PCR-DGGE扫查胰高糖素受体基因的实验条件研究  

Study of the Experimental Conditions of PCR-DGGE in Scanning the Glucagon Receptor Gene

作　　者：王玉明[1] 段勇[1] 宋滇平[1] 赵淮[1] 杨慧英[1] 刘华[1] 

机构地区：[1]昆明医学院第一附属医院,昆明650032

出　　处：《昆明医学院学报》1999年第4期1-4,共4页Journal of Kunming Medical College

基　　金：云南省应用基础研究基金! ( 96C13 5M);国家人事部非教育系统留学回国人员科技活动择优资助经费资助

摘　　要：为确定聚合酶链反应—变性梯度凝胶电泳(PCR-DGGE)扫查胰高糖素受体(GCG-R)基因的主要实验条件,对PCR-DGGE扫查GCG-R基因各个外显子时其最适PCR反应条件、变性剂浓度范围和电泳时间进行了研究.结果表明,当引物富含GC碱基时其最适PCR反应条件与扩增无GC-clamp之DAN片段略有不同,不同碱基组成及不同片段长度的DNA分子其在DGGE系统中所需的变性剂浓度范围和电泳时间不尽相同.提示合理的变性剂浓度范围和电泳时间是DGGE实验的关键因素.To probe into the main experimental conditions in polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) scanning of the glucagon receptor (GCG-R) gene, the optimum PCR system, denaturant concentration range and electrophoresis time were studied. The results showed that there were some difference in PCR systems between the amplification of DNA fragments with and without a GC-rich segment. The denaturant concentration range and electrophoresis time in DGGE depended upon the melting behavior of the test DNA molecules based on their nucleotide sequences and length. It was suggested that the denaturant concentration range and electrophoresis time may be the key factors for DGGE experiment.

关 键 词：胰高糖素受体 基因 糖尿病 PCR-DGGE 

分 类 号：R587.104[医药卫生—内分泌]