567 TGGTGGC ATAGCATT

repeatl8 TCAGACATCTCTG TTCACTAAGTTG 160 568-569 GCCTCCT CCCAGGCT

repeatl9 TGCCTTTTTCTCAC TTAGAAGCAGA 250 570-571 ATTGTCTC GGCAGAGGC

repeat20 GACCTTTGGAAGA TGGCAGCTCACA 296 572-573 GCAGTCG ATGTCTTT

GGAAGAA ACACAAACAG

GGAGAAT GGTACCGAGCTG

TGTCAGG ACCAAGT

TCTTCTG' TCATACAC

GGGTTCT GGTTAAATGC

GAGGACT GAAAGAACTTG

GA

GCGCTTG GGGGACAGG

AGCTGTT CTGAGAAT

Marker Forward Reverse Size, SEQ.
by ID

NO.

GTTGTTC GGTGAAG

GTCTGAA TACCTTTTT

TAGCAA CCCAGTAA

GATCAGA CGGCATAA

AGCCTCAACC AGACTGTGAGA

CAGAGA AAGAGCAG

ATCCATA GCAAAATGT

GTTCAGG CAGGCAAC

GTTCCTTT GAACGATAG

TGTGTTT TGGAGAAAGA

AGCAGAG ACCTAGCA

GACCTC CCATGTTA

TTGTGTG TTGTTCCT

ACCATCT TGCAGCAT

Marker Forward Reverse Size, SEQ.
by m NO.

GCTTTGT TCTCTCTA

TTTGTTT GGGTTCTA

TGACTTGC CAAAGCCTA

AGGGAAG AGCTCTCC

CTAGGTTT CACAGGAG

AGGCAGAA GGAATACA

SHANRU29 AAAGGGATGTGTG CAAA.ACCCAACC 195 630-631 GCGTAAG TTCTCAGC

ATAGAGG GGCTGTCT

CGCACTT AAGAGGCTA

GGGGTAA TGTCTCTT

TGCTTCT CGTCAGAT

ATATTCA CCAAAAAGC

CTCAACT AACAATTGAA

CACAGTGAGA TGACAGCA

Marker Forward Reverse Size, SEQ.
by ID

NO.

GAGACCA GGACTCATT

CTCTCCTCTGA GAAGTAGGC

CCTCTTC AAAAGGAA

Although the foregoing invention has been described in some detail by way of illustrating and example for purposes of clarity of understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims.
All publications, patents, and web sites are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent, or web site was specifically and individually indicated to be incorporated by reference in its entirety.

SEQUENCE LISTING
<110> Bachmanov, Alexander A
Beauchamp, Gary K. a Chatterjee, Aurobindo De Jong, Pieter J.
Li, Shanru ;
Li, Xia Ohmen, Jeffrey D
Reed, Danielle R.
Ross, David Tordoff, Michael G.
<120> GENE AND SEQUENCE VARIATION ASSOCIATED WITH SENSING
CARBOHYDRATE COMPOUNDS AND OTHER SWEETNERS
<130> Gene & Sequence Variation......
<140>
<141>
<150> 60/200,794 <151> 2000-04-28 <160> 652 <170> PatentIn Ver. 2.1 <210> 1 <211> 2577 <212> DNA
<213> Mouse <400> 1 atgccagctt tggctatcat gggtctcagc ctggctgctt tcctggagct tgggatgggg 60 gcctctttgt gtctgtcaca gcaattcaag gcacaagggg actacatact gggcgggcta 120 tttcccctgg gctcaaccga ggaggccact ctcaaccaga gaacacaacc caacagcatc 180 ccgtgcaaca ggttctcacc ccttggtttg ttcctggcca tggctatgaa gatggctgtg 240 gaggagatca acaatggatc tgccttgctc cctgggctgc ggctgggcta tgacctattt 300 gacacatgct ccgagccagt ggtcaccatg aaatccagtc tcatgttcct ggccaaggtg 360 ggcagtcaaa gcattgctgc ctactgcaac tacacacagt accaaccccg tgtgctggct 420 gtcatcggcc cccactcatc agagcttgcc ctcattacag gcaagttctt cagcttcttc 480 ctcatgccac aggtcagcta tagtgccagc atggatcggc taagtgaccg ggaaacgttt 540 ccatccttct tccgcacagt gcccagtgac cgggtgcagc tgcaggcagt tgtgactctg 600 ttgcagaact tcagctggaa ctgggtggcc gccttaggga gtgatgatga ctatggccgg 660 gaaggtctga gcatcttttc tagtctggcc aatgcacgag gtatctgcat cgcacatgag 720 ggcctggtgc cacaacatga cactagtggc caacagttgg gcaaggtgct ggatgtacta 780 cgccaagtga accaaagtaa agtacaagtg gtggtgctgt ttgcctctgc ccgtgctgtc 840 tactcccttt ttagttacag catccatcat ggcctctcac ccaaggtatg ggtggccagt 900 gagtcttggc tgacatctga cctggtcatg acacttccca atattgcccg tgtgggcact 960 gtgcttgggt ttttgcagcg gggtgcccta ctgcctgaat tttcccatta tgtggagact 1020 caccttgccc tggccgctga cccagcattc tgtgcctcac tgaatgcgga gttggatctg 1080 gaggaacatg tgatggggca acgctgtcca cggtgtgacg acatcatgct gcagaaccta 1140 tcatctgggc tgttgcagaa cctatcagct gggcaattgc accaccaaat atttgcaacc 1200 tatgcagctg tgtacagtgt ggctcaagcc cttcacaaca ccctacagtg caatgtctca 1260 cattgccacg tatcagaaca tgttctaccc tggcagctcc tggagaacat gtacaatatg 1320 agtttccatg ctcgagactt gacactacag tttgatgctg aagggaatgt agacatggaa 1380 tatgacctga agatgtgggt gtggcagagc cctacacctg tattacatac tgtgggcacc 1440 ttcaacggca cccttcagct gcagcagtct aaaatgtact ggccaggcaa ccaggtgcca 1500 gtctcccagt gttcccgcca gtgcaaagat ggccaggttc gccgagtaaa gggctttcat 1560 tcctgctgct atgactgcgt ggactgcaag gcgggcagct accggaagca tccagatgac 1620 ttcacctgta ctccatgtaa ccaggaccag tggtccccag agaaaagcac agcctgctta 1680 cctcgcaggc ccaagtttct ggcttggggg gagccagttg tgctgtcact cctcctgctg 1740 ctttgcctgg tgctgggtct agcactggct gctctggggc tctctgtcca ccactgggac 1800 agccctcttg tccaggcctc aggtggctca cagttctgct ttggcctgat ctgcctaggc 1860 ctcttctgcc tcagtgtcct tctgttccca gggcggccaa gctctgccag ctgccttgca 1920 caacaaccaa tggctcacct ccctctcaca ggctgcctga gcacactctt cctgcaagca 1980 gctgagacct ttgtggagtc tgagctgcca ctgagctggg caaactggct atgcagctac 2040 cttcggggac tctgggcctg gctagtggta ctgttggcca cttttgtgga ggcagcacta 2100 tgtgcctggt atttgatcgc tttcccacca gaggtggtga cagactggtc agtgctgccc 2160 acagaggtac tggagcactg ccacgtgcgt tcctgggtca gcctgggctt ggtgcacatc 2220 accaatgcaa tgttagcttt cctctgcttt ctgggcactt tcctggtaca gagccagcct 2280 ggccgctaca accgtgcccg tggtctcacc ttcgccatgc tagcttattt catcacctgg 2340 gtctcttttg tgcccctcct ggccaatgtg caggtggcct accagccagc tgtgcagatg 2400 ggtgctatcc tagtctgtgc cctgggcatc ctggtcacct tccacctgcc caagtgctat 2460 gtgcttcttt ggctgccaaa gctcaacacc caggagttct tcctgggaag gaatgccaag 2520 aaagcagcag atgagaacag tggcggtggt gaggcagctc agggacacaa tgaatga 2577 <210> 2 <211> 11809 <212> DNA
<213> Mouse <400> 2 atctgagcct tagacacagc actggtgcca ggcaaacact cctgggccta catgcttggg 60 gcctcttcat attccaaaag ctgtctttgg gtaagatgaa gttcctctgg cagtggcatg 120 agtgctgaag gctctttccc tgcccttcac ctgctttctt gatagtctct ctgcatacca 180 aacaggccct tgtctcctgg gaaatggaaa ctatgaaatc aatagctgag gcttctctag 240 gaaagcctgc cctggtcagt acaacctgtt tcacagcttc tatagaatag ttacatcagc 300 cttctgaaga tggcctctta gagcacatgc acccccaaga ttctaagatg tcaatactaa 360 ctgaccaaac catacctctc tagccagccc tgctgctcct gttgtctggt acccaggtga 420 ctgaggacat gactggtgga aggaaactag gcccctttgt ctgtcagatg gccataccca 480 gcatggctga tgcccagtgt ataagaccct acgcttttcc actggtctta atgttaaacc 540 ctaggacagt gtcctcagca tagctggtgt gtgtgaatgc aaactttggg gcatatctct 600 tccattaagc actgtgatat atgtagtatt tccaacaaat aaattatacc tacatgattg 660 ggtatagcat tctgggatgg gtcacaagtg tgtcaggtgc ctaattatgt gggggaagaa 720 catagaaata tataggtggg gagggagcta accctaggaa taaggctaaa gcatgtgtct 780 ccagtcctga agactcaaag ggcaacgtga atcatgagac atgttcagga ctgaaggagt 840 tgccatgtat ctgtccttga tgtatcttaa tcatacatac actatgagat ctgtgttacc 900 tccattttgc aggtgagaaa agaaacacct gaatggccta ccttaaaggg ctaagtggga 960 aaataggtct gaagataacc caggcactgt gtgacaaagc gggaagaaaa ctagagatgc 1020 tttcttcatg gcaacaacct agagggtaca acctagtggt ttcttcttgg tactccactg 1080 tatacacccc atctgcttgg gctgtacatt gtctgaccat gcttataaca aaagtcacat 1140 actactagcc aagactgaga acttagagcg actggccaga aagtaaagat acaacagttg 1200 atatgtgtgc cacacacaga tccatgtgta catgtctatt aattatgtga acgtgctttg 1260 tggacatcct cacaaagcag cagggaaatg caaaggtcat ttccataaca cctgctggac 1320 accatatgac attgagatta ccggggtgcc cattccaaca agagttaata gctcccccta 1380 tgtttgggtg ccagaaacct gatttgttag caatagctcc ctcacatcca gattaagagg 1440 gggatggctt agctagggtt actatgatga aactatgacc aaagcaactt gtgggtaaaa 1500 gggtgtattt ggcttacact tccatatcac ttcatcaaag tgaggacagg aactcaaata 1560 gagtaggaat ttggtgacaa gagctgatgt agaggcaatg cagtggtgcc acttagtggc 1620 gcgctcagtc tgctcccttt cttaatagaa tgcaagacca ccagcccatg ggtggcacca 1680 caatgggacc gggcccttcc ccatcggtca ctaagaaaat gccctacagc cagatcttat 1740 ggagacattt tctcaacgga ggctcactcc tttcagataa ctctatatca aattgacata 1800 aaccagaaca gaggaggagg ctaagaagga aactgccaat tgcatacatg cacacacctg 1860 gccctagcag ctgcaggaag ctatttgttt atggcctttt ctcattttca tggaccagca 1920 tgagcactct gcagagagag atgcctgcat gcctgccaag gcaggagtgc ttacactgaa 1980 ggtcaacagg atggcagggg ggctgcagag cttccaagtg tcagaacccc agcagaagag 2040 ctgagaccct tgcccgagga ctcaggcggg ttgggaaggc caggaaattc agccagagct 2100 cttcttcaga tggggtacca tctgaaggtt agaccagcta gccagctgtt gttgagggac 2160 cacctctgca gcccctacct ttggaagata gaaagtgtct ctgtgacaag tatggccatt 2220 gtgccccctt attccacagt caacagaaac cctggaatcc tgaacacttc tgcagcttct 2280 tttttacagt ctgccaggtt gctctaggaa tgaagggtgc cgagaggctt gggcgtaggc 2340 aggtgacaag accacagtta gtggtcacag ctggcttact ggatcactct tggacagagt 2400 ttgttagata tggagtggag tatacacaag gcatcaggcg ggggatattg aatgtatcac 2460 cggagctcct tggggcttgg cagccaagca cagcagtggt tttgctaaac aaatccacgg 2520 ttccctcccc ttgacgcagt acatctgtgg ctccaacccc acacacccac ccattgttag 2580 tgctggagac4ttctacctac catgccagct ttggctatca tgggtctcag cctggctgct 2640 ttcctggagc ttgggatggg ggcctctttg tgtctgtcac agcaattcaa ggcacaaggg 2700 gactacatac tgggcgggct atttcccctg ggctcaaccg aggaggccac tctcaaccag 2760 agaacacaac ccaacagcat cccgtgcaac aggtatggag gctagtagct ggggtgggag 2820 tgaaccgaag cttggcagct ttggctccgt ggtactacca atctgggaag aggtggtgat 2880 cagtttccat gtggcctcag gttctcaccc cttggtttgt tcctggccat ggctatgaag 2940 atggctgtgg aggagatcaa caatggatct gccttgctcc ctgggctgcg gctgggctat 3000 gacctatttg acacatgctc cgagccagtg gtcaccatga aatccagtct catgttcctg 3060 gccaaggtgg gcagtcaaag cattgctgcc tactgcaact acacacagta ccaaccccgt 3120 gtgctggctg tcatcggccc ccactcatca gagcttgccc tcattacagg caagttcttc 3180 agcttcttcc tcatgccaca ggtgagccca cttcctttgt gttctcaacc gattgcaccc 3240 attgagctct catatcagaa agtgcttctt gatcaccaca ggtcagctat agtgccagca 3300 tggatcggct aagtgaccgg gaaacgtttc catccttctt ccgcacagtg cccagtgacc 3360 gggtgcagct gcaggcagtt gtgactctgt tgcagaactt cagctggaac tgggtggccg 3420 ccttagggag tgatgatgac tatggccggg aaggtctgag catcttttct agtctggcca 3480 atgcacgagg tatctgcatc gcacatgagg gcctggtgcc acaacatgac actagtggcc 3540 aacagttggg caaggtgctg gatgtactac gccaagtgaa ccaaagtaaa gtacaagtgg 3600 tggtgctgtt tgcctctgcc cgtgctgtct actccctttt tagttacagc atccatcatg 3660 gcctctcacc caaggtatgg gtggccagtg agtcttggct gacatctgac ctggtcatga 3720 cacttcccaa tattgcccgt gtgggcactg tgcttgggtt tttgcagcgg ggtgccctac 3780 tgcctgaatt ttcccattat gtggagactc accttgccct ggccgctgac ccagcattct 3840 gtgcctcact gaatgcggag ttggatctgg aggaacatgt gatggggcaa cgctgtccac 3900 ggtgtgacga catcatgctg cagaacctat catctgggct gttgcagaac ctatcagctg 3960 ggcaattgca ccaccaaata tttgcaacct atgcagctgt gtacagtgtg gctcaagccc 4020 ttcacaacac cctacagtgc aatgtctcac attgccacgt atcagaacat gttctaccct 4080 ggcaggtaag ggtagggttt tttgctgggt tttgcctgct cctgcaggaa cactgaacca 4140 ggcagagcca aatcttgttg tgactggaga ggccttaccc tgactccact ccacagctcc 4200 tggagaacat gtacaatatg agtttccatg ctcgagactt gacactacag tttgatgctg 4260 aagggaatgt agacatggaa tatgacctga agatgtgggt gtggcagagc cctacacctg 4320 tattacatac tgtgggcacc ttcaacggca cccttcagct gcagcagtct aaaatgtact 4380 ggccaggcaa ccaggtaagg acaagacagg caaaaaggat ggtgggtaga agcttgtcgg 4440 tcttgggcca gtgctagcca aggggaggcc taacccaagg ctccatgtac aggtgccagt 4500 ctcccagtgt tcccgccagt gcaaagatgg ccaggttcgc cgagtaaagg gctttcattc 4560 ctgctgctat gactgcgtgg actgcaaggc gggcagctac cggaagcatc caggtgaacc 4620 gtcttcccta gacagtctgc acagccgggc tagggggcag aagcattcaa gtctggcaag 4680 cgccctcccg cggggctaat gtggagacag ttactgtggg ggctggctgg ggaggtcggt 4740 ctcccatcag cagaccccac attacttttc ttccttccat cactacagat gacttcacct 4800 gtactccatg taaccaggac cagtggtccc cagagaaaag cacagcctgc ttacctcgca 4860 ggcccaagtt tctggcttgg ggggagccag ttgtgctgtc actcctcctg ctgctttgcc 4920 tggtgctggg tctagcactg gctgctctgg ggctctctgt ccaccactgg gacagccctc 4980 ttgtccaggc ctcaggtggc tcacagttct gctttggcct gatctgccta ggcctcttct 5040 gcctcagtgt ccttctgttc ccagggcggc caagctctgc cagctgcctt gcacaacaac 5100 caatggctca cctccctctc acaggctgcc tgagcacact cttcctgcaa gcagctgaga 5160 cctttgtgga gtctgagctg ccactgagct gggcaaactg gctatgcagc taccttcggg 5220 gactctgggc ctggctagtg gtactgttgg ccacttttgt ggaggcagca ctatgtgcct 5280 ggtatttgat cgctttccca ccagaggtgg tgacagactg gtcagtgctg cccacagagg 5340 tactggagca ctgccacgtg cgttcctggg tcagcctggg cttggtgcac atcaccaatg 5400 caatgttagc tttcctctgc tttctgggca ctttcctggt acagagccag cctggccgct 5460 acaaccgtgc ccgtggtctc accttcgcca tgctagctta tttcatcacc tgggtctctt 5520 ttgtgcccct cctggccaat gtgcaggtgg cctaccagcc agctgtgcag atgggtgcta 5580 tcctagtctg tgccctgggc atcctggtca ccttccacct gcccaagtgc tatgtgcttc 5640 tttggctgcc aaagctcaac acccaggagt.tcttcctggg aaggaatgcc aagaaagcag 5700 cagatgagaa cagtggcggt ggtgaggcag ctcagggaca caatgaatga ccactgaccc 5760 gtgaccttcc ctttagggaa cctagcccta ccagaaatct cctaagccaa caagccccga 5820 atagtacctc agcctgagac gtgagacact taactataga cttggactcc actgacctta 5880 gcctcacagt gaccccttcc ccaaaccccc aaggcctgca gtgcacaaga tggaccctat 5940 gagcccacct atcctttcaa agcaagatta tccttgatcc tattatgccc acctaaggcc 6000 tgcccaggtg acccacaaaa ggttctttgg gacttcatag ccatactttg aattcagaaa 6060 ttccccaggc agaccatggg agaccagaag gtactgcttg cctgaacatg cccagccctg 6120 agccctcact cagcaccctg tccaggcgtc ccaggaatag aaggctgggc atgtatgtgt 6180 gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtat gtacgtatgt atgtatgtat 6240 caggacagaa caagaaagac atcaggcaga ggacactcag gaggtaggca acatccagcc 6300 ttctccatcc ctagctgagc cctagcctgt aggagagaac caggtcgccg ccagcacctt 6360 ggacagatca cacacagggt gcgggtcagc accacggcca gcgccagcca cgcgggaccc 6420 ctggaatcag cttctagtac caaggacaga aaagttgccg caaggcccct tactggccag 6480 caccagggac agagccacat gcctaagcgg caagggacaa gagcatcgtc catctgcagg 6540 caggatcaga cccgggtcag ttctggactg gcccccacac ctgaatcccg gagcagctca 6600 gctggagaaa agagaaacaa gccacacatc agtcccataa aattaaacgc tttttttagt 6660 gtttaaaata gcatttacac agaagcagca tttacacaga agcagctcta tgtcaactac 6720 ccagtcactc agactttgac acagtgtcta gtgtagatgt gtggggccgc tgtgccggga 6780 tggcagtggc acatgatgat gggcagccac cagaacagaa acagaacagg gcccagctct 6840 gcagctcttg tgttcactgt cacccaccac tgagactgag acagtggcta ggtgccaggt 6900 ctctctcctg tctctcctac tagctaccct tcacatacct tcagtacaaa ctgtgttgtc 6960 atgtgccaag tagcaggtgg ggaaaggggc atgcaaactg cccctttggg taactagctg 7020 ccacccttag agcaggcagg ctagcaataa ataaataagt tagaccccac ctgggcagcc 7080 agagaggttt gaaggctctg tctaacccct caaaaatccc accttggcct gacaggtgag 7140 gcccatgaac ttagcgacag tcagcctgtg tccctgtgca cagttctgtg aggctttggg 7200 gcaaggggta ccaagagccc aagagagcct ttcttgttct aaatggaggt cacttccaaa 7260 gaagggaacc aggaggtggt ccctgagact tg~gctgagg acttaaagtc agagatgtct 7320 ccttacaaga ctctatagat acttgagctg taccaccatc agcagcccca agagcagaca 7380 aaatgtcaag ccaatatcct ggtggtatgg ctgccctcag gccctcctct gtagcctgct 7440 ccctctgccc tggcccagag cccacagctg atctatcctg gctggccacc accacggcca 7500 gcgcagagct cctggcacag caggagcaca gactcagcca caggcagcgc tgaagacatt 7560 ggttgatcat cacatgatgt ccacaaagaa ctcacagggg tttcccatgg ccttttggaa 7620 ggactggcgg ctacctgtaa gttctggagg gacagcagcc agctcccgga cgggtggccc 7680 tccaggtggc ccacccacta ctgcataggc ctttgtaagg gggtgcagtg gggggagccc 7740 tggggcaaca gctgaagcct gacttcgagg gctactgcca cggctaagct ggctgacagg 7800 ccgctcccac cagccggtgc taccagaccc acttggtact gtgtggtctg attcactgcc 7860 actaccccca gctccagttg cccggcgctc ctctcggcct ggggtccgat ggctgctccg 7920 tgtggaccca ctgctcttgc tccctagggg gagggaaggg gacaacagag tcagcacgag 7980 gcctggccac ttccagggcc accagctgct cccagacagt cagggcagga cctggtaagc 8040 ctggagatgg taggggaatg gcagccatgc agataccagg aacagctgag aggcgagaag 8100 ctaggggcag tggcagacag cagggacaac aggggccagc ctggcacccc acacctaacc 8160 ccaatgcttg aaccaagggt taatgttaca gctgagaaac taaaaaccag cgaaggccct~8220 gtgtgcccag cattcccatt agccatcctg ggttcaccac ccaaagaccc aaccagggtc 8280 cacccaaccc caggaccctg gtcatctaat ttgcttagcc cctgtcctga aagtagtggg 8340 aacctgaaaa cacgtgctgg ctggggacat gctgagaggg acacaggggg acctggctta 8400 ccggcccgag agtccactct gctagtcctt cagtctaagg cttgctcagc acaaagcaag 8460 ggatagcaca agtcacacac cagtccagtg ctcaccaatg gctaatagga cgattttggg 8520 ccaagctgag cctgggtaca tgcaagggcc tgtccatggt caggattcac tcgatagctt 8580 ccccttgggc tttgccaccc tctggcccaa cctctcctga gtctttctct ggaccttgta 8640 gcacaagtgt gccccactct gcctaagacc tccacatcag tccatctcct cctgagggac 8700 acccaccctt caagatcttc aatatccctg ggatatgctt taacactgat atgctttaac 8760 agtgttgctt gatactctta tctggcactc tgttgggatg caggctccat aactgataaa 8820 gcccattctc cccctagctt ggggcctaga gagtgcccct acctgctatc agtggttact 8880 ttcattcttg ccatatcatc tcctggcctc ttgcctctgc cacctagcac accaggctgt 8940 cttcctattc tctaacggct tctacccaca tcagcccctc cctgtcccac acactgactc 9000 ttgagatgga acccaccggg actcaaacac acagcaggag cacagaggga agcgtcgggg 9060 ccaggcagag cgtgggagtg ggagggagtg ggaggagggg tggcacgcct ctcaccttca 9120 ctctgctggc tcccagcact gccgctgccg cagctgaagc cagggtcctg gtaagcaggc 9180 gggaagcagg gcgggggtcc tgggtactgg taggggtagc cttgacccaa gggccagggt 9240 actgatgggt ggggcagtgg ggccagtgtg tcctgatctg aggctccact ggagccactg 9300 ttgaggttca gggatgcgag gtctggcagg gagggaggga gggaggggta agtgaaggca 9360 aatgaatgag gccacagcaa ccctacccaa ccgcacccct actcactact gcacaggtcg 9420 ccaaagacat agtagcactg ctcagaaaag gtgatcttgt tcacggtgtg cctcaggaaa 9480 ccgtgcttca gcatactgct ggcatacttt cttgcctccc ttcgctcctt gaagccctcc 9540 acgtgtgtgt acagccagtc caccacatcc gcccctggcc acaggtccat caaagtcagg 9600 gtagctgagc cctgggaagc tacgccagaa tgaggaacag acggggccct tcccacacag 9660 ccagggactc accaatgaca gcattggcaa tggtgatctt aagccacatg cggtcccgga 9720 tctccagtcc tgagtctggc aactgcatga cgcggacaat ggcactcatg tcactcttca 9780 cagtcagcgg tgcctcctca agctctgcag agcacacttc cctgagccca ggctcacagc 9840 gtgaacctcc atggggttga gagcaggggc cagggtcaaa cctcttatct cc~atccttg 9900 ggagatgccc ctcatcgaaa cttgagctaa gaccgggaga ttcttccccg tcccacagtg 9960 caagtccacg taggcaaggc agcccccctc ccctccccgg agagaacaag ctgttagcta 10020 tgttaggtag cagaaaagca aagcagaggc tgccatgtcc tcccaattcc cccctccgca 10080 caggcctggc aggaccctca attcatgcag atgaccagta tggccaggcc tggagggata 10140 tgtacatgta tctttgtgta cacatttgtg aaggtgttgg aagcaaacaa aaccttcata 10200 tgtaatgggc ccctgtaata gctctgatga gcaccaaagc tcaaagctag aactgaccat 10260 tgtccttcaa cctcagtttc cttgggtggg ggggggtcct gtgagctgcc acttacgtgg 10320 ggcgccaggc actgagctgg ttagtgagga agagctggtg cgtgtgatgg cgctggagca 10380 gggactcgta ccatagcggg gcagggcacc cgtcagtgct gctgtgtggg acagccaggc 10440 agccgggtcg atgggtcgca ctgggtcagc tgcatagttt ccacagcaac ggattacagg 10500 tggtaagtag gggggcagca cagaggcaga caagaaagac ccccagactg aacacagaaa 10560 ccccacccta ccccaccttt ccatggggta actcacccct tgggatggtg aagtagctcc 10620 gaggggttgg gtcccagcac ttggccactg tgagactgat gggcctacag agttgagcag 10680 accatgttgt aagtgaggcc cgcacagccc ctcccatcct gtgccactcc cacccccact 10740 tggctcccac ctcaccctgt ctgggacacg atctcccgaa gcacccgtac agcgtcgtca 10800 ttgctcatgt tctcaaagtt gacatcgttc acctacgggg tttgtggggt caggggttgg 10860 tggtgggatg tgggtgcctc ttgtccccac agtccccaca tggctcccac ctgcagcaac 10920 atgtcgcccg gctcaatgcg gccatcagca gccacggccc cgcccttcat gatggatcca 10980 atgtagatgc cgccatcacc ccggtcgttg ctctggccca cgatgctgat gcccaggaag 11040 tggtgcctct ctgcaggagg ggccgtgagc aggcccccaa agctcccgag gctgtaccca 11100 cccccagcag gcacccacag cccacaaggc ctcacccatg ttgagagtga cggtgatgat 11160 gttcagggac atggtggagt ctgtgatgct gctgaaggag gatgcctgcg gagggaccca 11220 gtgaggggct gtgtgggcac cattcagagc agacacccca cccacctgct gcctacccgg 11280 tctgtctgcc tcaagcgctg cttccgacga cggcatttgt gcttccgaac tagccgagag 11340 gaggtgctct gctctgtgga gctgctcagc ctgaggcagg agtcagaaaa gcacaaacat 11400 gtataaccag ctcggacgct caactacaaa tctccagcac gtactgacat gtgcacacgt 11460 cacccaccgg ctcgtattgt cctcctcatc tgagtcaata aagctgctag attcaagctc 11520 actgctcagt acagtggatg cactgtctgg aggtagtccc aggtcccgcc gccgatcccc 11580 tctcgggtgc ccattggtcc gggcagctgt ggggacagta gggtgggtac gactgtggga 11640 cttcagtcct aacagaatgc gggtggcctg tgcatttcaa agtttatgca gtaactctgg 11700 ggccacaggg gctaggagta ccaggctggg acctctaccc aaggatcact gcttggaaga 11760 atatgtggaa tacttccagg cttggagtat accaaaggga taccaaggg 1180:
<210> 3 <211> 858 <212> PRT
<213> Mouse <400> 3 Met Pro Ala Leu Ala Ile Met Gly Leu Ser Leu Ala Ala Phe Leu Glu Leu Gly Met Gly Ala Ser Leu Cys Leu Ser Gln Gln Phe Lys Ala Gln Gly Asp Tyr Ile Leu Gly Gly Leu Phe Pro Leu Gly Ser Thr Glu Glu Ala Thr Leu Asn Gln Arg Thr Gln Pro Asn Ser Ile Pro Cys Asn Arg Phe Ser Pro Leu Gly Leu Phe Leu Ala Met Ala Met Lys Met Ala Val Glu Glu Ile Asn Asn Gly Ser Ala Leu Leu Pro Gly Leu Arg Leu Gly Tyr Asp Leu Phe Asp Thr Cys Ser Glu Pro Val Val Thr Met Lys Ser Ser Leu Met Phe Leu Ala Lys Val Gly Ser Gln Ser Ile Ala Ala Tyr Cys Asn Tyr Thr Gln Tyr Gln Pro Arg Val Leu Ala Val Ile Gly Pro His Ser Ser Glu Leu Ala Leu Ile Thr Gly Lys Phe Phe Ser Phe Phe Leu Met Pro Gln Val Ser Tyr Ser Ala Ser Met Asp Arg Leu Ser Asp Arg Glu Thr Phe Pro Ser Phe Phe Arg Thr Val Pro Ser Asp Arg Val Gln Leu Gln Ala Val Val Thr Leu Leu Gln Asn Phe Ser Trp Asn Trp Val Ala Ala Leu Gly Ser Asp Asp Asp Tyr Gly Arg Glu Gly Leu Ser Ile Phe Ser Ser Leu Ala Asn Ala Arg Gly Ile Cys Ile Ala His Glu Gly Leu Val Pro Gln His Asp Thr Ser Gly Gln Gln Leu Gly Lys Val Leu Asp Val Leu Arg Gln Val Asn Gln Ser Lys Val Gln Val Val Val Leu Phe Ala Ser Ala Arg Ala Val Tyr Ser Leu Phe Ser Tyr Ser Ile His His Gly Leu Ser Pro Lys Val Trp Val Ala Ser Glu Ser Trp Leu Thr Ser Asp Leu Val Met Thr Leu Pro Asn Ile Ala Arg Val Gly Thr Val Leu Gly Phe Leu Gln Arg Gly Ala Leu Leu Pro Glu Phe Ser His Tyr Val Glu Thr His Leu Ala Leu Ala Ala Asp Pro A1a Phe Cys Ala Ser Leu Asn Ala Glu Leu Asp Leu Glu Glu His Val Met Gly Gln Arg Cys Pro Arg Cys Asp Asp Ile Met Leu Gln Asn Leu Ser Ser Gly Leu Leu Gln Asn Leu Ser Ala Gly Gln Leu His His Gln Ile Phe Ala Thr Tyr Ala Ala Val Tyr Ser Val Ala Gln Ala Leu His Asn Thr Leu Gln Cys Asn Val Ser His Cys His Val Ser Glu His Val Leu Pro Trp Gln Leu Leu Glu Asn Met Tyr Asn Met Ser Phe His Ala Arg Asp Leu Thr Leu Gln Phe Asp Ala Glu Gly Asn Val Asp Met Glu Tyr Asp Leu Lys Met Trp Val Trp Gln Ser Pro Thr Pro Val Leu His Thr Val Gly Thr Phe Asn Gly Thr Leu Gln Leu Ghn Gln Ser Lys Met Tyr Trp Pro Gly Asn Gln Val Pro Val Ser Gln Cys Ser Arg Gln Cys Lys Asp Gly Gln Val Arg Arg Val Lys Gly Phe His Ser Cys Cys Tyr Asp Cys Val Asp Cys Lys Ala Gly Ser Tyr Arg Lys His Pro Asp Asp Phe Thr Cys Thr Pro Cys Asn Gln Asp Gln Trp Ser Pro Glu Lys Ser Thr Ala Cys Leu Pro Arg Arg Pro Lys Phe Leu Ala Trp Gly Glu Pro Val Val Leu Ser Leu Leu Leu Leu Leu Cys Leu Val Leu Gly Leu Ala Leu Ala Ala Leu Gly Leu Ser Val His His Trp Asp Ser Pro Leu Val Gln Ala Ser Gly Gly Ser Gln Phe Cys Phe Gly Leu Ile Cys Leu Gly Leu Phe Cys Leu Ser Val Leu Leu Phe Pro Gly Arg Pro Ser Ser Ala Ser Cys Leu Ala Gln Gln Pro Met Ala His Leu Pro Leu Thr Gly Cys Leu Ser Thr Leu Phe Leu Gln Ala Ala Glu Thr Phe Val Glu Ser Glu Leu Pro Leu Ser Trp Ala Asn Trp Leu Cys Ser Tyr Leu Arg Gly Leu Trp Ala Trp Leu Val Val Leu Leu Ala Thr Phe Val Glu Ala Ala Leu Cys Ala Trp Tyr Leu Ile Ala Phe Pro Pro Glu Val Val Thr Asp Trp Ser Val Leu Pro Thr Glu Val Leu Glu His Cys His Val Arg Ser Trp Val Ser Leu Gly Leu Val His Ile Thr Asn Ala Met Leu Ala Phe Leu Cys Phe Leu Gly Thr Phe Leu Val Gln Ser Gln Pro Gly Arg Tyr Asn Arg Ala Arg Gly Leu Thr Phe Ala Met Leu Ala Tyr Phe Ile Thr Trp Val Ser Phe Val Pro Leu Leu Ala Asn Val Gln Val Ala Tyr Gln Pro A1a Val Gln Met Gly Ala Ile Leu Val Cys Ala Leu Gly Ile Leu Val Thr Phe His Leu Pro Lys Cys Tyr Val Leu Leu Trp Leu Pro Lys Leu Asn Thr Gln Glu Phe Phe Leu Gly Arg Asn Ala Lys Lys Ala Ala Asp Glu Asn Ser Gly Gly Gly Glu Ala Ala Gln Gly His Asn Glu <210> 4 <211> 2559 <212> DNA
<213> Homo Sapiens <400> 4 atgctgggcc ctgctgtcct gggcctcagc ctctgggctc tcctgcaccc tgggacgggg 60 gccccattgt gcctgtcaca gcaacttagg atgaaggggg actacgtgct gggggggctg 120 ttccccctgg gcgaggccga ggaggctggc ctccgcagcc ggacacggcc cagcagccct 180 gtgtgcacca ggttctcctc aaacggcctg ctctgggcac tggccatgaa aatggccgtg 240 gaggagatca acaacaagtc ggatctgctg cccgggctgc gcctgggcta cgacctcttt 300 gatacgtgct cggagcctgt ggtggccatg aagcccagcc tcatgttcct ggccaaggca 360 ggcagccgcg acatcgccgc ctactgcaac tacacgcagt accagccccg tgtgctggct 420 gtcatcgggc cccactcgtc agagctcgcc atggtcaccg gcaagttctt cagcttcttc 480 ctcatgcccc aggtcagcta cggtgctagc atggagctgc tgagcgcccg ggagaccttc 540 ccctccttct tccgcaccgt gcccagcgac cgtgtgcagc tgacggccgc cgcggagctg 600 ctgcaggagt tcggctggaa ctgggtggcc gccctgggca gcgacgacga gtacggccgg 660 cagggcctga gcatcttctc ggccctggcc tcggcacgcg gcatctgcat cgcgcacgag 720 ggcctggtgc cgctgccccg tgccgatgac tcgcggctgg ggaaggtgca ggacgtcctg 780 caccaggtga accagagcag cgtgcaggtg gtgctgctgt tcgcctccgt gcacgccgcc 840 cacgccctct tcaactacag catcagcagc aggctctcgc ccaaggtgtg ggtggccagc 900 gaggcctggc tgacctctga cctggtcatg gggctgcccg gcatggccca gatgggcacg 960 gtgcttggct tcctccagag gggtgcccag ctgcacgagt tcccccagta cgtgaagacg 1020 cacctggccc tggccaccga cccggccttc tgctctgccc tgggcgagag ggagcagggt 1080 ctggaggagg acgtggtggg ccagcgctgc ccgcagtgtg actgcatcac gctgcagaac 1140 gtgagcgcag ggctaaatca ccaccagacg ttctctgtct acgcagctgt gtatagcgtg 1200 gcccaggccc tgcacaacac tcttcagtgc aacgcctcag gctgccccgc gcaggacccc 1260 gtgaagccct ggcagctcct ggagaacatg tacaacctga ccttccacgt gggcgggctg 1320 ccgctgcggt tcgacagcag cggaaacgtg gacatggagt acgacctgaa gctgtgggtg 1380 tggcagggct cagtgcccag gctccacgac gtgggcaggt tcaacggcag cctcaggaca 1440 gagcgcctga agatccgctg gcacacgtct gacaaccaga agcccgtgtc ccggtgctcg 1500 cggcagtgcc aggagggcca ggtgcgccgg gtcaaggggt tccactcctg ctgctacgac 1560 tgtgtggact gcgaggcggg cagctaccgg caaaacccag acgacatcgc ctgcaccttt 1620 tgtggccagg atgagtggtc cccggagcga agcacacgct gcttccgccg caggtctcgg 1680 ttcctggcat ggggcgagcc ggctgtgctg ctgctgctcc tgctgctgag cctggcgctg 1740 ggccttgtgc tggctgcttt ggggctgttc gttcaccatc gggacagccc actggttcag 1800 gcctcggggg ggcccctggc ctgctttggc ctggtgtgcc tgggcctggt ctgcctcagc 1860 gtcctcctgt tccctggcca gcccagccct gcccgatgcc tggcccagca gcccttgtcc 1920 cacctcccgc tcacgggctg cctgagcaca ctcttcctgc aggcggccga gatcttcgtg 1980 gagtcagaac tgcctctgag ctgggcagac cggctgagtg gctgcctgcg ggggccctgg 2040 gcctggctgg tggtgctgct ggccatgctg gtggaggtcg cactgtgcac ctggtacctg 2100 gtggccttcc cgccggaggt ggtgacggac tggcacatgc tgcccacgga ggcgctggtg 2160 cactgccgca cacgctcctg ggtcagcttc ggcctagcgc acgccaccaa tgccacgctg 2220 gcctttctct gcttcctggg cactttcctg gtgcggagcc agccgggccg ctacaaccgt 2280 gcccgtggcc tcacctttgc catgctggcc tacttcatca cctgggtctc ctttgtgccc 2340 ctcctggcca atgtgcaggt ggtcctcagg cccgccgtgc agatgggcgc cctcctgctc 2400 tgtgtcctgg gcatcctggc tgccttccac ctgcccaggt gttacctgct catgcggcag 2460 ccagggctca acacccccga gttcttcctg ggagggggcc ctggggatgc ccaaggccag 2520 aatgacggga acacaggaaa tcaggggaaa catgagtga 2559 <210> 5 <211> 852 <212> PRT
<213> Homo sapiens <400> 5 Met Leu Gly Pro Ala Val Leu Gly Leu Ser Leu Trp Ala Leu Leu His Pro Gly Thr Gly Ala Pro Leu Cys Leu Ser Gln Gln Leu Arg Met Lys Gly Asp Tyr Va1 Leu Gly Gly Leu Phe Pro Leu Gly Glu Ala Glu Glu Ala Gly Leu Arg Ser Arg Thr Arg Pro Ser Ser Pro Val Cys Thr Arg Phe Ser Ser Asn Gly Leu Leu Trp Ala Leu Ala Met Lys Met Ala Val Glu Glu Ile Asn Asn Lys Ser Asp Leu Leu Pro Gly Leu Arg Leu Gly Tyr Asp Leu Phe Asp Thr Cys Ser Glu Pro Val Val Ala Met Lys Pro Ser Leu Met Phe Leu Ala Lys Ala Gly Ser Arg Asp Ile Ala Ala Tyr Cys Asn Tyr Thr Gln Tyr Gln Pro Arg Val Leu Ala Val Ile Gly Pro His Ser Ser Glu Leu Ala Met Val Thr Gly Lys Phe Phe Ser Phe Phe Leu Met Pro Gln Val Ser Tyr Gly Ala Ser Met Glu Leu Leu Ser Ala Arg Glu Thr Phe Pro Ser Phe Phe Arg Thr Val Pro Ser~Asp Arg Val Gln Leu Thr Ala Ala Ala Glu Leu Leu Gln Glu Phe Gly Trp Asn Trp Val Ala Ala Leu Gly Ser Asp Asp Glu Tyr Gly Arg Gln Gly Leu Ser Ile Phe Ser Ala Leu Ala Ser Ala Arg Gly Ile Cys Ile Ala His Glu Gly Leu Va1 Pro Leu Pro Arg Ala Asp Asp Ser Arg Leu Gly Lys Val Gln Asp Val Leu His Gln Val Asn Gln Ser Ser Val Gln Val Val Leu Leu Phe Ala Ser Val His Ala Ala His Ala Leu Phe Asn Tyr Ser Ile Ser Ser Arg Leu Ser Pro Lys Val Trp Val Ala Ser Glu Ala Trp Leu Thr Ser Asp Leu Val Met Gly Leu Pro Gly Met Ala Gln Met Gly Thr Val Leu Gly Phe Leu Gln Arg Gly Ala Gln Leu His Glu Phe Pro Gln Tyr Val Lys Thr His Leu Ala Leu Ala Thr Asp Pro Ala Phe Cys Ser Ala Leu Gly Glu Arg Glu Gln Gly Leu Glu G1u Asp Val Val Gly Gln Arg Cys Pro Gln Cys Asp Cys Ile Thr Leu G1n Asn Val Ser Ala Gly Leu Asn His His Gln Thr Phe Ser Val Tyr Ala Ala Val Tyr Ser Val Ala Gln Ala Leu His Asn Thr Leu Gln Cys Asn Ala Ser Gly Cys Pro Ala Gln Asp Pro Val Lys Pro Trp Gln Leu Leu Glu Asn Met Tyr Asn Leu Thr Phe His Val Gly Gly Leu Pro Leu Arg Phe Asp Ser Ser Gly Asn Val Asp Met Glu Tyr Asp Leu Lys Leu Trp Val Trp Gln Gly Ser Val Pro Arg Leu His Asp Val Gly Arg Phe Asn Gly Ser Leu Arg Thr Glu Arg Leu Lys Ile Arg Trp His Thr Ser Asp Asn Gln Lys Pro Val Ser Arg Cys Ser Arg Gln Cys Gln Glu Gly Gln Val Arg Arg Val Lys Gly Phe His Ser Cys Cys Tyr Asp Cys Val Asp Cys Glu Ala Gly Ser Tyr Arg Gln Asn Pro Asp Asp Ile Ala Cys Thr Phe Cys Gly Gln Asp Glu Trp Ser Pro Glu Arg Ser Thr Arg Cys Phe Arg Arg Arg Ser Arg ., Phe Leu Ala Trp Gly Glu Pro Ala Val Leu Leu Leu Leu Leu Leu Leu Ser Leu Ala Leu Gly Leu Val Leu Ala Ala Leu Gly Leu Phe Val His His Arg Asp Ser Pro Leu Val Gln Ala Ser Gly Gly Pro Leu Ala Cys Phe Gly Leu Val Cys Leu Gly Leu Val Cys Leu Ser Val Leu Leu Phe Pro Gly Gln Pro Ser Pro Ala Arg Cys Leu Ala Gln Gln Pro Leu Ser His Leu Pro Leu Thr Gly Cys Leu Ser Thr Leu Phe Leu Gln Ala Ala Glu Ile Phe Val Glu Ser Glu Leu Pro Leu Ser Trp Ala Asp Arg Leu.

Ser Gly Cys Leu Arg Gly Pro Trp Ala Trp Leu Val Val Leu Leu Ala Met Leu Val Glu Val Ala Leu Cys Thr Trp Tyr Leu Val Ala Phe Pro Pro Glu Val Val Thr Asp Trp His Met Leu Pro Thr Glu Ala Leu Val His Cys Arg Thr Arg Ser Trp Val Ser Phe Gly Leu Ala His Ala Thr 725 730 ' 735 Asn Ala Thr Leu Ala Phe Leu Cys Phe Leu Gly Thr Phe Leu Val Arg Ser Gln Pro Gly Arg Tyr Asn Arg Ala Arg Gly Leu Thr Phe Ala Met 755 ~ 760 765 Leu Ala Tyr Phe Ile Thr Trp Val Ser Phe Val Pro Leu Leu Ala Asn Val Gln Val Val Leu Arg Pro Ala Val Gln Met Gly Ala Leu Leu Leu Cys Val Leu Gly Ile Leu Ala Ala Phe His Leu Pro Arg Cys Tyr Leu Leu Met Arg Gln Pro Gly Leu Asn Thr Pro Glu Phe Phe Leu Gly Gly Gly Pro Gly Asp Ala Gln Gly Gln Asn Asp Gly Asn Thr Gly Asn Gln Gly Lys His Glu <210> 6 <211> 20 <212> DNA
<213> Mouse <400> 6 cactagagct gccaccttcc 20 <210> 7 <211> 20 <212> DNA
<213> Mouse <400> 7 ccctcagcac cactttttgt 20 <210> 8 <211> 20 <212> DNA
<213> Mouse <400> 8 acaaaaagtg gtgctgaggg 20 <210> 9 <211> 20 <212> DNA
<213> Mouse <400> 9 caggagaccc aaaggatcaa 20 <210> 10 <211> 20 <212> DNA
<213> Mouse <400> 10 gcttcagaaa atcgaggcac 20 <210> 11 <211> 20 <212> DNA
<213> Mouse <400> 11 gcatgggcta tgataggtgg 20 <210> 12 <211> 16 <212> DNA
<213> Mouse <400> 12 tgttgatccc acagcg 16 <210> 13 <211> 20 <212> DNA
<213> Mouse , <400> 13 caggaaatgt ccacttctgc 20 <210> 14 <211> 18 <212> DNA
<213> Mouse <400> 14 tctatcttgc atccagcc 18 <210> 15 <211> 16 <212> DNA
<213> Mouse <400> 15 gtgctgtgac tgtgcg 16 <210> 16 <211> 18 <212> DNA
<213> Mouse <400> 16 cgcagcattt atttggag 18 <210> 17 <211> 19 <212> DNA
<213> Mouse <400> 17 ccgacccttt aggagacac 19 <210> 18 <211> 20 <212> DNA
<213> Mouse <400> 18 tgtgacttcc tcttccccac 20 <210> 19 <211> 20 <212> DNA
<213> Mouse <400> 19 tgagccactc cagatgtcag 20 <210>20 <211>20 <212>DNA

<213>Mouse <400> 20 .
ccaacgtgca gtcaagaaaa 20 <210> 21 <211> 20 <212> DNA
<213> Mouse <400> 21 ccaacgtgca gtcaagaaaa 20 <210> 22 <211> 20 <212> DNA
<213> Mouse <400> 22 cgagagacaa agtggtgctg 20 <210> 23 <211> 20 <212> DNA
<213> Mouse <400> 23 ttatgaaggc cctcaccaac 20 <210> 24 <211> 20 <212> DNA
<213> Mouse <400> 24 ccagctccta gaattgcctg 20 <210> 25 <211> 20 <212> DNA
<213> Mouse <400> 25 gcagtctccc gaaacaagtc 20 <210> 26 <211> 20 <212> DNA
<213> Mouse <400> 26 atagaggaat gggtgcgatg 20 <210> 27 <211> 20 <212> DNA
<213> Mouse <400> 27 taccaggagg ggtcagtcag 20 <210> 28 <211> 20 <212> DNA
<213> Mouse <400> 28 tacaagcgag ctgaccaatg 20 <210> 29 <211> 20 <212> DNA
<213> Mouse <400> 29 ccaatcagct cgagttagcc 20 <210> 30 <211> 20 <212> DNA
<213> Mouse <400> 30 tgccattgtg gatgttcact 20 <210> 31 <211> 20 <212> DNA
<213> Mouse <400> 31 gagtccgagg tcggtcaata 20 <210> 32 <211> 20 <212> DNA
<213> Mouse <400> 32 gctggcttct gtaggtcagg 20 <210> 33 <211> 20 <212> DNA
<213> Mouse <400> 33 tatgagggtc aagggtcagg 20 <210> 34 <211> 20 <212> DNA
<213> Mouse <400> 34 cgctttggtg agaactagcc 20 <210> 35 <211> 20 <212> DNA
<213> Mouse <400> 35 catgtggagt tgtgggagtg ' 20 <210> 36 <211> 20 <212> DNA
<213> Mouse <400> 36 aatgggcaga agacagatgg 20 <210> 37 <211> 20 <212> DNA
<213> Mouse <400> 37 tatcagggtc tgtgaagccc 20 <210> 38 <211> 20 <212> DNA
<213> Mouse <400> 38 atacaggacc ctttaccccg 20 <210> 39 <211> 20 <212> DNA
<213> Mouse <400> 39 cagtgtttct aggtccccca 20 <210> 40 <211> 20 <212> DNA
<213> Mouse <400> 40 gcctctgtct gccatctctc 20 <210> 41 <211> 20 <212> DNA
<213> Mouse <400> 41 ataatgttac ctgcaggcgg 20 <210> 42 <211> 20 <212> DNA
<213> Mouse <400> 42 ctggaaacac ccatgtcctc 20 <210> 43 <211> 20 <212> DNA
<213> Mouse <400> 43 cgggcacatg gacactttta 20 <210> 44 <211> 20 <212> DNA
<213> Mouse <400> 44 gagcatgaag tgcaaggtga 20 <210> 45 <211> 20 <212> DNA
<213> Mouse <400> 45 cgtaggtggc acagttgaga 20 <210> 46 <211> 20 <212> DNA
<213> Mouse <400> 46 gctgttagtg aggtc,agggc 20 <210> 47 <211> 20 <212> DNA
<213> Mouse <400> 47 cgtaggtggc acagttgaga 20 <210> 48 <211> 20 <212> DNA
<213> Mouse <400> 48 gagcatgaag tgcaaggtga 20 <210> 49 <211> 20 <212> DNA
<213> Mouse <400> 49 tcattttcct agcctcggtg 20 <210> 50 <211> 22 <212> DNA
<213> Mouse <400> 50 tctaagaaga tgatgcagac cc 22 <210> 51 <211> 20 <212> DNA
<213> Mouse <400> 51 tgtccttcag ggatagtgcc 20 <210> 52 <211> 20 <212> DNA
<213> Mouse <400> 52 ggcttcagcc tcaagttctg 20 <210> 53 <211> 20 <212> DNA
<213> Mouse <400> 53 aaaacaacca agttgccctg 20 <210> 54 <211> 20 <212> DNA
<213> Mouse <400> 54 ggcactgaaa tgacctggat 20 <210> 55 <211> 20 <212> DNA
<213> Mouse <400> 55 aacaattcaa gcaacctcgg 20 <210> 56 <211> 20 <212> DNA
<213> Mouse <400> 56 ctgttccttc ccagactcca 20 <210> 57 <211> 20 <212> DNA
<213> Mouse <400> 57 ttcagtcacg caaacctgag . 20 <210> 58 <211> 20 <212> DNA
<213> Mouse <400> 58 gcccaggact ttgtcactgt 20 <210> 59 <211> 20 <212> DNA
<213> Mouse <400> 59 ggtaacctgc agctccactc 20 <210> 60 <211> 20 <212> DNA
<213> Mouse , <400> 60 gggacatgct cttggttcat 20 <210> 61 <211> 20 <212> DNA
<213> Mouse <400> 61 gaacaaagcc gggtgattta 20 <210> 62 <211> 20 <212> DNA
<213> Mouse <400> 62 gccctcagtt ctcctagcct 20 <210> 63 <211> 20 <212> DNA
<213> Mouse <400> 63 ggcagagaag actggtggag 20 <210> 64 <211> 20 <212> DNA
<213> Mouse <400> 64 cccagactta gcgtctcagg 20 <210> 65 <211> 20 <212> DNA
<213> Mouse <400> 65 agcagagacc tttggactcg 20 <210> 66 <211> 20 <212> DNA
<213> Mouse <400> 66 gaaggctgag tgagtcccag 20 <210> 67 <211> 20 <212> DNA
<213> Mouse <400> 67 ttgcacgagg agaaggtttt 20 <210> 68 <211> 20 <212> DNA
<213> Mouse <400> 68 gatgccaacg agacctgaat 20 <210> 69 <211> 20 <212> DNA
<213> Mouse <400> 69 agaagccaaa accctcacct 20 <210> 70 <211> 20 <212> DNA
<213> Mouse <400> 70 aaaaagccct gcaagaactt 20 <210> 71 <211> 20 <212> DNA
<213> Mouse <400> 71 attcaggtct cgttggcatc 20 <210> 72 <211> 20 <212> DNA
<213> Mouse <400> 72 tgtccgcagt gtggaaacta 20 <210> 73 <211> 20 <212> DNA
<213> Mouse <400> 73 atgtccaggg tagagagccc 20 <210> 74 <211> 20 <212> DNA
<213> Mouse <400> 74 ggagttctcc taccctggct 20 <210> 75 <211> 20 <212> DNA
<213> Mouse <400> 75 gaggctctga gcagtgtcaa 20 <210> 76 <211> 14 <212> DNA
<213> Mouse <400> 76 gcgatgttgt tgcg 14 <210> 77 <211> 18 <212> DNA
<213> Mouse <400> 77 cagtgtcttt ccacattt 18 <210> 78 <211> 27 <212> DNA
<213> Mouse <400> 78 aggcatattg tataataaat ttgtagt 27 <210> 79 <211> 19 <212> DNA
<213> Mouse <400> 79 ccggatgact ctacttgac 19 <210> 80 <211> 20 <212> DNA
<213> Mouse <400> 80 gctgtttatg gggtcgagaa 20 <210> 81 <211> 20 <212> DNA
<213> Mouse <400> 81 aatttctgaa gcagggggat 20 <210> 82 <211> 20 <212> DNA
<213> Mouse <400> 82 tccccctgct tcagaaatta 20 <210> 83 <211> 20 <212> DNA
<213> Mouse <400> 83 agggggatga ttgtgagtga 20 <210> 84 <211> 27 <212> DNA
<213> Mouse <400> 84 cttctttaat caatctctgt ctctgtg 27 <210> 85 <211> 20 <212> DNA
<213> Mouse <400> 85 gggcacatat gaacctcctg 20 <210> 86 <211> 20 <212> DNA
<213> Mouse <400> 86 ccaaactctt agcttcttca 20 <210> 87 <211> 21 <212> DNA
<213> Mouse <400> 87 acacagaaga cactgaagaa c 21 <210> 88 <211> 22 <212> DNA
<213> Mouse <400> 88 cagttgttag aagcaggatc cc 22 <210> 89 <211> 23 <212> DNA
<213> Mouse <400> 89 aggtgcatat acctgggata ctc 23 <210> 90 <211> 21 <212> DNA
<213> Mouse <400> 90 agagtttggt ctcttcccct g 21 <210> 91 <211> 23 <212> DNA
<213> Mouse <400> 91 tatccaacac atttatgtct gcg 23 <210> 92 <211> 20 <212> DNA
<213> Mouse <400> 92 gccagtgtgc tgaaagactg 20 <210> 93 <211> 20 <212> DNA
<213> Mouse <400> 93 agggacctgg agacatcctt 20 <210> 94 <211> 23 <212> DNA
<213> Mouse <400> 94 ctgtaggctg cttttatctt ttg 23 <210> 95 <211> 20 <212> DNA
<213> Mouse <400> 95 tgccccttca gcacatgcca 20 <210> 96 <211> 23 <212> DNA
<213> Mouse <400> 96 tgcagtgtga catgtgcata gat 23 <210> 97 <211> 21 <212> DNA
<213> Mouse <400> 97 ggaaagccag gctacgcaga a 21 <210> 98 <211> 23 <212> DNA
<213> Mouse <400> 98 ctgtaggctg cttttatctt ttg 23 <210> 99 <211> 20 <212> DNA
<213> Mouse <400> 99 tgccccttca gcacatgcca 20 <210> 100 <211> 22 <212> DNA
<213> Mouse <400> 100 tagtgtggtt cctgactaac ct 22 <210> 101 <211> 22 <212> DNA
<213> Mouse <400> 101 cggtctacat agtgagtgat tc 22 <210> 102 <211> 22 <212> DNA
<213> Mouse <400> 102 aaaagcatcc tgcatccttc tg 22 <210> 103 <211> 22 <212> DNA
<213> Mouse <400> 103 gggttataca gagaaaccct gt 22 <210> 104 <211> 20 <212> DNA
<213> Mouse <400> 104 ttccaagctc acacatcagc 20 <210> 105 <211> 20 <212> DNA
<213> Mouse <400> 105 gtgctgctct gcattgagtg 20 <210> 106 <211> 20 <212> DNA
<213> Mouse <400> 106 gacagtgtgg gagaatccgt 20 <210> 107 <211> 20 <212> DNA
<213> Mouse <400> 107 cccaaggcat aggtcacaat 20 <210> 108 <211> 20 <212> DNA
<213> Mouse <400> 108 attgtgacct atgccttggg 20 <210> 109 <211> 20 <212> DNA
<213> Mouse <400> 109 cgaaggaccg tcatctgagt 20 <210> 110 <211> 20 <212> DNA
<213> Mouse <400> 110 ggctttgatg tgaaaaaggc 20 <210> 111 <211> 20 <212> DNA
<213> Mouse <400> 111 agctcctcat cgctcatgtt 20 <210> 112 <211> 20 <212> DNA
<213> Mouse <400> 112 tggaacatct ctgtcggaag 20 <210> 113 <211> 20 <212> DNA
<213> Mouse <400> 113 ggctctcatt gccaccttta 20 <210> 114 <211> 20 <212> DNA
<213> Mouse <400> 114 ccagagaaca ggagacctgc 20 <210> 115 <211> 20 <212> DNA
<213> Mouse <400> 115 gtgctggata cactggcaga 20 <210> 116 <211> 20 <212> DNA
<213> Mouse <400> 116 gcgagacgag tgggtagttc 20 <210> 117 <211> 20 <212> DNA
<213> Mouse <400> 117 acactgaaac ctcgcttgct 20 <210> 118 <211> 20 <212> DNA
<213> Mouse <400> 118 agcaagcgag gtttcagtgt 20 <210> 119 <211> 20 <212> DNA
<213> Mouse <400> 119 acggggcttg atccttttat 20 <210> 120 <211> 25 <212> DNA
<213> Mouse <400> 120 aagttcatgg gcctcaccac ctgtc 25 <210> 121 <211> 22 <212> DNA
<213> Mouse <400> 121 tactagctac ccttcacata cc 22 <210> 122 <211> 21 <212> DNA
<213> Mouse <400> 122 acctagccac tgtctcagtc t 21 <210> 123 <211> 21 <212> DNA
<213> Mouse <400> 123 acagaagcag catttacaca g 21 <210> 124 <211> 20 <212> DNA
<213> Mouse <400> 124 tgggacagct tcctcaagat r 20 <210> 125 <211> 20 <212> DNA
<213> Mouse <400> 125 aatgggaatt gtgctcttgg 20 <210> 126 <211> 20 <212> DNA
<213> Mouse <400> 126 gggcatctgg caaagattta 20 <210> 127 <211> 20 <212> DNA
<213> Mouse <400> 127 agataacctg tgtgtcccgc 20 <210> 128 <211> 20 <212> DNA
<213> Mouse <400> 128 gatgtccgag aagggatgtg 20 <210> 129 <211> 20 <212> DNA
<213> Mouse <400> 129 tgtcagcttt gagtgcatcc 20 <210> 130 <211> 20 <212> DNA
<213> Mouse <400> 130 acatgcaggc tgtttgacct 20 <210> 131 <211> 20 <212> DNA
<213> Mouse <400> 131 tgtcagcttt gagtgcatcc 20 <210> 132 <211> 20 <212> DNA
<213> Mouse <400> 132 gtgctctgca gacaaaccaa 20 <210> 133 <211> 20 <212> DNA
<213> Mouse <400> 133 gagccatttt gacccttaaa 20 <210> 134 <211> 20 <212> DNA
<213> Mouse <400> 134 tttcagggtc aaaatggctc 20 <210> 135 <211> 17 <212> DNA
<213> Mouse <400> 135 tcgacagcaa ctgtgcg 17 <210> 136 <211> 20 <212> DNA
<213> Mouse <400> 136 ggtgagagtg gggagatgaa 20 <210> 137 <211> 20 <212> DNA
<213> Mouse <400> 137 cccgggtgag tttaagaacc 20 <210> 138 <211> 20 <212> DNA
<213> Mouse <400> 138 ggtgagagtg gggagatgaa 20 <210> 139 <211> 20 <212> DNA
<213> Mouse <400> 139 aggttaggcc caatttcctg 20 <210> 140 <211> 20 <212> DNA
<213> Mouse <400> 140 ccagggttgc tgtactgaga 20 <210> 141 <211> 20 <212> DNA
<213> Mouse <400> 141 caggttaggc ccaatttcct 20 <210> 142 <211> 20 <212> DNA
<213> Mouse <400> 142 ggtcagagtc cttccttccc 20 <210> 143 <211> 20 <212> DNA
<213> Mouse <400> 143 tccaacttca caggaaaccc 20 <210> 144 <211> 20 <212> DNA
<213> Mouse <400> 144 tttcctgtga agttggaggg 20 <210> 145 <211> 20 <212> DNA
<213> Mouse <400> 145 cacccatatg gcaaacatca .20 <210> 146 <211> 20 <212> DNA
<213> Mouse <400> 146 ggtcagagtc cttccttccc 20 <210> 147 <211> 20 <212> DNA
<213> Mouse <400> 147 tccaacttca.caggaaaccc 20 <210> 148 <211> 20 <212> DNA
<213> Mouse <400> 148 tgatgtttgc catatgggtg 20 <210> 149 <211> 20 <212> DNA
<213> Mouse <400> 149 gcttgctgct tccgatatgt 20 <210> 150 <211> 19 <212> DNA
<213> Mouse <400> 150 ggaaaaggga gtcgccata 19 <210> 151 <211> 20 <212> DNA
<213> Mouse <400> 151 gagccgccta actctcacac 20 <210> 152 <211> 19 <212> DNA
<213> Mouse <400> 152 aggggataac ctgcatagg 19 <210> 153 <211> 20 <212> DNA
<213> Mouse <400> 153 , acaaaattgc tcatttgccc 20 <210> 154 <211> 20 <212> DNA
<213> Mouse <400> 154 ccatccccac tagccagata~ 20 <210> 155 <211> 20 <212> DNA
<213> Mouse <400> 155 gtcccctttg tcacagcaag 20 <210> 156 <211> 20 <212> DNA
<213> Mouse <400> 156 tgagcacagg atagctccac 20 <210'> 157 <211> 20 <212> DNA
<213> Mouse <400> 157 aaaagaacac ctgtttgggg 20 <210> 158 <211> 19 <212> DNA
<213> Mouse <400> 158 taaacctcgg ctgtgtgag 19 <210> 159 <211> 20 <212> DNA
<213> Mouse <400> 159 ccctcagtga cttcctgtga 20 <210> 160 <211> 20 <212> DNA
<213> Mouse <400> 160 caaaaccaca tggttaccga 20 <210> 161 <211> 20 <212> DNA
<213> Mouse <400> 161 gccctattgc caaatgactt 20 <210> 162 <211> 20 <212> DNA
<213> Mouse <400> 162 ggcagaaagg aatcagaagc 20 <210> 163 <211> 20 <212> DNA
<213> Mouse <400> 163 cacattagcc attgtcctgg 20 <210> 164 <211> 20 <212> DNA
<213> Mouse <400> 164 tcctttatgt ccaacagcca 20 <210> 165 <211> 20 <212> DNA
<213> Mouse <400> 165 catggtctgt gatgtgacca 20 <210> 166 <211> 20 <212> DNA
<213> Mouse <400> 166 atacccttgg tgagagcagg 20 <210> 167 <211> 20 <212> DNA
<213> Mouse <400> 167 gctgtcaaat gagaaaggca 20 <210> 168 <211> 20 <212> DNA
<213> Mouse <400> 168 tatttcatgc tgggaccaaa 20 <210> 169 <211> 20 <212> DNA
<213> Mouse <400> 169 agagaaaaac agtgggggtg 20 <210> 170 <211> 20 <212> DNA
<213> Mouse <400> 170 cgggtcctct cttcaccata 20 <210> 171 <211> 20 <212> DNA
<213> Mouse <400> 171 ctacatttcc ctgagctgcc 20 <210> 172 <211> 20 <212> DNA
<213> Mouse <400> 172 gttgaccatg tcggtaaccc 20 <210> 173 <211> 20 <212> DNA
<213> Mouse <400> 173 ccacctcacg gaaactgaat 20 <210> 174 <211> 20 <212> DNA
<213> Mouse <400> 174 ggtgtttggc tcacaaacct 20 <210> 175 <211> 20 <212> DNA
<213> Mouse <400> 175 gatgcacaca caaaaatccg 20 <210> 176 <211> 20 <212> DNA
<213> Mouse <400> 176 atcacccacc agaacgaaaa 20 <210> 177 <211> 20 <212> DNA
<213> Mouse <400> 177 accctccagg agtaggtgct 20 <210>178 <211>20 <212>DNA

<213>Mouse <400> 178 gatgagacag tgggcaaggt 20 <210> 179 <211> 20 <212> DNA
<213> Mouse <400> 179 ttgtcaatag caccaagcca ' 20 <210> 180 <211> 20 <212> DNA
<213> Mouse <400> 180 gccttaatag cccccttgtt 20 <210> 181 <211> 20 <212> DNA
<213> Mouse v1 <400> 181 gcactcagca ttgcacagat 20 <210> 182 <211> 20 <212> DNA
<213> Mouse <400> 182 ggacggacaa ttctggaaaa 20 <210> 183 <211> 20 <212> DNA
<213> Mouse <400> 183 ctatcacacc tccgatgcct 20 <210> 184 <211> 20 <212> DNA
<213> Mouse <400> 184 caagctggta gaatccccaa 20 <210> 185 <211> 20 <212> DNA
<213> Mouse <400> 185 tctttggaga agcagaccgt 20 <210> 186 <211> 20 <212> DNA
<213> Mouse <400> 186 tacagcatat gcatgccagg , 20 <210> 187 <211> 20 <212> DNA
<213> Mouse <400> 187 attcctcagg gcattacacg 20 <210> 188 <211> 20 <212> DNA
<213> Mouse <400> 188 gcaatctctt gtgtccaggc 20 <210> 189 <211> 20 <212> DNA
<213> Mouse <400> 189 attcctcagg gcattacacg 20 <210> 190 <211> 20 <212> DNA
<213> Mouse <400> 190 tacagcatat gcatgccagg 20 <210> 191 <211> 20 <212> DNA
<213> Mouse <400> 191 ggcctggaca caagagattg 20 <210> 192 <211> 20 <212> DNA
<213> Mouse <400> 192 aagtgggtgg acagtgaagg 20 <210> 193 <211> 20 <212> DNA
<213> Mouse <400> 193 cagcttcctc catcttctgg 20 <210> 194 <211> 20 <212> DNA
<213> Mouse <400> 194 agagcctcca gtagatggca 20 <210> 195 <211> 20 <212> DNA
<213> Mouse <400> 195 tcgtggacaa gctccttctt 20 <210> 196 <211> 20 <212> DNA
<213> Mouse <400> 196 catcgagtat gtcaatggcg 20 <210> 197 <211> 20 <212> DNA
<213> Mouse <400> 197 ttgtccagtt ttaggtcccg 20 <210> 198 <211> 20 <212> DNA
<213> Mouse <400> 198 cagactgggt tttccgacat 20 <210> 199 -<211> 20 <212> DNA
<213> Mouse <400> 199 gtcaaagttg tccaggccat 20 <210> 200 <211> 18 <212> DNA
<213> Mouse <400> 200 aggacggacc ccaagatg 18 <210> 201 <211> 20 <212> DNA
<213> Mouse <400> 201 tgtctcgcac ttcctcacag 20 <210> 202 <211> 20 <212> DNA
<213> Mouse <400> 202 ccagaagatg gaggaagctg 20 <210> 203 <211> 20 <212> DNA
<213> Mouse <400> 203 tctactggag gctcttggga 20 <210> 204 <211> 20 <212> DNA
<213> Mouse <400> 204 gaaaaacgac cagatttacg 20 <210> 205 <211> 20 <212> DNA
<213> Mouse <400> 205 gatctcagca gcatagaacc 20 <210> 206 <211> 20 <212> DNA
<213> Mouse <400> 206 acacattaag ctgacggact 20 <210> 207 <211> 20 <212> DNA
<213> Mouse <400> 207 caaacataag gacacccagt 20 <210> 208 <211> 20 <212> DNA
<213> Mouse <400> 208 actgggtgtc cttatgtttg 20 <210> 209 <211> 20 <212> DNA
<213> Mouse <400> 209 cctctctttg ggatccttat 20 <210> 210 <211> 20 <212> DNA
<213> Mouse <400> 210 gtcataaaga ggatcgacca 20 <210> 211 <211> 20 <212> DNA
<213> Mouse <400> 211 gctctgtcta gaagtgcctg 20 <210> 212 <211> 18 <212> DNA
<213> Mouse <400> 212 accaagaccg aagagggg , 18 <210>213 <211>22 <212>DNA

<213>Mouse <400> 213 ggcattacac gctaactttt cc 22 <210> 214 <211> 20 <212> DNA
<213> Mouse <400> 214 agtgccacca acctggtaag 20 <210> 215 <211> 18 <212> DNA
<213> Mouse <400> 215 aagtgcctgc agggatgc 18 <210> 216 <211> 20 <212> DNA
<213> Mouse <400> 216 tgctttggtg agcaatgttt 20 <210> 217 <211> 20 <212> DNA
<213> Mouse <400> 217 agggacaccc ttaccaggtt 20 <210> 218 <211> 20 <212> DNA
<213> Mouse <400> 218 ctgatgcttt ggtgagcaat 20 <210> 219 <211> 19 <212> DNA
<213> Mouse <400> 219 gggacaccct taccaggtt 19 <210> 220 <211> 20 <212> DNA
<213> Mouse <400> 220 acaggacaaa tgctgggttg 20 <210> 221 <211> 20 <212> DNA
<213> Mouse <400> 221 gtggtaaaga acgcttggct 20 <210> 222 <211> 24 <212> DNA
<213> Mouse <400> 222 ggtatctcac ttggtaggaa cctc 24 .
<210> 223 <211> 17 <212> DNA
<213> Mouse <400> 223 aagaacgctt ggctggc 17 <210> 224 <211> 20 <212> DNA
<213> Mouse <400> 224 gccgatcctg gtgatgtact 20 <210> 225 <211> 20 <212> DNA
<213> Mouse <400> 225 acaatggctc aaaaccgttc 20 <210> 226 <211> 20 <212> DNA
<213> Mouse <400> 226 gccttgggaa tttaccacct 20 <210> 227 <211> 20 <212> DNA
<213> Mouse <400> 227 agtacatcac caggatcggc 20 <210> 228 <211> 20 <212> DNA
<213> Mouse <400> 228 taaaaggcca tgcgataagc 20 <210> 229 <211> 20 <212> DNA
<213> Mouse <400> 229 agagctctgt ggggttctca 20 <210> 230 <211> 20 <212> DNA
<213> Mouse <400> 230 gaaggggaca gtgttggaga 20 <210> 231 <211> 20 <212> DNA
<213> Mouse <400> 231 tccatcaagg aaggatccac~ 20 <210> 232 <211> 19 <212> DNA
<213> Mouse <400> 232 ggtgggtaat gattggact 19 <210> 233 <211> 19 <212> DNA
<213> Mouse <400> 233 tgacgtggag ggaactgcc 19 <210> 234 <211> 20 <212> DNA
<213> Mouse <400> 234 tgagatctgg tgccctctct 20 <210> 235 <211> 20 <212> DNA
<213> Mouse <400> 235 gcctgatcta ggctggaaaa 20 <210> 236 <211> 20 <212> DNA
<213> Mouse <400> 236 aggcagaaag cagacaagga 20 <210> 237 <211> 20 <212> DNA
<213> Mouse <400> 237 cgacagcact tgtgaccact 20 <210> 238 <211> 20 <212> DNA
<213> Mouse <400> 238 ctgcagatgt agaccaggca 20 <210> 239 <211> 20 <212> DNA
<213> Mouse <400> 239 ctgtggtgga ttggacagtg 20 <210> 240 <211> 20 <212> DNA
<213> Mouse <400> 240 ttgcctaaca ctcccaaacc 20 <210> 241 <211> 20 <212> DNA
<213> Mouse <400> 241 tattaggagc accaccaggc 20 <210> 242 <211> 20 <212> DNA
<213> Mouse <400> 242 acctgtcttg tgggtggaag 20 <210> 243 <211> 20 <212> DNA
<213> Mouse <400> 243 ctgtggtgga ttggacagtg 20 <210> 244 <211> 20 <212> DNA
<213> Mouse <400> 244 gtggcttggt gctattgaca 20 <210> 245 <211> 20 <212> DNA
<213> Mouse <400> 245 ggggctatta aggccatttt 20 <210> 246 <211> 21 <212> DNA
<213> Mouse <400> 246 caattgagga atggctacca a ~ 21 <210> 247 <211> 20 <212> DNA
<213> Mouse <400> 247 tggcttcatg tccattgtgt 20 <210> 248 <211> 22 <212> DNA
<213> Mouse <400> 248 cagaaccaca aaggtaaatt gc 22 <210> 249 <211> 21 <212> DNA
<213> Mouse <400> 249 tcatgtttgc tgtccagttt g 21 <210> 250 <211> 29 <212> DNA
<213> Homo Sapiens <400> 250 gccaccatgc tgggcc.ctgc tgtcctggg 29 <210> 251 <211> 24 <212> DNA
<213> Homo Sapiens <400> 251 tcactcatgt ttcccctgat ttcc 24 <210> 252 <211> 20 <212> DNA
<213> Homo Sapiens <400> 252 ctgatttcct gtgttcccgt 20 <210>253 <211>20 <212>DNA

<213>Homo sapiens <400> 253 catgctggcc tacttcatca 20 <210> 254 <211> 29 <212> DNA
<213> Homo Sapiens <400> 254 gccttgcagg tcagctacgg tgctagcat 29 <210> 255 <211> 24 <212> DNA
<213> Homo Sapiens <400> 255 tcactcatgt ttcccctgat ttcc 24 <210> 256 <211> 20 <212> DNA
<213> Homo Sapiens <400> 256 aggaagcaga gaaaggccag 20 <210> 257 <211> 20 <212> DNA
<213> Homo Sapiens <400> 257 tcagaactgc ctctgagctg 20 <210> 258 <211> 20 <212> DNA
<213> Homo Sapiens <400> 258 tcttcacgta ctgggggaac 20 <210> 259 <211> 20 <212> DNA
<213> Homo sapiens <400> 259 actacagcat cagcagcagg 20 <210>260 <211>20 <212>DNA

<213>Homo sapiens <400> 260 aagctgaaga acttcccggt 20 <210> 261 <211> 20 <212> DNA
<213> Homo Sapiens <400> 261 tgggctacga cctctttgat 20 <210> 262 <211> 20 <212> DNA
<213> Homo Sapiens <400> 262 atcttcaggc gctctgtcct 20 <210>263 <211>20 <212>DNA

<213>Homo Sapiens <400> 263 gtacgacctg aagctgtggg 20 <210> 264 <211> 19 <212> DNA
<213> Homo Sapiens <400> 264 atcttcaggc gctctgtcc 19 <210> 265 <211> 20 <212> DNA
<213> Homo Sapiens <400> 265 gtacgacctg aagctgtggg 20 <210> 266 <211> 19 <212> DNA
<213> Homo Sapiens <400> 266 atcttcaggc gctctgtcc 19 <210> 267 <211> 21 <212> DNA
<213> Homo Sapiens <400> 267 gagtacgacc tgaagctgtg g 21 <210> 268 <211> 19 <212> DNA
<213> Homo Sapiens <400> 268 atcttcaggc gctctgtcc 19 <210> 269 <211> 19 <212> DNA
<213> Homo Sapiens <400> 269 tacgacctga agctgtggg 19 <210> 270 <211> 19 <212> DNA
<213> Homo Sapiens <400> 270 .
atcttcaggc gctctgtcc 19 <210> 271 <211> 19 <212> DNA
<213> Homo Sapiens <400> 271 tacgacctga agctgtggg 19 <210> 272 <211> 18 <212> DNA
<213> Homo Sapiens <400> 272 gctgtcccga tggtgaac 18 <210>273 <211>19 <212>DNA

<213>Homo Sapiens <400> 273 accttttgtg gccaggatg 19 <210>274 <211>18 <212>DNA

<213>Homo Sapiens <400> 274 gctgtcccga tggtgaac 18 <210>275 <211>19 <212DNA
>

<213>Homo Sapiens <400> 275 caccttttgt ggccaggat 19 <210> 276 <211> 18 <212> DNA
<213> Homo Sapiens <400> 276 gctgtcccga tggtgaac 18 <210> 277 <211> 18 <212> DNA
<213> Homo Sapiens <400> 277 ccttttgtgg ccaggatg 18 <210> 278 <211> 18 <212> DNA
<213> Homo Sapiens <400> 278 cctgaaccag tgggctgt 18 <210> 279 <211> 19 <212> DNA
<213> Homo Sapiens <400> 279 accttttgtg gccaggatg 19 <210> 280 <211> 18 <212> DNA
<213> Homo Sapiens <400> 280 cctgaaccag tgggctgt 18 <210> 281 <211> 19 <212> DNA
<213> Homo Sapiens <400> 281 caccttttgt ggccaggat 19 <210> 282 <211> 20 <212> DNA
<213> Homo Sapiens <400> 282 tcatgtttcc cctgatttcc 20 <210> 283 <211> 20 <212> DNA
<213> Homo Sapiens <400> 283 catgctggcc tacttcatca 20 <210> 284 <211> 20 <212> DNA
<213> Homo sapiens <400> 284 atgagcaggt aacacctggg 20 <210> 285 <211> 20 <212> DNA
<213> Homo sapiens <400> 285 tcatcacctg ggtctccttt 20 <210> 286 <211> 20 <212> DNA
<213> Homo Sapiens <400> 286 atgagcaggt aacacctggg 20 <210> 287 <211> 20 <212> DNA
<213> Homo Sapiens <400> 287 ttcatcacct gggtctcctt 20 <210> 288 <211> 20 <212> DNA
<213> Mouse <400> 288 tgggttgtgt tctctggttg 20 <210> 289 <211> 21 <212> DNA
<213> Mouse <400> 289 cctttttaca gtctgccagg t 21 <210> 290 <211> 20 <212> DNA
<213> Mouse <400> 290 tgggttgtgt tctctggttg 20 <210> 291 <211> 21 <212> DNA
<213> Mouse <400> 291 gatccccttt ttacagtctg c 21 <210> 292 <211> 20 <212> DNA
<213> Mouse <400> 292 acggggttgg tactgtgtgt 20 <210> 293 <211> 20 <212> DNA
<213> Mouse <400> 293 cacccattgt tagtgctgga 20 <210> 294 <211> 20 <212> DNA
<213> Mouse <400> 294 acggggttgg tactgtgtgt 20 <210> 295 <211> 20 <212> DNA
<213> Mouse <400> 295 cacacaccca cccattgtta 20 <210> 296 <211> 20 <212> DNA
<213> Mouse <400> 296 tgcattggcc agactagaaa 20 <210> 297 <211> 19 <212> DNA
<213> Mouse <400> 297 cggctgggct atgacctat 19 <210> 298 <211> 20 <212> DNA
<213> Mouse <400> 298 tgcattggcc agactagaaa 20 <210> 299 <211> 20 <212> DNA
<213> Mouse <400> 299 cggctgggct atgacctatt 20 <210> 300 <211> 20 <212> DNA
<213> Mouse <400> 300 gttctgcagc atgatgtcgt 20 <210> 301 <211> 20 <212> DNA
<213> Mouse <400> 301 _ ggcagttgtg actctgttgc 20 <210> 302 <211> 20 <212> DNA
<213> Mouse <400> 302 gttctgcagc atgatgtcgt 20 <210> 303 <211> 20 <212> DNA
<213> Mouse <400> 303 ctgcaggcag ttgtgactct 20 <210> 304 <211> 20 <212> DNA
<213> Mouse <400> 304 ccatcctttt tgcctgtctt 20 <210> 305 <211> 20 <212> DNA
<213> Mouse <400> 305 tctggaggaa catgtgatgg 20 <210> 306 <211> 20 <212> DNA
<213> Mouse <400> 306 caccatcctt tttgcctgtc 20 <210> 307 <211> 19 <212> DNA
<213> Mouse <400> 307 gaacatgtga tggggcaac 19 <210> 308 <211> 19 <212> DNA
<213> Mouse <400> 308 caaagcagca ggaggagtg 19 <210> 309 <211> 20 <212> DNA
<213> Mouse <400> 309 aaatgtactg gccaggcaac 20 <210> 310 <211> 20 <212> DNA
<213> Mouse <400> 310 agtgctagac ccagcaccag ~ 20 <210> 311 <211> 20 <212> DNA
<213> Mouse <400> 311 aaatgtactg gccaggcaac 20 <210> 312 <211> 20 <212> DNA
<213> Mouse <400> 312 gcactgacca gtctgtcacc 20 <210> 313 <211> 20 <212> DNA
<213> Mouse <400> 313 gtccccagag aaaagcacag 20 <210> 314 <211> 20 <212> DNA
<213> Mouse <400> 314 cagtctgtca ccacctctgg 20 <210> 315 <211> 20 <212> DNA
<213> Mouse <400> 315 cagtggtccc cagagaaaag 20 <210> 316 <211> 20 <212> DNA
<213> Mouse <400> 316 tactattcgg ggcttgttgg 20 <210> 317 <211> 20 <212> DNA
<213> Mouse <400> 317 gcagcactat gtgcctggta 20 <210> 318 <211> 20 <212> DNA
<213> Mouse <400> 318 tactattcgg ggcttgttgg 20 <210> 319 <211> 20 <212> DNA
<213> Mouse <400> 319 gcctggtatt tgatcgcttt 20 <210> 320 <211> 20 <212> DNA
<213> Mouse <400> 320 gctcagctag ggatggagaa 20 <210> 321 <211> 20 <212> DNA
<213> Mouse <400> 321 cagctcaggg acacaatgaa 20 <210> 322 <211> 20 <212> DNA
<213> Mouse <400> 322 tcctacaggc tagggctcag 20 <210> 323 <211> 20 <212> DNA
<213> Mouse <400> 323 cagctcaggg acacaatgaa 20 <210> 324 <211> 20 <212> DNA
<213> Mouse <400> 324 gggactgatg tgtggcttgt 20 <210> 325 <211> 20 <212> DNA
<213> Mouse <400> 325 aggcgtccca ggaatagaag 20 <210> 326 <211> 21 <212> DNA
<213> Mouse <400> 326 ggactgatgt gtggcttgtt t 21 <210> 327 <211> 20 <212> DNA
<213> Mouse <400> 327 aggcgtccca ggaatagaag 20 <210> 328 <211> 20 <212> DNA
<213> Mouse <400> 328 , tgtttctgtt ctggtggctg 20 <210> 329 <211> 20 <212> DNA
<213> Mouse <400> 329 atctgcaggc aggatcagac 20 <210> 330 <211> 20 <212> DNA
<213> Mouse <400> 330 ctcagtggtg ggtgacagtg 20 <210> 331 <211> 20 <212> DNA
<213> Mouse <400> 331 atctgcaggc aggatcagac 20 <210> 332 <211> 20 <212> DNA
<213> Mouse <400> 332 acacacagta ccaaccccgt 20 <210> 333 <211> 20 <212> DNA
<213> Mouse <400> 333 cctgtggtga tcaagaagca 20 <210> 334 <211> 20 <212> DNA
<213> Mouse <400> 334 tgcttcttga tcaccacagg 20 <210> 335 <211> 20 <212> DNA
<213> Mouse <400> 335 gcaacagagt cacaactgcc 20 <210> 336 <211> 20 <212> DNA
<213> Mouse <400> 336 acacacagta ccaaccccgt 20 <210> 337 <211> 20 <212> DNA
<213> Mouse <400> 337 gcaacagagt cacaactgcc 20 <210> 338 <211> 20 <212> DNA
<213> Mouse <400> 338 gggtttatgt ggcaagcact 20 <210> 339 <211> 20 <212> DNA
<213> Mouse <400> 339 actccatttg ccttttgtgg 20 <210> 340 <211> 20 <212> DNA
<213> Mouse <400> 340 cgctacttcg cttttatccg 20 <210> 341 <211> 20 <212> DNA
<213> Mouse <400> 341 atgatgacgt acgacgacga 20 <210> 342 <211> 21 <212> DNA
<213> Mouse <400> 342 gaaaacaatc ggggagaagt.c 21 <210> 343 <211> 20 <212> DNA
<213> Mouse <400> 343 tgaaattatc acacgccagg 20 <210> 344 <211> 20 <212> DNA
<213> Mouse <400> 344 agtgagaggc ccagtctcaa 20 <210> 345 <211> 20 <212> DNA
<213> Mouse <400> 345 gatctgatgc cctcttctgc 20 <210> 346 <211> 20 <212> DNA
<213> Mouse <400> 346 gctagccttg aagccaacac 20 <210> 347 <21~1> 20 <212> DNA
<213> Mouse <400> 347 tgaacagcat gcttacccag 20 <210> 348 <211> 20 <212> DNA
<213> Mouse <400> 348 tccctagagg cctgtctgtc 20 <210> 349 <211> 20 <212> DNA
<213> Mouse <400> 349 tcgtctcgga gcctcttcta 20 <210> 350 <211> 20 <212> DNA
<213> Mouse <400> 350 gatagtccct tagccagccc 20 <210> 351 <211> 20 <212> DNA
<213> Mouse <400> 351 gccatagctc ctcactgctc 20 <210> 352 <211> 20 <212> DNA
<213> Mouse <400> 352 cagagtgggc tctggtcttc 20 <210> 353 <211> 20 <212> DNA
<213> Mouse <400> 353 ttgtgttcag atgctcctgc 20 <210> 354 <211> 20 <212> DNA
<213> Mouse <400> 354 ttatttctgt gctagccgcc 20 <210> 355 <211> 20 <212> DNA
<213> Mouse <400> 355 atcaagtcaa cgtccccaag 20 <210> 356 <211> 20 <212> DNA
<213> Mouse <400> 356 acctggcctg tgctaatctc 20 <210> 357 <211> 20 <212> DNA
<213> Mouse <400> 357 gcaccaaccc taagaaagca 20 <210> 358 <211> 22 <212> DNA
<213> Mouse <400> 358 tcaggctaac ctcaaactca ca 22 <210> 359 <211> 27 <212> DNA
<213> Mouse <400> 359 aaagaaaaga aaagaaaaag tcagaca 27 <210> 360 <211> 20 <212> DNA
<213> Mouse <400> 360 cccagaactc catcctcaaa 20 <210> 361 <211> 20 <212> DNA
<213> Mouse <400> 361 cccaacctgt ggtcagctat 20 <210> 362 <211> 20 <212> DNA
<213> Mouse <400> 362 ggggcaggtg ggtaataagt 20 <210> 363 <211> 20 <212> DNA
<213> Mouse <400> 363 caaaagccca actccttgag 20 <210> 364 <211> 20 <212> DNA
<213> Mouse <400> 364 gctcagtggg taagagcac.c 20 <210> 365 <211> 20 <212> DNA
<213> Mouse <400> 365 ctaccctgcc gctaatctca 20 <210> 366 <211> 20 <212> DNA
<213> Mouse <400> 366 cagttagcac cccaccctaa 20 <210> 367 <211> 20 <212> DNA
<213> Mouse a.
<400> 367 tctgcacctc tgttcacctg 20 <210> 368 <211> 20 <212> DNA
<213> Mouse <400> 368 acctctaggg tttacgggga 20 <210> 369 <211> 20 <212> DNA
<213> Mouse <400> 369 cctcaggtag tgcaagctcc 20 <210> 370 <211> 20 <212> DNA
<213> Mouse <400> 370 tcagttacca agggtttcgg 20 <210> 371 <211> 20 <212> DNA
<213> Mouse <400> 371 ataggttgtc acaggccagg 20 <210> 372 <211> 20 <212> DNA
<213> Mouse <400> 372 tcagttacca agggtttcgg 20 <210> 373 <211> 20 <212> DNA
<213> Mouse <400> 373 ataggttgtc acaggccagg 20 <210> 374 <211> 20 <212> DNA
<213> Mouse <400> 374 gtggttgctg ggatttgaac 20 <210>375 <211>20 <212>DNA

<213>Mouse <400> 375 caagcaacca aacaaccaaa 20 <210> 376 <211> 20 <212> DNA
<213> Mouse <400> 376 tccggaggac cataaatctg 20 <210> 377 <211> 20 <212> DNA
<213> Mouse <400> 377 cacagtccca gtcattccct 20 <210> 378 <211> 20 <212> DNA
<213> Mouse <400> 378 gtcccaaaag ctagcacagg 20 <210> 379 <211> 20 <212> DNA
<213> Mouse <400> 379 tcatgagcca ccatgtgatt 20 <210> 380 <211> 20 <212> DNA
<213> Mouse <400> 380 gaccttcgga agagcagttg 20 <210> 381 <211> ~20 <212> DNA
<213> Mouse <400> 381 agtgtgtgtc gccatatcca 20 <210> 382 <211> 20 <212> DNA
<213> Mouse <400> 382 cctactctct ctccccgctt 20 <210> 383 <211> 20 <212> DNA
<213> Mouse <400> 383 ggaaaatgtt tggccttgaa 20 <210> 384 <211> 20 <212> DNA
<213> Mouse <400> 384 ctggagtgaa aggcaggaag 20 <210> 385 <211> 20 <212> DNA
<213> Mouse <400> 385 aggcggcacc atatgaataa 20 <210> 386 <211> 21 <212> DNA
<213> Mouse <400> 386 tgagagtggg aattctgttc a 21 <210> 387 <211> 20 <212> DNA
<213> Mouse <400> 387 ggatgtaatt ggtggcaagg 20 <210> 388 <211> 20 <212> DNA
<213> Mouse <400> 388 ctgttggagg aggtggccta 20 <210> 389 <211> 21 <212> DNA
<213> Mouse <400> 389 tgcttgtatg tttttcctcg t 21 <210> 390 <211> 20 <212> DNA
<213> Mouse <400> 390 tgagagtgcc ctcctctttg 20 <210> 391 <211> 18 <212> DNA
<213> Mouse <400> 391 gaacccctga ccccagac 18 <210> 392 <211> 22 <212> DNA
<213> Mouse <400> 392 tgaagtgcag atttttacat gg 22 <210> 393 <211> 20 <212> DNA
<213> Mouse <400> 393 gttttggggt ggaaaaggat 20 <210> 394 <211> 20 <212> DNA
<213> Mouse <400> 394 ccgtcgacat ttaggtgaca 20 <210> 395 <211> 20 <212> DNA
<213> Mouse <400> 395 gatactgggg tggtgggtaa 20 <210> 396 <211> 20 <212> DNA
<213> Mouse <400> 396 ccgtcgacat ttaggtgaca 20 <210> 397 <211> 20 <212> DNA
<213> Mouse <400> 397 cgtcccagct gtgtaactga 20 <210> 398 <211> 21 <212> DNA
<213> Mouse <400> 398 ggaagcaaat gctccactaa a 21 <210> 399 <211> 20 <212> DNA
<213> Mouse <400> 399 tatccctagc cccttgtgtg 20 <210> 400 <211> 20 <212> DNA
<213> Mouse <400> 400 ccgtcgacat ttaggtgaca 20 <210> 401 <211> 20 <212> DNA
<213> Mouse <400> 401 gggtcctgtt ggtagtgacc 20 <210> 402 <211> 20 <212> DNA
<213> Mouse <400> 402 tataagcagc ccctcattgg 20 <210> 403 <211> 20 <212> DNA
<213> Mouse <400> 403 caggccagac actgcttaca 20 <210> 404 <211> 20 <212> DNA
<213> Mouse <400> 404 ccttgggatc tggtgtgact 20 <210> 405 <211> 20 <212> DNA
<213> Mouse <400> 405 tgggtttaga gtacggctgg 20 <210> 406 <211> 20 <212> DNA
<213> Mouse <400> 406 acccatttcc taatcccctg 20 <210> 407 <211> 20 <212> DNA
<213> Mouse <400> 407 atctctccag cccctctcag 20 <210> 408 <211> 20 <212> DNA
<213> Mouse <400> 408 gggctgggaa ttgaacctat 20 <210> 409 <211> 20 <212> DNA
<213> Mouse <400> 409 tgaatccctt acagccttgc 20 <210> 410 <211> 20 <212> DNA
<213> Mouse <400> 410 gccccataaa atccactcct 20 <210> 411 <211> 20 <212> DNA
<213> Mouse <400> 411 gctccggaag gctagaagat 20 <210> 412 <211> 20 <212> DNA
<213> Mouse <400> 412 ggtttgggag tgttaggcaa 20 <210> 413 <211> 20 <212> DNA
<213> Mouse <400> 413 actcagttgg cctctcctca 20 <210> 414 <211> 19 <212> DNA
<213> Mouse <400> 414 acagaaatcc ctcatgcga 19 <210> 415 <211> 21 <212 > DNA
<213> Mouse <400> 415 tcagtgtgga ccagaaagtc c 21 <210>416 <211>22 <212>DNA

<213>Mouse <400> 416 tctgcaagtc agctcttgat as 22 <210> 417 <211> 23 <212> DNA
<213> Mouse <400> 417 actcataagg gtcaagctgt ctg ~ 23 <210> 418 <211> 20 <212> DNA
<213> Mouse <400> 418 tctccccttt taccactccc 20 <210> 419 <211> 20 <212> DNA
<213> Mouse <400> 419 gcaaggagtc aaaaacagca 20 <210> 420 <211> 20 <212> DNA
<213> Mouse <400> 420 gctagttggg gaacaaacca 20 <210> 421 <211> 20 <212> DNA
<213> Mouse <400> 421 actgcaaatg tccaactcca 20 <210> 422 <211> 20 <212> DNA
<213> Mouse <400> 422 cagttacaca gctgggacga 20 <210> 423 <211> 20 <212> DNA
<213> Mouse <400> 423 gcaagagcct agcaatccac 20 <210> 424 <211> 20 <212> DNA
<213> Mouse <400> 424 cagtttagca ccccacccta 20 <210> 425 <211> 20 <212> DNA
<213> Mouse <400> 425 tctgcacctc tgttcacctg 20 <210> 426 <211> 20 <212> DNA
<213> Mouse <400> 426 gggttccact tgatgctgat 20 <210> 427 <211> 20 <212> DNA
<213> Mouse <400> 427 tggtctgttt cctggagctt 20 <210> 428 <211> 21 <212> DNA
<213> Mouse <400> 428 tgtagggaat gtttctgcac c 21 <210> 429 <211> 20 <212> DNA
<213> Mouse <400> 429 acatggaaca ggattctggc 20 <210> 430 <211> 20 <212> DNA
<213> Mouse <400> 430 gcaggcaaac agacagacaa 20 <210> 431 <211> 20 <212> DNA
<213> Mouse <400> 431 atgggggatc ccttactgac 20 <210> 432 <211> 20 <212> DNA
<213> Mouse <400> 432 cggtcaggag tagtgtgggt 20 <210> 433 <211> 20 <212> DNA
<213> Mouse <400> 433 cagcagctga tattgaggca 20 <210> 434 <211> 22 <212> DNA
<213> Mouse <400> 434 aatgatgaag tgtcagcctc ag 22 <210> 435 <211> 20 <212> DNA
<213> Mouse <400> 435 caacagaact caaagcctgg 20 <210> 436 <211> 20 <212> DNA
<213> Mouse <400> 436 agcaggcaca ggtctcttgt 20 <210> 437 <211> 20 <212> DNA
<213> Mouse <400> 437 aagaacagga cagtggtggg 20 <210> 438 <211> 20 <212> DNA
<213> Mouse <400> 438 cagcgattgg ctcttctctt 20 <210> 439 <211> 20 <212> DNA
<213> Mouse <400> 439 ggggcttcct ttctgaggta 20 <210> 440 <211> 20 <212> DNA
<213> Mouse <400> 440 agctcaggtc cagcttggta 20 <210> 441 <211> 20 <212> DNA
<213> Mouse <400> 441 attttcccct cctgcttctc 20 <210> 442 <211> 20 <212> DNA
<213> Mouse <400> 442 ccaagcctct gctggttatc 20 <210> 443 <211> 20 <212> DNA
<213> Mouse <400> 443 tgagggtgga gaatggaaag 20 <210> 444 <211> 20 <212> DNA
<213> Mouse <400> 444 gccccataaa atccactcct 20 <210> 445 <211> 20 <212> DNA
<213> Mouse <400> 445 ttgcctaaca ctcccaaacc 20 <210> 446 <211> 20 <212> DNA
<213> Mouse <400> 446 cagttacaca gctgggacga 20 <210> 447 <211> 20 <212> DNA
<213> Mouse <400> 447 gcaagagcct agcaatccac 20 <210> 448 <211> 20 <212> DNA
<213> Mouse <400> 448 cagcaccttc ctctggtctc 20 <210> 449 <211> 20 <212> DNA
<213> Mouse <400> 449 tgtctccaga ggttctgcct 20 <210> 450 <211> 24 <212> DNA
<213> Mouse <400> 450 tggtggtgta atactattcc tttg 24 <210> 451 <211> 26 <212> DNA
<213> Mouse <400> 451 tctttaattt ttggcttttt gataca 26 <210> 452 <211> 20 <212> DNA
<213> Mouse <400> 452 cagctgtgtg catgttgacc 20 <210> 453 <211> 20 <212> DNA
<213> Mouse <400> 453 catcatgaag actcagggca 20 <210> 454 <211> 20 <212> DNA
<213> Mouse <400> 454 gtccacacct ggcttttgtt 20 <210> 455 <211> 20 <212> DNA
<213> Mouse <400> 455 cagcactcag tgaggttcca 20 <210> 456 <211> 20 <212> DNA
<213> Mouse <400> 456 atgtaatgga agggctgctg 20 <210> 457 <211> 20 <212> DNA
<213> Mouse <400> 457 cagcactcag tgaggttcca 20 <210> 458 <211> 21 <212> DNA
<213> Mouse <400> 458 aaacaggcat gaaactcagg a 21 <210> 459 <211>~ 20 <212> DNA
<213> Mouse <400> 459 gggtatcatt gtcacctcca 20 <210> 460 <211> 20 <212> DNA
<213> Mouse <400> 460 cacaggccaa gttgttgttg 20 <210> 461 <211> 20 <212> DNA
<213> Mouse <400> 461 caggggacct tctgaatgat 20 <210> 462 <211> 20 <212> DNA
<213> Mouse <400> 462 agctcaggtc cagcttggta 20 <210> 463 <211> 20 <212> DNA
<213> Mouse <400> 463 accacaaaat tttcccctcc 20 <210> 464 <211> 20 <212> DNA
<213> Mouse <400> 464 cgggacctaa aactggacaa 20 <210> 465 <211> 20 <212> DNA
<213> Mouse <400> 465 tggggacagt taccaggaag 20 <210> 466 <211> 20 <212> DNA
<213> Mouse <400> 466 ccggaggacc ataaatctga 20 <210> 467 <211> 20 <212> DNA
<213> Mouse <400> 467 cctcaaaaac aagcctgagc 20 <210> 468 <211> 22 <212> DNA
<213> Mouse <400> 468 ccttcagaaa tgtgtttgga ca 22 <210> 469 <211> 20 <212> DNA
<213> Mouse <400> 469 tcctgagttc aaatcccagc 20 <210> 470 <211> 20 <212> DNA
<213> Mouse <400> 470 ctttccattc tccaccctca 20 <210> 471 <211> 20 <212> DNA
<213> Mouse <400> 471 aggtcctagg gagaggtcca 20 <210> 472 <211> 20 <212> DNA
<213> Mouse <400> 472 aggcctaccc aaggacatct 20 <210> 473 <211> 20 <212> DNA
<213> Mouse <400> 473 gcagtgagct gcagagtttg ~ 20 <210> 474 <211> 20 <212> DNA
<213> Mouse <400> 474 agacacccta ggtcctgctg 20 <210> 475 <211> 22 <212> DNA
<213> Mouse <400> 475 tgatctttcc aaacgcataa ga 22 <210> 476 <211> 20 <212> DNA
<213> Mouse <400> 476 gcaagcaacc tgaacatgaa 20 <210> 477 <211> 20 <212> DNA
<213> Mouse <400> 477 gcttacgatg gtcgtgaggt 20 <210> 478 <211> 20 <212> DNA
<213> Mouse <400> 478 acatgcctgc ctatctttgc 20 <210> 479 <211> 20 <212> DNA
<213> Mouse <400> 479 ggaacctgtt ttccatggtg . 20 <210> 480 <211> 20 <212> DNA
<213> Mouse <400> 480 accttgttcc tggtgtgagc 20 <210> 481 <211> 20 <212> DNA
<213> Mouse <400> 481 tagctgggac gtggtatggt 20 <210> 482 <211> 20 <212> DNA
<213> Mouse <400> 482 ccatgggaga ccagaaggta 20 <210> 483 <211> 20 <212> DNA
<213> Mouse <400> 483 tgagtgtcct ctgcctgatg 20 <210> 484 <211> 20 <212> DNA
<213> Mouse <400> 484 gcgctgacat cctcctatgt 20 <210> 485 <211> 20 <212> DNA
<213> Mouse <400> 485 cccactatgg tcccagagaa 20 <210> 486 <211> 20 <212> DNA
<213> Mouse <400> 486 ttgcacgtct ttgtttcgag 20 <210> 487 <211> 24 <212> DNA
<213> Mouse <400> 487 aaaggggaat agacctgagt agaa 24 <210> 488 <211> 20 <212> DNA
<213> Mouse <400> 488 ccaagagtca gccttggagt 20 <210> 489 <211> 20 <212> DNA
<213> Mouse <400> 489 ggacaggtag ctcacccaac 20 <210> 490 <211> 19 <212> DNA
<213> Mouse <400> 490 tgccagcttt ggctatcat 19 <210> 491 <211>~20 <212> DNA
<213> Mouse <400> 491 ttcattgtgt ccctgagctg 20 <210> 492 <211> 24 <212> DNA
<213> Mouse <400> 492 agctttggct atcatgggtc tcag 24 <210> 493 <211> 22 <212> DNA
<213> Mouse <400> 493 accaccgcca ctgttctcat ct 22 <210> 494 <211> 20 <212> DNA
<213> Mouse <400> 494 tgtgggggaa gaacatagaa 20 <210> 495 <211> 22 <212> DNA
<213> Mouse <400> 495 tgatgtgtgg cttgtttctc tt 22 <210> 496 <211> 20 <212> DNA
<213> Mouse <400> 496 ataggtgggg agggagctaa 20 <210> 497 <211> 22 <212> DNA
<213> Mouse <400> 497 tgatgtgtgg cttgtttctc tt 22 <210> 498 <211> 20 <212> DNA
<213> Homo Sapiens <400> 498 tgtgcctgtc acagcaactt 20 <210> 499 <211> 20 <212> DNA
<213> Homo Sapiens <400> 499 catgctagca ccgtagctga 20 <210> 500 <211> 20 <212> DNA
<213> Homo Sapiens <400> 500 ggagaccttc ccctccttct 20 <210> 501 <211> 20 <212> DNA
<213> Homo Sapiens <400> 501 gctgtagttg aagagggcgt 20 <210> 502 <211> 18 <212> DNA
<213> Homo Sapiens <400> 502 gtgcttggct tcctccag 18 <210> 503 <211> 20 <212> DNA
<213> Homo Sapiens <400> 503 caggtcgtac tccatgtcca 20 <210> 504 <211> 20 <212> DNA
<213> Homo Sapiens <400> 504 tggagtacga cctgaagctg 20 <210>505 <211>20 <212>DNA

<213>Homo Sapiens <400> 505 actcatcctg gccacaaaag . 20 <210> 506 <211> 19 <212> DNA
<213> Homo Sapiens <400> 506 gaacaggagg acgctgagg 19 <210> 507 <211> 20 <212> DNA
<213> Homo Sapiens <400> 507 cttttgtggc caggatgagt 20 <210> 508 <211> 20 <212> DNA
<213> Homo Sapiens <400> 508 tcacctcacc tggttgtcag 20 <210> 509 <211> 20 <212> DNA
<213> Homo sapiens <400> 509 gtacgacctg aagctgtggg 20 <210> 510 <211> 27 <212> DNA
<213> Homo Sapiens <400> 510 ggctgagatc acagggttgg gtcactc 27 <210> 511 <211> 27 <212> DNA
<213> Homo Sapiens <400> 511 ccgtgcctgt tggaagttgc ctctgcc 27 <210> 512 <211> 20 <212> DNA
<213> Mouse <400> 512 aattcccagc aaccactcac 20 <210> 513 <211> 20 <212> DNA
<213> Mouse <400> 513 cagacactcc agaagagggc 20 <210> 514 <211> 20 <212> DNA
<213> Mouse <400> 514 tgactgctct tccgaaggtt 20 <210> 515 <211> 20 <212> DNA
<213> Mouse <400> 515 tttgtggaat agccaaagcc 20 <210> 516 <211> 20 <212> DNA
<213> Mouse <400> 516 tctctcctct cttctccccc 20 <210> 517 <211> 20 <212> DNA
<213> Mouse <400> 517 agcagggtgc atcaccttat 20 <210> 518 <211> 20 <212> DNA
<213> Mouse <400> 518 taggagtgcc ccataggttg 20 <210> 519 <211> 20 <212> DNA
<213> Mouse <400> 519 tcattgtacc cagccagtca 20 <210> 520 <211> 20 <212> DNA
<213> Mouse <400> 520 aggactgagc ctggatgaga 20 <210> 521 <211> 20 <212> DNA
<213> Mouse <400> 521 ctgggcgttt tgttttgttt 20 <210> 522 <211> 20 <212> DNA
<213> Mouse <400> 522 cttcctcctg cagctaccac 20 <210> 523 <211> 20 <212> DNA
<213> Mouse <400> 523 accctgctac aacgcagact 20 <210> 524 <211> 20 <212> DNA
<213> Mouse <400> 524 tccaaccttg acacccattt 20 <210> 525 <211> 20 <212> DNA
<213> Mouse <400> 525 agccagggct acacagagaa 20 <210> 526 <211> 20 <212> DNA
<213> Mouse <400> 526 ctgcttttcc tcagcaactg 20 <210> 527 <211> 20 <212> DNA
<213> Mouse <400> 527 attcgccgtt agaagctagg 20 <210> 528 <211> 20 <212> DNA
<213> Mouse P

<400> 528 aactgtacgt ggctgctggt 20 <210> 529 <211> 20 <212> DNA
<213> Mouse <400> 529 attcgccgtt agaagctagg 20 <210> 530 <211> 20 <212> DNA
<213> Mouse <400> 530 gccaggtgac ccttatgaaa 20 <210> 531 <211> 20 <212> DNA
<213> Mouse <400> 531 gagagatggc agacagaggc 20 <210> 532 <211> 20 <212> DNA
<213> Mouse <400> 532 agctctctgt ccctggtgaa 20 <210> 533 <211> 20 <212> DNA
<213> Mouse <400> 533 tgccaaccac tagcctctct 20 <210> 534 <211> 20 <212> DNA
<213> Mouse <400> 534 ctgaaccctc cactctcctg 20 <210> 535 <211> 20 <212> DNA
<213> Mouse <400> 535 agccagggct acacagagaa 20 <210> 536 <211> 20 <212> DNA
<213> Mouse <400> 536 agccagggct acacagagaa 20 <210> 537 <211> 20 <212> DNA
<213> Mouse <400> 537 accctgctac aacgcagact 20 <210> 538 <211> 20 <212> DNA
<213> Mouse <400> 538 gcaagtttca ggagctaggg 20 <210> 539 <211> 20 <212> DNA
<213> Mouse <400> 539 ccccagaacc agagaccata , 20 <210> 540 <211> 20 <212> DNA
<213> Mouse <400> 540 ccccagaacc agagaccata 20 <210> 541 <211> 20 <212> DNA
<213> Mouse <400> 541 ctaggggact ctgccaagtg 20 <210> 542 <211> 20 <212> DNA
<213> Mouse <400> 542 caagacaccc agtcccaact 20 <210> 543 <211> 20 <212> DNA
<213> Mouse <400> 543 tacttcccct ttcccgaact 20 <210> 544 <211> 20 <212> DNA
<213> Mouse <400> 544 tccttggtgc ttaccctcac 20 <210> 545 <211> 20 <212> DNA
<213> Mouse <400> 545 tgttcctgag ttcacaacgc 20 <210> 546 <211> 20 <212> DNA
<213> Mouse <400> 546 attcccagca actacatggc 20 <210> 547 <211> 20 <212> DNA
<213> Mouse <400> 547 acatgtccac tgtggcaaaa 20 <210> 548 <211> 20 <212> DNA
<213> Mouse <400> 548 tgtcatgagt ttgaggccag 20 <210> 549 <211> 20 <212> DNA
<213> Mouse <400> 549 atcagacagc ccacaacctc 20 <210> 550 <211> 20 <212> DNA
<213> Mouse <400> 550 tatgtgccac cacacctgtc 20 <210> 551 <211> 20 <212> DNA
<213> Mouse <400> 551 gctcaaggaa ggacacacct 20 <210> 552 <211> 22 <212> DNA
<213> Mouse <400> 552 tgctcttaac attttgagcc at 22 <210> 553 <211> 20 <212> DNA
<213> Mouse <400> 553 gctcagcccc tgaatcaata 20 <210> 554 <211> 20 <212> DNA
<213> Mouse <400> 554 gggatctgcc tgtcttacca 20 <210> 555 <211> 20 <212> DNA
<213> Mouse <400> 555 ggaaggtagg gcctggtaat 20 <210> 556 <211> 20 <212> DNA
<213> Mouse <400> 556 gctccaagat ctgtgcgatt 20 <210> 557 <211> 20 <212> DNA
<213> Mouse <400> 557 ttagcgttag ggtgagggtg 20 <210> 558 <211> 20 <212> DNA
<213> Mouse <400> 558 ggagactacg gacttgtggc 20 <210> 559 <211> 20 <212> DNA
<213> Mouse <400> 559 cagttcttcc cgaaaaccac 20 <210> 560 <211> 20 <212> DNA
<213> Mouse <400> 560 tttctgggaa ctgagatggc 20 <210> 561 <211> 20 <212> DNA
<213> Mouse <400> 561 gttggggctg ctcatagaaa 20 <210> 562 <211> 20 <212> DNA
<213> Mouse <400> 562 gctgtggctc tcttggagtt 20 <210> 563 <211> 20 <212> DNA
<213> Mouse <400> 563 ctctgatttc ccacatgcct 20 <210> 564 <211> 20 <212> DNA
<213> Mouse <400> 564 aagagggagc actgaggaca 20 <210> 565 <211> 20 <212> DNA
<213> Mouse <400> 565 cagcagcaaa tgacctttca 20 <210> 566 <211> 20 <212> DNA
<213> Mouse <400> 566 gaggcaggca gatttctgag 20 <210> 567 <211> 20 <212> DNA
<213> Mouse <400> 567 gtttcacatg ttgtggtggc 20 <210> 568 <211> 20 <212> DNA
<213> Mouse <400> 568 gggacctttg ggatagcatt 20 <210> 569 <211> 20 <212> DNA
<213> Mouse <400> 569 i tcagacatct ctggcctcct 20 <210>570 "

<211>20 <212>DNA

<213>Mouse <400> 570 ttcactaagt tgcccaggct 20 <210> 571 <211> 22 <212> DNA
<213> Mouse <400> 571 tgcctttttc tcacattgtc tc 22 <210> 572 <211> 20 <212> DNA
<213> Mouse <400> 572 ttagaagcag aggcagaggc 20 <210> 573 <211> 20 <212> DNA
<213> Mouse <400> 573 gacctttgga agagcagtcg 20 <210> 574 <211> 20 <212> DNA
<213> Mouse <400> 574 tggcagctca caatgtcttt 20 <210> 575 <211> 20 <212> DNA
<213> Mouse <400> 575 ggtgtggtgt aggggaagaa 20 <210> 576 <211> 22 <212> DNA
<213> Mouse <400> 576 tttcaactgc aaacacaaac ag 22 <210> 577 <211> 19 <212> DNA
<213> Mouse <400> 577 agggccaagg aaggagaat 19 <210> 578 <211> 24 <212> DNA
<213> Mouse <400> 578 gcaaatatat agggtaccga gctg 24 <210> 579 <211> 20 <212> DNA
<213> Mouse <400> 579 cagattctcc agctgtcagg 20 <210> 580 <211> 19 <212> DNA
<213> Mouse <400> 580 ctgtgtttcc gcaccaagt 19 <210> 581 <211> 20 <212> DNA
<213> Mouse <400> 581 ctgcccgtcc ttatcttctg 20 <220> 582 <211> 20 <212> DNA
<213> Mouse <400> 582 acgcacgctc actcatacac 20 <210> 583 <211> 20 <212> DNA
<213> Mouse <400> 583 cagcagaggt gatgggttct 20 <210> 584 <211> 22 <212> DNA
<213> Mouse <400> 584 ttgtcacaca gtggttaaat gc 22 <210> 585 <211> 20 <212> DNA
<213> Mouse <400> 585 tagaaccgtg gctgaggact 20 <210> 586 <211> 24 <212> DNA
<213> Mouse <400> 586 ccgtaagata tgaaagaact tgga 24 <210> 587 <211> 20 <212> DNA
<213> Mouse <400> 587 taatcctggc ttagcgcttg 20 <210> 588 <211> 20 <212> DNA
<213> Mouse <400> 588 tagaaagcac aggggacagg 20 <210> 589 <211> 20 <212> DNA
<213> Mouse <400> 589 ccttcctcgt ctgagctgtt 20 <210> 590 <211> 20 <212> DNA
<213> Mouse <400> 590 ttgggacgtg acctgagaat 20 <210> 591 <211> 20 <212> DNA
<213> Mouse <400> 591 tatgtgtctg gccgttgttc 20 <210> 592 <211> 19 <212> DNA
<213> Mouse <400> 592 gatgtgggtg caggtgaag 19 <210> 593 <211> 20 <212> DNA
<213> Mouse <400> 593 ccccttctgg agtgtctgaa 20 <210> 594 <211> 21 <212> DNA
<213> Mouse <400> 594 tctaggcagg gctacctttt t 21 <210> 595 <211> 19 <212> DNA
<213> Mouse <400> 595 gctgagcagc ctctagcaa 19 <210> 596 <211> 20 <212> DNA
<213> Mouse <400> 596 accatggctt ttcccagtaa 20 <210> 597 <211> 20 <212> DNA
<213> Mouse <400> 597 ctgtgccttt ggtgatcaga 20 <210> 598 <211> 20 <212> DNA
<213> Mouse <400> 598 tgtggcactc tacggcataa 20 <210> 599 <211> 23 <212> DNA
<213> Mouse <400> 599 tgcatcacta ttaagcctca acc 23 <210> 600 <211> 23 <212> DNA
<213> Mouse <400> 600 aagaatttgc aaagactgtg aga 23 <210> 601 <211> 20 <212> DNA
<213> Mouse <400> 601 ctggaccttt ggaagagcag 20 <210> 602 <211> 20 <212> DNA
<213> Mouse <400> 602 ggtggctcaa accatccata 20 <210> 603 <211> 20 <212> DNA
<213> Mouse <400> 603 gagggcaatg agcaaaatgt 20 <210> 604 <211> 20 <212> DNA
<213> Mouse <400> 604 ggtcctgtct ctggttcagg 20 <210> 605 <211> 20 <212> DNA
<213> Mouse <400> 605 taacacccac atcaggcaac 20 <210> 606 <211> 22 <212> DNA
<213> Mouse <400> 606 tttcatttcc tggtgttcct tt 22 <210> 607 <211> 20 <212> DNA
<213> Mouse <400> 607 aaacacaggc ggaacgatag 20 <210> 608 <211> 20 <212> DNA
<213> Mouse <400> 608 ctatcgttcc gcctgtgttt 20 <210> 609 <211> 21 <212> DNA
<213> Mouse <400> 609 aaggaagagg atggagaaag a 21 <210> 610 <211> 20 <212> DNA
<213> Mouse <400> 610 cgggtcttaa tggagcagag 20 <210> 611 <211> 20 <212> DNA
<213> Mouse <400> 611 tcctccccag ttacctagca 20 <210> 612 <211> 19 <2l2> DNA
<213> Mouse <400> 612 cagcaggcaa gatgacctc 19 <210> 613 <211> 20 <212> DNA
<213> Mouse <400> 613 gtccctcacc agccatgtta 20 <210> 614 <211> 20 <212> DNA
<213> Mouse <400> 614 agcctgggct aagttgtgtg 20 <210> 615 <211> 20 <212> DNA
<213> Mouse <400> 615 tatgggccaa tgttgttcct 20 <210> 616 <211> 20 <212> DNA
<213> Mouse <400> 616 atggtggctc acaaccatct 20 <210> 617 <211> 20 <212> DNA
<213> Mouse <400> 617 ttgtcctctg attgcagcat 20 <210> 618 <211> 20 <212> DNA
<213> Mouse <400> 618 cttgggtcat caggctttgt 20 <210> 619 <211> 20 <212> DNA
<213> Mouse <400> 619 aagctgccct gctctctctap 20 <210> 620 <211> 20 <212> DNA
<213> Mouse <400> 620 atgctcagcc tgctttgttt 20 <210> 621 <211> 20 <212> DNA
<213> Mouse <400> 621 gctgatagcc ctgggttcta 20 <210> 622 <211> 21 <212> DNA
<213> Mouse <400> 622 tgtacgcaca aattgacttg c 21 <210> 623 <211> 21 <212> DNA
<213> Mouse <400> 623 gaatccacat tgcaaagcct a 21 <210> 624 <211> 20 <212> DNA
<213> Mouse <400> 624 cacaggcaaa tgaagggaag 20 <210> 625 <211> 20 <212> DNA
<213> Mouse <400> 625 ccagacttct ccagctctcc 20 <210> 626 <211> 21 <212> DNA
<213> Mouse <400> 626 tcctcgagag gctctaggtt t 21 <210> 627 <211> 20 <212> DNA
<213> Mouse <400> 627 tgcctagtca accacaggag 20 <210> 628 <211> 21 <212> DNA
<213> Mouse <400> 628 v cctgtggttg actaggcaga a 21 <210> 629 <211> 20 <212> DNA
<213> Mouse <400> 629 gcctgatagc ctggaataca 20 <210> 630 <211> 20 <212> DNA
<213> Mouse <400> 630 aaagggatgt gtggcgtaag 20 <210> 631 <211> 20 l <212> DNA
<213> Mouse .
<400> 631 caaaacccaa ccttctcagc 20 <210> 632 <211> 20 <212> DNA
<213> Mouse <400> 632 tgcactgacc gtgatagagg 20 <210> 633 <211> 20 <212> DNA
<213 >_ Mouse <400> 633 eggtgtagct ctggctgtct 20 <210> 634 <211> 20 <212> DNA
<213> Mouse <400> 634 catctcacca actcgcactt 20 <210> 635 <211> 21 <212 > DNA
<213> Mouse <400> 635 tttctgggaa caaagaggct a 21 <210> 636 <211> 20 <212 > DNA
<213> Mouse <400> 636 gaacccaagt gttggggtaa 20 <210> 637 <211> 20 <212 > DNA
<213> Mouse <400> 637 tggaagccca tctgtctctt 20 <210> 638 <211> 20 <212 > DNA
<213> Mouse <400> 638 aaatgcaagt gggtgcttct 20 <210> 639 <211> 19 <212> DNA
<213> Mouse <400> 639 ccagaagagg gcgtcagat 19 <210> 640 <211> 20 <212> DNA
<213> Mouse <400> 640 ggtgtgcacc accatattca 20 <210> 641 <211> 21 <212> DNA
<213> Mouse <400> 641 gggaattatc agccaaaaag c 21 <210> 642 <211> 20 <212> DNA
<213> Mouse <400> 642 gcccaactga aagctcaact 20 <210> 643 <211> 21 <212> DNA
<213> Mouse <400> 643 ggaaggggga taacaattga a 21 <210> 644 <211> 23 <212> DNA
<213> Mouse <400> 644 tgctaatttc aagcacagtg aga 23 <210> 645 <211> 20 <212> DNA
<213> Mouse <400> 645 agcttgacac cttgacagca 20 <210> 646 <211> 20 <212> DNA
<213> Mouse <400> 646 aacctgcaga gaggagacca 20 <210> 647 <211> 20 <212> DNA
<213> Mouse <400> 647 ctccaagggg aggactcatt 20 <210> 648 <211> 24 <212> DNA
<213> Mouse <400> 648 ttcaattgag tttctctcct ctga 24 <210> 649 <211> 20 <212> DNA
<213> Mouse <400> 649 tgcaggacca agaagtaggc 20 <210> 650 <211> 20 <212> DNA
<213> Mouse <400> 650 cgagatctga tgccctcttc 20 <210> 651 <211> 20 <212> DNA
<213> Mouse <400> 651 tgctgagagc agaaaaggaa 20 <210> 652 <211> 166 <212> DNA
<213> Mouse <400> 652 gcagtgagct gcagagtttg cagaatgagg gcactctaaa ctcatcaagt gaggaggccc 60 ttccctcaca ctccagatgg ctgataggtg gcattacatg gtccancgcg cgcacgcgct 120 cagatgcaat ctccacattc ataaccagat gtccttgggt aggcct 166

Claims (38)

-98- What is claimed is:

1. An isolated polynucleotide comprising a sequence variation of SEQ ID.
NO 1, wherein said variation is associated with sensing carbohydrates, other sweeteners, or ethanol.

2. An isolated polynucleotide comprising a sequence variation of SEQ ID.
NO 2, wherein said variation is associated with sensing carbohydrates, other sweeteners, or ethanol.

3. An isolated polynucleotide comprising a sequence variation of SEQ ID.
NO 4, wherein said variation is associated with altered sensation of carbohydrates, other sweeteners, or ethanol.

4. The polynucleotide of Claim 1 wherein said variation is a missense mutation.

5. The polynucleotide of Claim 4 wherein said variation is a nonsense mutation.

6. An isolated polypeptide comprising a variant form of SEQ ID. NO: 3, wherein said variant form is associated with altered preference for carbohydrates, other sweeteners, or ethanol.

7. An isolated polypeptide comprising a variant form of SEQ ID. NO 5, wherein said variant form is associated with altered preference for carbohydrates, other sweeteners, or ethanol.

8. An isolated polynucleotide having at least 8 contiguous nucleotides of the polynucleotides of any one of the Claims 1-3 wherein said 8 contiguous nucleotides span said variation position.

9. An isolated polypeptide having at least four contiguous amino acids of the polypeptides of Claims 6 or 7 wherein said four contiguous amino acids span said variation position.

10. An isolated polynucleotide wherein said polynucleotide is substantially identical to the polynucleotide of Claim 8.

11. An isolated polypeptide wherein said polypeptide is substantially identical to the polypeptide of Claim 9.

12. An isolated polynucleotide having a sequence which is complementary to the polynucleotide of Claim 8 or 10.

13. A polynucleotide specific for the SAC1 locus wherein said polynucleotide hybridizes, under stringent conditions, to at least 8 contiguous nucleotides of the polynucleotide of Claim 1, 2, 3, or 4.

14. The polynucleotide according to Claim 13 wherein said polynucleotide is selected from the group consisting of SEQ ID. NOS 6-651 and homologous equivalents thereof.

15. A polynucleotide specific for the SAC1 locus wherein said polynucleotide that hybridizes, under stringent conditions, to at least 8 contiguous nucleotides of the polynucleotide of Claim 3.

16. The polynucleotide of Claim 15 wherein said polynucleotide is selected from the group consisting of SEQ ID. NOS 6-651 and homologous equivalents thereof.

17. A kit for the detection of the polynucleotide of any one of Claims 1-5, 8, and 10 comprising a polynucleotide that hybridizes, under stringent conditions, to at least 12 contiguous nucleotides of the polynucleotide of any one of the Claims 1-5, 8, and 10, and instructions relating to detection.

18. An isolated antibody which is immunoreactive to the polypeptide of Claim 9 or 11.

19. A method for analyzing a biomolecule in a biological sample, wherein said method comprising:
a) altering SAC1 activity in a biological sample; and b) measuring the activity.

20. A method for analyzing a polynucleotide in a biological sample comprising the steps of:
a) contacting a polynucleotide in a biological sample with a probe wherein said probe hybridizes to the polynucleotides of Claim 8 or 10 to form a hybridization complex; and b) detecting the hybridization complex.

21. A method for analyzing the expression of SAC1 comprising the steps of a) contacting a biological sample with a probe wherein said probe comprises the polynucleotide of Claim 8 or 10; and b) detecting the expression of SAC1 mRNA transcript in said sample.

22. The method of Claim 19 wherein said step of measuring is an enzymatic assay.

23. The method of Claim 20 or 21 wherein said probe is immobilized on a solid support.

24. The method according to any one of the Claims 19-23 wherein said sample is derived from blood.

25. The method according to any one of the Claims 19-23 wherein said sample is derived from tongue.

26. The method according to any one of the Claims 19-23 wherein said sample is derived from pancreas.

27. The method according to any one of the Claims 19-23 wherein said sample is derived from a human.

28. A method for identifying susceptibility to obesity or diabetes which comprises comparing the nucleotide sequence of the suspected SAC1 allele with a wild type nucleotide sequence, wherein said difference between the suspected allele and the wild-type sequence identifies a sequence variation of the SACl nucleotide sequence.

29. An expression vector comprising the polynucleotide of Claim 3, 8, or 10.

30. A host cell comprising the expression vector of Claim 29.

31. A method of producing a polypeptide comprising culturing the cells of Claim 30 and recovering the polypeptide from the host cell.

32. An isolated polypeptide produced according to Claim 31.

33. A method for conducting a screening assay to identify a molecule which enhances or decreases the SACl activity comprising the steps of a) contacting a biological sample with a molecule wherein said biological sample contains SACl activity; and b) analyzing the SAC1 activity in said sample.

34. A pharmaceutical composition comprising a) the polynucleotide of Claim 8 or 10, the polypeptide of Claim 9 or 11, the antibody of Claim 18 or the molecule of Claim 18; and b) a suitable pharmaceutical carrier.

35. A method for treating or preventing obesity, diabetes, or alcoholism associated with expression of SAC1, wherein said method comprises administering to a subject an effective amount of the pharmaceutical composition of Claim 34.

36. A transgenic animal that carries an altered SAC1 allele.

37. The transgenic animal of Claim 36 is a knock out mouse.

38. The polypeptide of Claim 6 or 7, wherein said polypeptide is 7-transmembrane G protein coupled receptor (7TM GPCR).