Isolated nucleic acids and polypeptides associated with glucose homeostasis disorders and method of detecting the same

Abstract

An isolated and substantially pure nucleic acid sequence located between D20S119 and D20S178 on human chromosome 20q13, the nucleic acid sequence including: a nucleic acid sequence coding for a glucose transporting protein and having the sequence shown in SEQ ID NO: 1; or a nucleic acid sequence having at least 70% sequence identity with the nucleic acid sequence shown in SEQ ID NO: 1. The disclosed nucleic acid sequences map to a locus associated with human Type II diabetes mellitus and, therefore, therapeutic and diagnostic screening methods, which accommodate naturally and artificially occurring polymorphisms, are also disclosed.

1 - 8 . (canceled) 
     
     
         9 . A method of facilitating a diagnosis of susceptibility to a glucose homeostasis disorder, the method comprising:
 (a) obtaining a biological sample from a subject;   (b) determining a sequence of a target nucleic acid sequence from the biological sample, wherein the target nucleic acid sequence is a glucose transporter gene located on chromosome 20q13 between D20S119 and D20S178;   and   (c) determining a sequence variation between a wild type nucleic acid sequence and the target nucleic acid sequence, the presence of a variation between the wild type nucleic acid sequence and the target nucleic acid sequence indicating susceptibility of the subject to a glucose homeostasis-related disorder.   
     
     
         10 . The method of  claim 9 , wherein the target nucleic acid sequence is a nucleic acid sequence which hybridizes to and is at least 70% complementary to the glucose transporter gene of SEQ ID NO: 1 and the wild type nucleic acid sequence is SEQ ID NO: 1. 
     
     
         11 . (canceled) 
     
     
         12 . The method of  claim 9 , wherein the disorder affecting glucose homeostasis is human Type II diabetes mellitus. 
     
     
         13 . The method of  claim 9 , wherein the biological sample is a tissue sample or a blood sample. 
     
     
         14 . The method of  claim 9 , wherein the determining the sequence is performed using a dideoxy sequencing method. 
     
     
         15 . The method of  claim 9 , wherein the target nucleic acid sequence is amplified prior to the determining the sequence. 
     
     
         16 - 20 . (canceled) 
     
     
         21 . A method of identifying a subject having a polymorphism of the nucleic acid sequence shown in SEQ ID NO: 1, the method comprising:
 (a) sequencing a target nucleic acid of a sample from a subject by dideoxy sequencing; and   b) identifying a polymorphism by comparing the nucleic acid sequence of the target nucleic acid sequence to the DNA sequence shown in SEQ ID NO: 1.   
     
     
         22 . The method of  claim 21 , wherein the sequencing is performed following amplification of the target nucleic acid. 
     
     
         23 - 66 . (canceled) 
     
     
         67 . A method of facilitating a diagnosis of susceptibility to a glucose homeostasis disorder by determining the presence of a genetic variation in or near a glucose transporter gene, comprising:
 a) obtaining a biological sample from a subject; and   b) performing a mutation analysis of genomic DNA and/or RNA from the biological sample,   
       wherein performing the mutation analysis determines whether there is a genetic variation in or near the glucose transporter gene. 
     
     
         68 . The method of claim  23 , wherein the subject is a human. 
     
     
         69 . The method of claim  23 , wherein the disorder affecting glucose homeostasis is human Type II diabetes mellitus. 
     
     
         70 . The method of claim  23 , wherein the biological sample is a tissue sample or a blood sample. 
     
     
         71 . The method of claim  23 , wherein the genetic variation is one or more of a polymorphism, a single nucleotide polymorphism (SNP), a chromosomal translocation, a chromosomal inversion, or a repeat expansion. 
     
     
         72 . The method of claim  23 , wherein the mutation analysis is performed by a technique selected from the group consisting of: a single-strand conformation polymorphism (SSCP) analysis, a SSCP/heteroduplex analysis, an enzyme mismatch cleavage, an allele-specific hybridization, a restriction analysis of the genomic DNA and/or RNA, and a direct sequence analysis of the genomic DNA and/or RNA. 
     
     
         73 . The method of claim  28 , wherein the mutation analysis is performed by the direct sequence analysis and the genomic DNA and/or RNA is amplified prior to the direct sequence analysis. 
     
     
         74 . The method of claim  28 , wherein the direct sequence analysis is performed using one or more oligonucleotide primers having a design based on an intronic sequence flanking an exon of the glucose transporter gene shown in Table 2. 
     
     
         75 . The method of claim  28 , wherein the direct sequence analysis is performed using one or more oligonucleotide primers having a design based on an exon of the glucose transporter gene of SEQ ID NO: 1. 
     
     
         76 . The method of claim  28 , wherein the genetic variation is a single nucleotide polymorphism (SNP) and the mutation analysis is the allele-specific hybridization, the direct sequence analysis of the genomic DNA and/or RNA, or the restriction analysis of the genomic DNA and/or RNA. 
     
     
         77 . The method of claim  28 , wherein the genetic variation is a repeat expansion and the mutation analysis comprises PCR of the genomic DNA and/or RNA with one or more oligonucleotide primers flanking the repeat.