应用等位基因特异性PCR检测胆固醇酯转运蛋白基因突变  

Detection of the CETP gene mutation by AS-PCR

作　　者：周代锋[1] 蔡望伟[1] 云美玲[2] 张勇[3] 习隽丽[3] 王镇[3] 

机构地区：[1]海南医学院生物化学教研室,海南海口571101 [2]海南医学院附属医院心内科,海南海口570102 [3]海南省老年病医院,海南海口571100

出　　处：《海南医学院学报》2009年第7期693-697,701,共6页Journal of Hainan Medical University

基　　金：海南省自然科学基金立项课题(NO.琼科函[2004]37号-1);海南医学院苗圃基金资助课题(NO.海医学[2005]47号-1)~~

摘　　要：目的:建立检测胆固醇酯转运蛋白(cholesterol ester transfer protein,CETP)基因6种常见突变的等位基因特异性PCR技术。方法:应用针对CETP基因TaqIB(G→A)、I405V(A→G)、D442G(A→G)、R451Q(G→A)、A373P(G→C)和I14A(G→A)这6种常见的突变位点设计的等位基因特异性PCR技术,对海南汉、黎族人群中CETP基因突变类型进行了检测,同时对经上述等位基因特异性PCR检测的样本进行序列测定。结果:在海南汉、黎族人群中,TaqIB(G→A)突变位点可检测出GG、GA、AA3种基因型,I405V(A→G)突变位点可检测出AA、AG、GG3种基因型,D442G(A→G)突变位点可检测出AA、AG2种基因型,但在海南汉、黎族人群中未检测到R451Q(G→A)、A373P(G→C)和I14A(G→A)3种突变类型,用等位基因特异性PCR鉴定的CETP基因突变的基因分型结果与序列测定结果完全符合。结论:等位基因特异性PCR技术操作简便,重复性和稳定性好,可作为鉴定CETP基因突变类型的可行方法。Objective： To establish the allele specific polymerase chain reaction （AS-PCR） technique for the detection of 6 normal mutations of cholesterol ester transfer protein （CETP） gene. Methods： Designed the AS-PCR system according to the 6 mutations of CETP gene, TaqlB （ G→A）, I405V （ A→G）, D442G （ A→G）, R451Q （G→A）, A373P （G→C） and I14A （G→A）. Detected the mutations in Li and Han nationalities of Hainan province by the designed method and sequenced partial mutation samples. Results： 3 genotypes were successfully detected in TaqIB （G→A） mutation site which were GG, GA, AA, 3 genotypes were successfully detected in I405V （A→G） mutation site which were AA, AG and GG, and 2 genotypes were successfully detected in D442G（ A→G） mutation site which were AA and AG, while only the wild genotype could be detected separately in R451 Q （G→A） ,A373P（G→C） and I14A（G→A） mutation sites which was GG. All the results above provided by the AS-PCR were exactly consistent with the sequencing results. Conclusion ： The AS-PCR with advantages of stability and simplify is an effective method for investigation of the mutations of CETP gene.

关 键 词：胆固醇酯转运蛋白基因 等位基因特异性 聚合酶链反应 基因分型 

分 类 号：Q549[生物学—生物化学] Q503