碱基错配长PCR引物法检测CCNH基因rs3093816位点的多态性

doi:10.3969/j.issn.1004-616x.2022.02.010

癌变·畸变·突变 ›› 2022, Vol. 34 ›› Issue (2): 129-133.doi: 10.3969/j.issn.1004-616x.2022.02.010

• 技术与方法 • 上一篇

碱基错配长PCR引物法检测CCNH基因rs3093816位点的多态性

常伟¹, 丁明翠², 王威²

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

收稿日期:2021-09-06 修回日期:2022-02-22 发布日期:2022-04-07
通讯作者: 王威,E-mail:ww375@126.com E-mail:ww375@126.com
作者简介:常伟,E-mail:2540664578@qq.com。
基金资助:
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

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

CHANG Wei¹, DING Mingcui², WANG Wei²

1. Center for Disease Control, General Hospital of Pingmei Shenma Medical Group, Pingdingshan 467002;
2. Department of Occupational Health and Occupational Disease, College of Public Health, Zhengzhou University, Zhengzhou 450001, Henan, China

Received:2021-09-06 Revised:2022-02-22 Published:2022-04-07

摘要/Abstract

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CviQ 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。

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

Abstract: 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 (CRSPCR) 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.

Key words: cyclin H, created restriction site, single nucleotide polymorphism, PCR-RFLP

中图分类号:

R394-3

常伟, 丁明翠, 王威. 碱基错配长PCR引物法检测CCNH基因rs3093816位点的多态性[J]. 癌变·畸变·突变, 2022, 34(2): 129-133.

CHANG Wei, DING Mingcui, WANG Wei. Development of mismatched long PCR primers for analysis of rs3093816 polymorphism in the CCNH gene[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(2): 129-133.

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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

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