US2003187246A1PendingUtilityA1

Nucleic acids and proteins of C. elegans insulin-like genes and uses thereof

52
Priority: Apr 17, 1998Filed: Jan 9, 2003Published: Oct 2, 2003
Est. expiryApr 17, 2018(expired)· nominal 20-yr term from priority
Inventors:Stephen K. DobersteinAndrew Roy BuchmanKimberly Carr FergusonSheila Akiko HomburgerDarren Platt
A61K 38/00C07K 14/43545
52
PatentIndex Score
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Claims

Abstract

The present invention relates to C. elegans insulin-like genes and methods for identifying insulin-like genes. The methods provide nucleotide sequences of C. elegans insulin-like genes, amino acid sequences of their encoded proteins, and derivatives (e.g., fragments) and analogs thereof. The invention further relates to fragments (and derivatives and analogs thereof) of insulin-like proteins which comprise one or more domains of an insulin-like protein. Antibodies to an insulin-like protein, and derivatives and analogs thereof, are provided. Methods of production of an insulin-like protein (e.g., by recombinant means), and derivatives and analogs thereof, are provided. Further, methods to identify the biological function of a C. elegans insulin-like gene are provided, including various methods for the functional modification (e.g., overexpression, underexpression, mutation, knock-out) of one or more genes simultaneously. Still further, methods to identify a C. elegans gene which modifies the function of, and/or functions in a downstream pathway from, an insulin-like gene are provided.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A purified  C. elegans  insulin-like protein comprising an amino acid sequence of SEQ ID NO:198.  
     
     
         2 . A purified  C. elegans  insulin-like protein comprising amino acid numbers 19 to 103 of SEQ ID NO:198.  
     
     
         3 . A purified  C. elegans  insulin-like protein comprising an amino acid sequence of SEQ ID NO:199.  
     
     
         4 . A purified  C. elegans  insulin-like protein comprising amino acid numbers 19 to 72 of SEQ ID NO:199.  
     
     
         5 . A purified  C. elegans  insulin-like protein comprising an amino acid sequence of SEQ ID NO:200.  
     
     
         6 . A purified  C. elegans  insulin-like protein comprising amino acid numbers 17 to 110 of SEQ ID NO:200.  
     
     
         7 . A purified  C. elegans  insulin-like protein comprising amino acid sequence of SEQ ID NO:201.  
     
     
         8 . A purified  C. elegans  insulin-like protein comprising amino acid numbers 19 to 67 of SEQ ID NO:201.  
     
     
         9 . A purified  C. elegans  insulin-like protein comprising an amino acid sequence of SEQ ID NO:202.  
     
     
         10 . A purified  C. elegans  insulin-like protein comprising amino acid numbers 20 to 76 of SEQ ID NO:202.  
     
     
         11 . A purified  C. elegans  insulin-like protein comprising an amino acid sequence of SEQ ID NO:203.  
     
     
         12 . A purified  C. elegans  insulin-like protein comprising amino acid numbers 21 to 120 of SEQ ID NO:203.  
     
     
         13 . A purified  C. elegans  insulin-like protein comprising an amino acid sequence of SEQ ID NO:204.  
     
     
         14 . A purified  C. elegans  insulin-like protein comprising amino acid numbers 16 to 218 of SEQ ID NO:204.  
     
     
         15 . A purified  C. elegans  insulin-like protein comprising amino acid sequence of SEQ ID NO:205.  
     
     
         16 . A purified  C. elegans  insulin-like protein comprising amino acid numbers 29 to 107 of SEQ ID NO:205.  
     
     
         17 . A purified  C. elegans  insulin-like protein comprising amino acid sequence of SEQ ID NO:206.  
     
     
         18 . A purified  C. elegans  insulin-like protein comprising amino acid numbers 23 to 77 of SEQ ID NO:206.  
     
     
         19 . A purified derivative of the protein of any one of claims  1  through  18 , which derivative is capable of immunospecific binding to an anti-insulin-like protein antibody.  
     
     
         20 . A purified derivative of the protein of any one of claims  1  through  18 , which derivative displays one or more functional activities of the  C. elegans  insulin-like protein.  
     
     
         21 . A purified fragment of the protein of any one of claims  1  through  18 , which fragment displays one or more functional activities of the  C. elegans  insulin-like protein.  
     
     
         22 . A purified fragment of the protein of any one of claims  1  through  18 , comprising a domain of the  C. elegans  insulin-like protein selected from the group consisting of a B peptide domain and an A peptide domain.  
     
     
         23 . A purified molecule comprising the fragment of  claim 21 .  
     
     
         24 . A purified molecule comprising the fragment of  claim 22 .  
     
     
         25 . A chimeric protein comprising-the fragment of  claim 21 , consisting of at least 6 amino acids fused by a covalent bond to an amino acid sequence of a second protein, which second protein is not a  C. elegans  insulin-like protein.  
     
     
         26 . A chimeric protein comprising the fragment of  claim 22 , consisting of at least 6 amino acids fused by a covalent bond to an amino acid sequence of a second protein, which second protein is not a  C. elegans  insulin-like protein.  
     
     
         27 . The chimeric protein of  claim 25 , wherein the fragment of the  C. elegans  insulin-like protein is a fragment capable of immunospecific binding to an anti-insulin-like protein antibody.  
     
     
         28 . The chimeric protein of  claim 26 , wherein the fragment of the  C. elegans  insulin-like protein is a fragment capable of immunospecific binding to an anti-insulin-like protein antibody.  
     
     
         29 . A purified antibody or an antigen-binding derivative thereof capable of immunospecific binding to the protein of any one of claims  1  through  18  and not to an insulin-like protein of another species.  
     
     
         30 . The antibody of  claim 29  which is polyclonal.  
     
     
         31 . The antibody of  claim 29  which is monoclonal.  
     
     
         32 . An isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214 and SEQ ID NO:215.  
     
     
         33 . An isolated nucleic acid comprising a nucleotide sequence encoding an amino acid sequence selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206.  
     
     
         34 . The nucleic acid of  claim 32  or  claim 33  which is cDNA.  
     
     
         35 . The nucleic acid of  claim 32  or  claim 33  which is mRNA.  
     
     
         36 . An isolated nucleic acid which hybridizes under conditions of high stringency to a second nucleic acid consisting of said nucleotide sequence of the nucleic acid of  claim 32  or  claim 33 .  
     
     
         37 . The nucleic acid of  claim 36  which encodes a  C. elegans  insulin-like protein or a functional derivative thereof.  
     
     
         38 . An isolated nucleic acid comprising a nucleotide sequence encoding a functional derivative of an amino acid sequence selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206.  
     
     
         39 . An isolated nucleic acid comprising a nucleotide sequence that is antisense to a nucleotide sequence selected from the group consisting of SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214 and SEQ ID NO:215.  
     
     
         40 . An isolated nucleic acid comprising a nucleotide sequence that is antisense to a nucleotide sequence encoding an amino acid sequence selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206.  
     
     
         41 . A method of producing a  C. elegans  insulin-like protein comprising: 
 (a) growing a recombinant cell containing a nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214 and SEQ ID NO:215 such that the encoded  C. elegans  insulin-like protein is expressed by the cell; and    (b) recovering the expressed  C. elegans  insulin-like protein.    
     
     
         42 . A purified  C. elegans  insulin-like protein produced by the method of  claim 41 .  
     
     
         43 . A method of producing a  C. elegans  insulin-like protein comprising: 
 (a) growing a recombinant cell containing a nucleic acid comprising a nucleotide sequence encoding a  C. elegans  insulin-like protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206 such that the encoded  C. elegans  insulin-like protein is expressed by the cell; and    (b) recovering the expressed  C. elegans  insulin-like protein.    
     
     
         44 . A purified  C. elegans  insulin-like protein produced by the method of  claim 43 .  
     
     
         45 . A method of identifying a phenotype associated with mutation or abnormal expression of a  C. elegans  insulin-like protein comprising identifying the effect of a mutated or abnormally expressed  C. elegans  insulin-like gene which encodes a  C. elegans  insulin-like protein selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206 in a  C. elegans  animal.  
     
     
         46 . The method of  claim 45 , wherein the effect is determined by an assay selected from the group consisting of a dauer formation assay, a developmental assay, an energy metabolism assay, a growth rate assay and a reproductive capacity assay.  
     
     
         47 . A method of identifying a phenotype associated with mutation or abnormal expression of a  C. elegans  insulin-like protein comprising: 
 (a) mutating or abnormally expressing a  C. elegans  insulin-like gene which encodes a  C. elegans  insulin-like protein selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206 in a  C. elegans  animal; and    (b) identifying an effect of the gene mutated or abnormally expressed.    
     
     
         48 . The method of  claim 47 , wherein the effect is identified by an assay selected from the group consisting of a dauer formation assay, a developmental assay, an energy metabolism assay, a growth rate assay and a reproductive capacity assay.  
     
     
         49 . The method of any one of claims  45  through  48 , herein the gene is mutated or abnormally expressed using a technique selected from the group consisting of EMS chemical deletion mutagenesis, transposon insertion mutagenesis and double-stranded RNA interference.  
     
     
         50 . A recombinant cell containing a recombinant nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214 and SEQ ID NO:215.  
     
     
         51 . A vector comprising (a) a nucleotide sequence selected from the group consisting of SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214 and SEQ ID NO:215, and (b) an origin of replication.  
     
     
         52 . The vector of  claim 51  in which the nucleotide sequence is operably linked to a heterologous promoter.  
     
     
         53 . A purified  C. elegans  insulin-like protein encoded by a nucleic acid capable of hybridizing under conditions of high stringency to a nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214 and SEQ ID NO:215.  
     
     
         54 . A purified protein consisting of a B peptide domain defined by amino acid numbers 19-77 of SEQ ID NO:198 linked by one or more disulfide bonds to an A peptide domain defined by amino acid numbers 78-103 of SEQ ID NO:198.  
     
     
         55 . A purified protein consisting of a B peptide domain defined by amino acid numbers 19-49 of SEQ ID NO:199 linked by one or more disulfide bonds to an A peptide domain defined by amino acid numbers 50-72 of SEQ ID NO:199.  
     
     
         56 . A purified protein consisting of a B peptide domain defined by amino acid numbers 17-82 of SEQ ID NO:200 linked by one or more disulfide bonds to an A peptide domain defined by amino acid numbers 83-110 of SEQ ID NO:200.  
     
     
         57 . A purified protein consisting of a B peptide domain defined by amino acid numbers 19-45 of SEQ ID NO:201 linked by one or more disulfide bonds to an A peptide domain defined by amino acid numbers 46-67 of SEQ ID NO:201.  
     
     
         58 . A purified protein-consisting of a B peptide domain defined by amino acid numbers 21-51 of SEQ ID NO:202 linked by one or more disulfide bonds to an A peptide domain defined by amino acid numbers 52-76 of SEQ ID NO:202.  
     
     
         59 . A purified protein consisting of a B peptide domain defined by amino acid numbers 21-90 of SEQ ID NO:203 linked by one or more disulfide bonds to an A peptide domain defined by amino acid numbers 91-120 of SEQ ID NO:203.  
     
     
         60 . A purified protein consisting of a first B peptide domain defined by amino acid numbers 16-50 of SEQ ID NO:204 linked by one or more disulfide bonds to a first A peptide domain defined by amino acid numbers 51-89 of SEQ ID NO:204, a second B peptide domain defined by amino acid numbers 90-110 of SEQ ID NO:204 linked by one or more disulfide bonds to a second A peptide domain defined by amino acid numbers 111-153 of SEQ ID NO:204, and a third B peptide domain defined by amino acid numbers 154-174 of SEQ ID NO:204 linked by one or more disulfide bonds to a third A peptide domain defined by amino acid numbers 175-218 of SEQ ID NO:204.  
     
     
         61 . A purified protein consisting of a B peptide domain defined by amino acid numbers 29-66 of SEQ ID NO:205 linked by one or more disulfide bonds to an A peptide domain defined by amino acid numbers 67-107 of SEQ ID NO:205.  
     
     
         62 . A purified protein consisting of a B peptide domain defined by amino acid numbers 23-47 of SEQ ID NO:206 linked by one or more disulfide bonds to an A peptide domain defined by amino acid numbers 48-77 of SEQ ID NO:206.  
     
     
         63 . A method of identifying a gene-of-interest as capable of modifying a function of a  C. elegans  insulin-like gene comprising: 
 (a) constructing a first mutant nematode having a first mutation in the  C. elegans  insulin-like gene which encodes a  C. elegans  insulin-like protein selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206 and a second mutation in the gene-of-interest; and    (b) determining whether the phenotype displayed by the first mutant nematode is different from the phenotype of a second mutant nematode having said first mutation but not said second mutation,    in which the displaying of a phenotype by the first mutant nematode that is different from said second mutant nematode identifies the gene-of-interest as capable of modifying the function of the  C. elegans  insulin-like gene.    
     
     
         64 . The method of  claim 63 , wherein the first mutant nematode is produced using a technique selected from the group consisting of EMS chemical deletion mutagenesis, transposon insertion mutagenesis and double-stranded RNA interference.  
     
     
         65 . The method of  claim 63 , wherein the phenotype is selected from the group consisting of an altered body shape phenotype, an altered body size phenotype, an altered chemotaxis phenotype, an altered brood size phenotype, an altered egg-laying phenotype, an altered life span phenotype, an altered lipid accumulation phenotype, an altered locomotion phenotype, an altered organ morphogenesis phenotype, an altered thermotaxis phenotype, a dauer constitutive phenotype, a dauer defective phenotype, a lethal phenotype and a sterile phenotype.  
     
     
         66 . The method of  claim 65 , wherein the altered organ morphogenesis phenotype involves an organ selected from the group consisting of vulva, nervous system, gut and musculature.  
     
     
         67 . The method of  claim 65 , wherein a nematode having the altered body size phenotype is assayed for activity of a gene affecting body size selected from the group consisting of daf-4, sma-2 and sma-3.  
     
     
         68 . The method of  claim 63 , wherein the gene-of-interest is a homolog of an insulin signaling pathway gene from vertebrates.  
     
     
         69 . The method of  claim 63 , wherein the gene-of-interest is selected from the group consisting of daf-2, daf-16 and age-1.  
     
     
         70 . A  C. elegans  animal having a first mutation in a  C. elegans  insulin-like gene comprising a cDNA selected from the group consisting of SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214 and SEQ ID NO:215, and a second mutation in a different gene that is a homolog of an insulin signaling pathway gene from vertebrates.  
     
     
         71 . A method of studying a function of a  C. elegans  insulin-like gene comprising: 
 (a) mis-expressing a wild-type or mutant  C. elegans  insulin-like gene which encodes a  C. elegans  insulin-like protein selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206 in a transgenic nematode by driving expression with a homologous or heterologous promoter; and    (b) detecting a phenotype in said transgenic nematode, so as to study the function of the  C. elegans  insulin-like gene.    
     
     
         72 . The method of  claim 71 , wherein the heterologous promoter driving mis-expression is selected from the group consisting of an hsp 16-2 promoter, an hsp 16-41 promoter, a myo-2 promoter, an hlh-1 promoter and a mec-3 promoter.  
     
     
         73 . The method of  claim 71 , wherein said transgenic nematode mis-expressing the  C. elegans  insulin-like gene further has a mutation in daf-2.  
     
     
         74 . The method of  claim 71 , wherein said transgenic nematode mis-expressing the  C. elegans  insulin-like gene is assayed for a change in a phenotype selected from the group consisting of dauer formation and life span.  
     
     
         75 . A method of detecting the effect of expression of a  C. elegans  insulin-like gene which encodes a  C. elegans  insulin-like protein selected from. the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206 on an insulin signaling pathway comprising: 
 (a) mutating or abnormally expressing a wild-type  C. elegans  insulin-like gene in a nematode already having a mutation in the insulin signaling pathway that displays a phenotype-of-interest; and    (b) detecting the effect of step (a) on the phenotype-of-interest,    so as to detect the effect of expression of the  C. elegans  insulin-like gene.    
     
     
         76 . The method of  claim 75 , wherein the mutation in the insulin signaling pathway is in a gene selected from the group consisting of daf-2, daf-16 and age-1.  
     
     
         77 . A method of identifying a molecule that specifically binds to a ligand selected from the group consisting of (i) a  C. elegans  insulin-like protein selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206, (ii) a fragment of the  C. elegans  insulin-like protein comprising a domain of the protein, and (iii) a nucleic acid encoding the  C. elegans  insulin-like protein or fragment, the method comprising: 
 (a) contacting the ligand with a plurality of molecules under conditions conducive to binding between the ligand and the molecules; and    (b) identifying a molecule within the plurality that specifically binds to the ligand.    
     
     
         78 . The method of  claim 77 , wherein the domain of the  C. elegans  insulin-like protein is. selected from the group consisting from a signal peptide domain, a pre peptide domain, a pro peptide domain, a B peptide domain, a C peptide domain and an A peptide domain.  
     
     
         79 . A recombinant non-human animal in which a  C. elegans  insulin-like gene which encodes a  C. elegans  insulin-like protein selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206 has been deleted or inactivated by recombinant methods, or a progeny thereof containing the deleted or inactivated gene.  
     
     
         80 . The recombinant non-human animal of  claim 79  wherein the  C. elegans  insulin-like gene has been deleted or inactivated by a method selected from the group consisting of EMS chemical deletion mutagenesis, transposon insertion mutagenesis and double-stranded RNA interference.  
     
     
         81 . A recombinant non-human animal containing a  C. elegans  insulin-like transgene which encodes a  C. elegans  insulin-like protein selected from the group consisting of SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205 and SEQ ID NO:206.  
     
     
         82 . The recombinant non-human animal of  claim 81 , wherein the  C. elegans  insulin-like transgene is under the control of a promoter that is not the native promoter of the transgene.  
     
     
         83 . A purified protein encoded by a first nucleic acid which hybridizes under conditions of high stringency to a second nucleic acid, which second nucleic acid comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:29, SEQ ID NO:29, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:163, SEQ ID NO:164, SEQ ID NO:165, SEQ ID NO:213, SEQ ID NO:214, and SEQ ID NO:215, wherein the protein is characterized as lacking a cleavable C peptide and as having the same number and relative spacing of Cys residues as found in vertebrate insulin-like proteins.  
     
     
         84 . A purified protein encoded by a first nucleic acid which hybridizes under conditions of high stringency to a second nucleic acid, which second nucleic acid comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32 and SEQ ID NO:33, SEQ ID NO:36, SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211 and SEQ ID NO:212 wherein the protein is characterized as (a) lacking a cleavable C peptide separating the B and A chains, (b) lacking an intra-chain disulfide bond in the A domain, and (c) having an extra pair of Cys residues relative to vertebrate insulin-like proteins.  
     
     
         85 . The protein of  claim 84  or  claim 85 , wherein the B and A chain domains of the protein are not proteolytically cleaved into separate chains.  
     
     
         86 . A method of identifying a molecule that alters the expression level of a  C. elegans  insulin-like gene corresponding to a cDNA selected from the group consisting of SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214 and SEQ ID NO:215, which method comprises: 
 (a) contacting a transgenic nematode with one or more molecules, said transgenic nematode having a transgene comprising a promoter or enhancer region of genomic DNA from 1 base to 6 kilobases upstream of the start codon of the cDNA operably linked to a reporter gene; and    (b) determining whether the level of expression of the reporter gene is altered relative to the level of expression of the reporter gene in the absence of the one or more molecules.    
     
     
         87 . A method of identifying a molecule that binds to a promoter or enhancer of a  C. elegans  insulin-like gene corresponding to a cDNA selected from the group consisting of SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214 and SEQ ID NO:215, which method comprises: 
 (a) contacting a transgenic nematode with one or more molecules, said transgenic nematode having a transgene comprising a promoter or enhancer region of genomic DNA from 1 base to 6 kilobases upstream of the start codon of the cDNA operably linked to a reporter gene;    (b) determining whether the level of expression of the reporter gene is altered relative to the level of expression of the reporter gene in the absence of the one or more molecules;    (c) contacting the one or more molecules with the promoter or enhancer region of genomic DNA; and    (d) identifying the molecule contacted in step (c) that binds to the promoter or enhancer.    
     
     
         88 . The method of  claim 86  or  claim 87 , wherein the reporter gene encodes green fluorescent protein.

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