PCR-ACRS方法筛查CYP21基因P459H的突变位点  

Screening of CYP21 gene P459H mutation by PCR-ACRS

作　　者：王甲莉[1] 蒋玲[1] 宋璐璐[1] 王慧[2] 

机构地区：[1]山东大学齐鲁医院内分泌科,山东济南250012 [2]北京中日友好医院内分泌科,北京100029

出　　处：《中国病理生理杂志》2009年第1期168-171,共4页Chinese Journal of Pathophysiology

基　　金：山东省科技攻关基金资助项目(No.2006GG2302023);山东省自然科学基金资助项目(No.Y2007C105)

摘　　要：目的:研究21-羟化酶基因(CYP21)第10外显子P459H突变(CCC→CAC)在正常人群中的发生率,分析P459H是点突变还是基因多态。方法:设计特异性引物扩增CYP21基因3-10外显子,PstⅠ限制性内切酶消化验证PCR产物为特异性扩增,在此基础上,应用PCR结合扩增引进限制性酶切位点方法(PCR-ACRS)扩增首次PCR产物,FspⅠ限制性内切酶酶切后用10%聚丙烯酰胺凝胶电泳筛查第10外显子P459H的发生率。结果:100例正常人CYP21基因第10外显子密码子459均为CCC,即均未出现P459H变异。结论:CYP21基因P459H为点突变,排除基因多态的可能;另外,PCR-ACRS是一种快速、可靠的检测基因点突变的有效途径。AIM： To investigate the frequency of P459H （CCC→CAC） in exon 10 of CYP21 gene among normal population. METHODS： The exons 3 -10 of CYP21 gene were amplified with polymerase chain reaction （PCR）. The PCR round products were digested by restriction enzyme Pst Ⅰ to confirm that CYP21 gene was specifically amplified. PCR - based amplification - created restriction site （PCR - ACRS） was performed using the first round PCR products as template. After the second PCR products were digested by restriction enzyme Fsp Ⅰ, 10% polyacrylamide gel electrophoresis was used to screen the frequency of P459H in exon 10. RESULTS ： The codon 459 in exon 10 of CYP21 gene was all CCC among 100 normal cases tested. CONCLUSION： P459H （CCC →CAC） of CYP21 gene might be a novel point mutation causing CAH. Furthermore, PCR - ACRS was a fast and safe method for gene mutation screening.

关 键 词：基因 CYP21 点突变 肾上腺增生 先天性 

分 类 号：R586[医药卫生—内分泌]