采用PCR方法检测Fmr1基因敲除小鼠的基因型  

GENOTYPE ANALYSIS OF FMR1 GENE KNOCKOUT MICE WITH POLYMERASE CHAIN REACTION

作　　者：邢州[1] 孙卫文[1] 黄越玲[2] 易咏红[1] 戴丽军[2] 陈盛强[1] 李敏雄[1] 

机构地区：[1]广州医学院第二附属医院,广东广州510260 [2]广州医学院动物实验中心,广东广州510182

出　　处：《现代医院》2009年第5期12-14,共3页Modern Hospitals

基　　金：广东省自然科学基金项目(编号:8012011);广东省科技计划项目(编号:2005B60302004;2008B030301371);广东省中医药管理局建设中医药强省科研基金项目(编号:2008184);广州医学院留学归国启动项目(编号:0707085);广州市中医药中西医结合科研立项项目(编号:2008A52)

摘　　要：目的为Fmr1基因敲除小鼠探索快速、简单的基因型PCR检测方法。方法设计两对引物扩增野生型Fmr1基因和Fmr1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western blot方法比较。结果野生型仅在800bp处有一条条带,突变纯合子仅在500bp处有一条条带,杂合子则在500bp和800bp处出现两条条带。用PCR方法获得的Fmr1基因分析结果与经典的western blot方法获得的结果完全一致。结论用PCR方法分析Fmr1基因敲除鼠的基因型具有快速、简单、廉价和适用的特点。Objective To explore a simple and quick polymerase chain reaction （PCR） method for the genotyping of Fmrl knockout mice. Methods 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 468 bp band was found in wild - type mice,a 800 bp band in homozygous mutated Fmrl 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：聚合酶链反应 基因型 WESTERN BLOT 基因敲除 FMR1 

分 类 号：R379.4[医药卫生—病原生物学] R363.1[医药卫生—基础医学]