三种常见CD36基因突变体标准质粒的构建  

Construction of three common CD36 gene mutant standard plasmids

作　　者：陈尚良 刘正婷 黄佳丽 何海洪 黎绍昌 李立浩 CHEN Shangliang;LIU Zhengting;HUANG Jiali;HE Haihong;LI Shaochang;LI Lihao(Shenzhen Hospital,Southern Medical University,Shenzhen,Guangdong 518000,China)

机构地区：[1]南方医科大学深圳医院,广东深圳518000

出　　处：《检验医学与临床》2020年第12期1659-1661,1665,共4页Laboratory Medicine and Clinic

基　　金：广东省深圳市科技计划(JCYJ20170307143728891);广东省深圳市宝安区科技计划(2016CX306)。

摘　　要：目的构建3种常见的CD36基因突变质粒,为建立高分辨率熔解曲线方法提供突变标准品。方法选取广州人群常见的CD36基因突变位点A1237C、C268T及329-330delAC作为研究位点,采用重叠延伸PCR方法及TA克隆技术构建含有上述突变位点的野生型和突变型质粒,并且对其进行测序验证。结果DNA测序结果证实构建的3种重组质粒分别含有CD36基因A1237C、C268T、329-330delAC突变。结论成功构建CD36基因A1237C、C268T、329-330delAC突变体标准质粒,为后续建立高分辨率熔解曲线方法筛查CD36基因多态性奠定物质基础。Objective Three common CD36 gene mutation plasmids were constructed to provide mutation standards for establishing a high-resolution melting curve method.Methods The CD36 gene mutation sites A1237C,C268T and 329-330delAC,which were common in Guangzhou population,were selected as research sites.The overlapping extension PCR method and TA cloning technology were used to construct the wild-type and mutant-type plasmids containing the above-mentioned mutation sites,and they were verified by sequencing.Results DNA sequencing results confirmed that the three recombinant plasmids contained CD36 gene A1237C,C268T,and 329-330delAC mutations,respectively.Conclusion The CD36 gene A1237C,C268T,329-330delAC mutant standard plasmids were successfully constructed,which laid a material foundation for the subsequent development of a high-resolution melting curve method for screening CD36 gene polymorphisms.

关 键 词：CD36基因 基因突变 重叠延伸PCR TA克隆 

分 类 号：Q343.13[生物学—遗传学]