不对称PCR-SSCP在基因突变检测中的应用  

Application of asymmetric PCR-SSCP in gene mutation detecting

作　　者：张小辉[1] 许尚忠[2] 高雪[2] 张路培[2] 任红艳[2] 陈金宝[2] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]中国农业科学院畜牧研究所,北京100094

出　　处：《西北农林科技大学学报（自然科学版）》2007年第6期15-18,23,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"863"计划项目(2002AA242011)

摘　　要：为了研究不对称PCR-SSCP在基因突变检测中的准确性,以鲁西黄牛和荷斯坦奶牛为研究对象,用不对称PCR-SSCP分析技术分析了牛SMAD4基因3′端非翻译区的碱基突变情况,比较了不对称PCR-SSCP和传统PCR-SSCP分析的优缺点。结果表明,鲁西黄牛和荷斯坦奶牛SMAD4基因3′端非翻译区有1个T碱基插入突变位点和1个G→A突变位点,其中G→A突变产生了1个HhaⅠ酶切位点;不对称PCR-SSCP和HhaⅠ酶切分析116头鲁西黄牛和75头荷斯坦奶牛群体G→A突变位点的频率完全一致,说明不对称PCR-SSCP不仅可以用于基因突变的检测,而且具有较高的准确性;不对称PCR-SSCP较传统PCR-SSCP的条带少且带型清晰、稳定性较高。以上结果表明,不对称PCR-SSCP可以用于基因突变的检测。The advantage and disadvantage of asymmetric PCR-SSCP and traditional PCR-SSCP were compared to analyze the character and accurate rate in detecting mutation in this study. The mutations in 3r UTR region of SMAD4 gene were analyzed by asymmetric PCR-SSCP in Luxi cattle and Holstein cow and one insert T mutation and one G→A mutation were found in this region. The G→A mutation created a Hha I enzyme digestion position and the frequency was studied by asymmetric PCR-SSCP and enzyme digestion in 116 Luxi cattle and 75 Holstein cows and got the same results. The asymmetric PCR-SSCP had a few bands,which were clearer and stabler than traditional PCR-SSCP. This indicated the asymmetric PCR- SSCP was suitable for mutations detecting.

关 键 词：不对称PCR-SSCP 基因突变 SMAD4 

分 类 号：S813.3[农业科学—畜牧学]