Assay for the detection of factors that modulate the expression of inagp

Abstract

A reporter construct contains mammalian INGAP 5′-regulatory region or a fragment thereof, a minimal promoter element from mammalian INGAP or a heterologous promoter, and a reporter gene. The reporter construct can be used to screen for agents which alone or in combination up-regulate or down-regulate reporter gene expression. Alternatively, the reporter construct can be used to screen for agents that bind to the hamster INGAP 5′-regulatory region or a fragment thereof.

1 . A method for identifying agents which modulate INGAP expression, comprising:
 contacting a host cell comprising a reporter construct having at least one INGAP regulatory region and a region encoding a detectable product with a test agent;   determining expression of the detectable product in the cell; and identifying the test agent as a modulator of INGAP expression if the test agent modulates expression of the detectable product in the cell.   
     
     
         2 . The method of  claim 1  wherein the regulatory region nucleotide sequence comprises of one or more regions chosen from nucleotides from the group consisting of SEQ ID NO: 1, 2, 23, 32, 35, 37, 28, 24, 25, 26, 27, 29, 30, 31, 33, 34, 38, and 36. 
     
     
         3 . The method of  claim 6  wherein the regulatory sequence comprises nucleotide sequences 1-3137 of SEQ ID NO: 2 
     
     
         4 . The method of  claim 2  wherein the reporter construct, further comprises: a promoter element interposed between the regulatory region nucleotide sequence and the nucleotide sequence encoding the detectable product. 
     
     
         5 . The method of  claim 4  wherein the promoter element is selected from SEQ ID NO: 2. 
     
     
         6 . An in vitro method for identifying agents which modulate INGAP expression, comprising: a. contacting a reporter construct having at least one INGAP regulatory region and a region encoding a detectable product with a test substance under conditions sufficient for transcription and translation of said nucleotide sequence; determining expression of the detectable protein or nucleic acid product; and identifying the test substance as a modulator of INGAP expression if the test substance modulates expression of the detectable product. 
     
     
         7 . The method of  claim 6  wherein the regulatory region nucleotide sequence comprises of one or more regions chosen from nucleotides from the group consisting of SEQ ID NO: 1, 2, 23, 32, 35, 37, 28, 24, 25, 26, 27, 29, 30, 31, 33, 34, 38, and 36. 
     
     
         8 . The method of  claim 7  wherein the regulatory sequence comprises nucleotide sequences 1-3137 of SEQ ID NO: 2 
     
     
         9 . The method of  claim 6  wherein the reporter construct, further comprises: a promoter element interposed between the regulatory region nucleotide sequence and the nucleotide sequence encoding the detectable product. 
     
     
         10 . The reporter construct of  claim 9  wherein the promoter element is selected from SEQ ID NO: 2. 
     
     
         11 . An in vitro method for identifying agents which modulate INGAP expression, comprising: contacting the SEQ ID NOS 1, 2, 23, 32, 35, 37, 28, 24, 25, 26, 27, 29, 30, 31, 33, 34, 38, and 36 or fragments thereof with a test agent; determining binding of the test agent to the nucleic acid; and identifying the test agent as a potential modulator of INGAP expression if the test agent binds to the nucleic acid. 
     
     
         12 . The method of  claim 11  wherein the sequence comprises nucleotide sequences 1-3137 of SEQ ID NO: 2 
     
     
         13 . A method for inducing INGAP expression in a mammal in need thereof, comprising administering to the mammal an effective amount of a factor that stimulates INGAP expression in the said mammal. 
     
     
         14 . The method of  claim 13  wherein the factor that stimulates INGAP expression was identified by the methods of  claim 1 . 
     
     
         15 . The method of  claim 14  wherein the factor that stimulates INGAP expression was identified by the methods of  claim 11 . 
     
     
         16 . The method of  claim 13  wherein the factor that stimulates INGAP expression is selected from hLIF or PMA