应用mAPLP方法对湖南地区苗族人群进行mtDNA多态性研究  

mAPLP for Detection of mtDNA Polymorphism in Hunan Miao Population of China

作　　者：周海燕[1] 邹永华[1] 陈勇[1] 张蕊 倪斌[1] 

机构地区：[1]湖南省计划生育研究所省现代优生与技术重点实验室,长沙410002 [2]黑龙江省计划生育科学研究所,哈尔滨150020

出　　处：《国际遗传学杂志》2007年第6期409-412,共4页International Journal of Genetics

基　　金：湖南省科技项目(06SK3108)

摘　　要：目的应用多重扩增产物片段长度多态性分析方法(multiplex-amplified product-length polymorphism,mAPLP)建立线粒体DNA编码区单核苷酸多态性(single nucleotide polymorphism,SNP)快速、准确、有效的分型方法。方法根据线粒体DNA SNP位点(11969A、10398A、10400T、12811C、4580A、14979C)设计两条不同的正向或反向引物(使PCR扩增片段长度不同)和一条共用的反向或正向引物,运用多重PCR方法扩增,扩增产物经8%聚丙烯酰胺凝胶电泳分析。结果SNP基因座为片段大小不同的单一谱带,在湖南地区40名无血缘关系苗族个体中,6个SNP基因座的分布频率分别为0.025/0.975、0.200/0.800、0.200/0.800、0.250/0.750、1.000/0.000、1.000/0.000、0.025/0.975。共检出7种单体型,单体型分布频率为0.025、0.050、0.125、0.175、0.025、0.050、0.550。结论此方法是一种简单、快速、准确、有效的SNP分型方法,对建立mtDNA编码区SNP数据库,研究人类群体遗传学,进行法医学个体识别等具有很大的应用价值。Objective To develop a multiplex-amplified product-length polymorphism for single nucleotide polymorphism （SNP） loci typing in mitochondrial DNA coding regions. Methods We chose six mitochondrial DNA SNP loci and designed specific forwardor reverse primer with base different in size and a common reverse or forward primer. Results The genotyping of SNP was determined by the two allele specific fragments different in size after polyacrylamide gel and silv6r staining. In Chinese Hunan Miao population, the allelic frequencies of 11969A , 10398A, 10400T, 12811C ,4580A , 14979C were 0. 025/0.975,0.2/0.8,0.2/ 0. 8,0. 25/0. 75,1. 00/0,1. 00/0,0. 025/0. 975, respectively. A total of 7 different haplotypes was identified and Haplotype frequencies were 0. 025,0.05,0. 125,0. 175,0. 025 ,＇0.05,0.55 respectively. Conclusion mAPLP was a simple, rapid, accurate and efficient method for SNP typing, which will be very useful for SNP in the database establishing of mitochondrial DNA coding regions, the testing of forensic and population genetics research.

关 键 词：线粒体DNA 单核苷酸多态 多重扩增产物片段长度多态性分析 

分 类 号：R394[医药卫生—医学遗传学]