碱基错配长PCR引物法检测CCNH基因rs3093816位点的多态性  

Development of mismatched long PCR primers for analysis of rs3093816 polymorphism in the CCNH gene

作　　者：常伟 丁明翠 王威 CHANG Wei;DING Mingcui;WANG Wei(Center for Disease Control,General Hospital of Pingmei Shenma Medical Group,Pingdingshan 467002;Department of Occupational Health and Occupational Disease,College of Public Health,Zhengzhou University,Zhengzhou 450001,Henan,China)

机构地区：[1]平煤神马医疗集团总医院疾控中心,河南平顶山467002 [2]郑州大学公共卫生学院劳动卫生与职业病学教研室,河南郑州450001

出　　处：《癌变．畸变．突变》2022年第2期129-133,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：郑州大学高层次人才启动基金(32340132);平煤神马医疗集团总医院2018年科技计划项目。

摘　　要：目的:细胞周期蛋白H(CCNH)基因rs3093816位点的多态性与许多癌症的发生风险升高有关,但由于此位点缺少合适的限制性内切酶位点,使聚合酶链式反应-限制性片段长度多态性技术(PCR-RFLP)的使用受到限制。因此,本研究拟建立碱基错配长PCR引物法用于检测CCNH基因rs3093816位点。方法:通过创造性酶切位点PCR-RFLP方法在CCNH扩增产物中引入新的酶切位点,然后利用Cvi Q I酶区分PCR产物。本研究同时设计了碱基错配长PCR引物和碱基错配相对短PCR引物,比较分析碱基错配长PCR引物是否更简便经济。结果:与短引物法相比,长引物占扩增总片段的比例越大,酶切产物越容易区分,凝胶电泳需要的时间越短。结论:成功建立了错配长PCR引物法检测CCNH基因rs3093816位点。OBJECTIVE:It has been demonstrated that the single nucleotide polymorphism(SNP)in the rs3093816 site of the cyclin H(CCNH)gene was associated with an increased cancer risk.There is,however,a limitation to utilizing PCR-RFLP due to the lack of proper restriction enzyme sites in rs3093816.Therefore,an appropriate method for identifying the polymorphism was developed in this study.METHODS:A new restriction site was introduced into the CCNH amplification products using a created-restriction site PCR(CRS PCR)which allowed the enzymes CviQ I in distinguishing the PCR products.In addition,long primers were compared with relatively short primers to investigate whether the long primers are more economical and modest.RESULTS:Compared with the short primers,the larger the proportion of the long primers to the total amplified fragment,the easier to distinguish the digestion products,and the shorter the time required for electrophoresis.CONCLUSION:A mismatched long PCR primers method was successfully developed to detect the rs3093816 polymorphism in the CCNH gene.

关 键 词：细胞周期蛋白H 创造性酶切位点 单核苷酸多态性 聚合酶链式反应-限制性片段长度多态性技术 

分 类 号：R394-3[医药卫生—医学遗传学]