PCR方法在HSF1基因敲除小鼠基因型分析中的应用  

Genotype Analysis of HSF1 Gene Knockout Mice with Polymerase Chain Reaction

作　　者：陈广文[1] 刘喜玲[2] 肖献忠[1] 

机构地区：[1]中南大学湘雅医学院病理生理学教研室,长沙410078 [2]卫生部肝胆肠外科中心医学工程研究室

出　　处：《中国实验动物学报》2002年第2期73-76,共4页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金"α- B晶状体蛋白保护心肌细胞缺氧损伤研究"(3 9770 3 0 7)

摘　　要：目的　为HSF1基因敲除鼠探索快速、简单的基因型PCR检测方法。方法　设计两对引物扩增野生型HSF1基因和HSF1缺陷突变基因的DNA片段 ,用PCR仪梯度方案测试最佳退火温度 ,并将所得基因型结果与经典的Southernblot方法比较。结果　野生型仅在 5 6 2bp处有一条条带 ,突变纯合子仅在 377bp处有一条条带 ,杂合子则在377bp和 5 6 2bp处出现两条条带。用PCR方法获得的HSF1基因分析结果与经典的Southernblot方法获得的结果完全一致。结论　用PCR方法分析HSF1基因敲除鼠的基因型具有快速、简单。Objective This study was to explore a simple and quick polymerase chain reaction (PCR) method for the genotyping of heat shock factor 1(HSF1) knockout mice. Method\ Two pairs of primers were designed to amplify genomic DNA fragment of wild-type HSF1 gene and the same region on HSF1 targeting vector respectively. A gradient PCR strategy was used to test the best annealing temperature. Results\ A 562 bp band was found in wild-type HSF1 mice, a 377 bp band in homozygous mutated HSF1 mice and both bands in heterozygous mice. The genotyping results were completely coincided with those from typical Southern blot. Conclusion \ PCR is a simple, fast and practical method for the genotyping of HSF1 gene knockout mice.

关 键 词：PCR方法 HSF1基因敲除小鼠 基因型分析 应用 

分 类 号：R－332[医药卫生] R394－33