基于错配识别蛋白MutS的高特异性的单碱基错配核酸检测方法  

High specificity single base mismatch nucleic acid detection based on MutS

作　　者：袁莹[1] 张斌[2] 李硕[1] 关一夫[1] YUAN Ying;ZHANG Bin;LI Shuo;GUAN Yi-fu(Department of Biochemistry and Molecular Biology,China Medical University,Shenyang 110122;Department of Oral-maxillofacial Surgery,School and Hospital of Stomatology,China Medical University,Shenyang 110001 China)

机构地区：[1]中国医科大学生命科学学院生物化学与分子生物学教研室,辽宁沈阳110122 [2]中国医科大学附属口腔医学院口腔颌面外科教研室,辽宁沈阳110001

出　　处：《解剖科学进展》2020年第4期450-453,共4页Progress of Anatomical Sciences

基　　金：国家自然科学基金(31670821);辽宁省第三批新冠肺炎疫情防控应急科研攻关计划(2020JH2/10300024)。

摘　　要：目的建立一种基于错配识别蛋白的高特异性核酸检测方法。方法利用蛋白质体外表达和纯化技术获取MutS,并对MutS的错配识别特性进行测定;利用MutS-RCA检测体系对2个碱基错配的不同DNA片段进行检测。结果 MutS蛋白能够有效区分开互补双链和非互补双链DNA,能够稳定结合在非互补双链DNA上,基于MutS的高特异性的单碱基错配核酸检测方法取得了良好的特异性。结论 MutS-RCA检测体系提高了单碱基错配核酸检测的特异性。Objective To establish a high specificity nucleic acid detection method based on mismatch recognition protein MutS. Methods MutS was obtained by protein expression and purification in vitro, and the recognition characteristics of MutS were determined. Different DNA fragments were detected by MutS-RCA detection. Results MutS protein distinguished effectively the complementary double stranded DNA from non-complementary double stranded DNA, and stably binded to non-complementary double stranded DNA. A good specificity was obtained by single base mismatch nucleic acid detection based on MutS-RCA detection system. Conclusion MutS-RCA detection system would provide powerful technical support for reducing the non-specific detection caused by single base mismatch.

关 键 词：错配识别蛋白 滚环扩增 单碱基错配 

分 类 号：R446.1[医药卫生—诊断学]