[Detection of single-nucleotide polymorphisms in insulin receptor substrate-2 gene 3'-untranslated region by denaturing high-performance liquid chromatography].
OBJECTIVE
To explore the use of denaturing high-performance liquid chromatography (DHPLC) in detecting single-nucleotide polymorphisms(SNPs) of insulin receptor substrate-2(IRS-2) gene 3'-untranslated region (3'-UTR).


METHODS
We detected the SNPs and mutation of IRS-2 gene 3'-UTR sequence, with polymerase chain reaction (PCR), DHPLC, single-strand conformation polymorphism (SSCP), and DNA sequence analysis respectively in 30 Type 2 diabetic subjects and 30 healthy controls.


RESULTS
The SNPs of IRS-2 gene 3'-UTR in 4 patients with Type 2 diabetes mellitus were detected by PCR-DHPLC and were identified by DNA sequencing.


CONCLUSIONS
DHPLC is a rapid and automated technology, which is simpler and more accurate for detecting longer fragment of DNA than SSCP. The present method lays a foundation for further studying the relationship between the mutation of IRS-2 gene 3'-UTR and Type 2 diabetes mellitus.
Zeng
Wei-min
WM
Department of Biochemistry, Xiangya School of Medicine, Central South University, Changsha 410078, China.
Chen
Shu-hua
SH
Xie
Ping
P
D020413:3' Untranslated Regions / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003924:Diabetes Mellitus, Type 2 / Q000235:genetics*; D006801:Humans; D055504:Insulin Receptor Substrate Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009691:Nucleic Acid Denaturation; D010750:Phosphoproteins / Q000235:genetics*; D017354:Point Mutation; D020641:Polymorphism, Single Nucleotide; D018807:Polymorphism, Single-Stranded Conformational
D020413:3' Untranslated Regions; C526297:IRS2 protein, human; D055504:Insulin Receptor Substrate Proteins; D047908:Intracellular Signaling Peptides and Proteins; D010750:Phosphoproteins