作　　者： ; ; ; ; ;

机构地区： 深圳市疾病预防控制中心

出　　处： 《中山大学学报（医学科学版）》 2006年第1期51-54,62,共5页

摘　　要： 目的了解中国人群肿瘤坏死因子α(thetumournecrosisfactorα,TNF-α)基因启动子区多态性。方法随机选取20名深圳地区汉族健康体检者,采用两对引物PCR扩增TNF-α基因启动子区(-1389nt～+125nt),对PCR产物进行序列分析,寻找单核苷酸多态性位点(singlenucleotidepolymorphisms,SNPs)。采用TaqmanMGB探针建立了-857nt(C/T)位点的实时定量PCR(thereal-timePCR)分型方法,并对中国汉族、壮族、布依族,水族及苗族群体共1108份样本进行了基因分型。结果在启动子区(-1322nt～+67nt),发现6个SNP位点,即-885(A/G)、-863(C/A)、-646(G/A)、-648(G/A)、-568(G/C)和-857(C/T),其中位点-646nt(G→A)为新发现SNP。-885、-648及-568nt位点碱基虽然与Genbank不同,但测序的20个个体基因分型相同。中国人群-857nt(C/T)位点基因型频率分别为0.79(CC),0.19(CT)和0.02(TT),中国汉族、壮族、布依族,水族及苗族群体间无显著性差异。中国人群-857T等位基因频率为0.116,与文献报道的韩国人群相同,但比日本人群低。结论中国人群TNF-α基因启动子区单核苷酸多态性可能较保守。采用TaqmanMGB探针实时定量PCR技术对SNPs进行基因分型简便、快速及准确,易于自动化,为大规模的疾病相关性研究提供了有效的工具。 [Objective] To investigate the polymorphisms in the promoter region of the tumour necrosis factorα（TNF-α）gene from Chinese polulation.[Methods] Twenty Han healthy individuals in Shenzhen were selected randomly. The promoter region of the TNF-α gene in these 20 individuals from position -1389 nt to ＋125 nt was amplified by polymerase chain reaction （PCR） with two primer pairs. The PCR products were sequenced to scan the single nucleotide polymorphisms （SNPs）. The real-time PCR genotyping at -857 nt（C/T） position was established using Taqman minor groove binding （MGB） probes. Total 1108 samples from five Chinese populations, the Han, Zhuang, Buyi, Shui and Miao tribe groups were genotyped at -857 position using this methed.[Results] Six SNPs, - 885（A/G）, -863（C/A）, -646（G/A）, -648（G/A）, -568（G/C）, and -857（C/T）, were identified in the promoter region （- 1322 nt～＋67 nt）. Among these the -646 nt （G→A） SNP was a new one. Although the bases at -885,-648, and - 568 positions were different from the bases in Genebank sequence, the twenty individuals sequenced had the same genotypes at these three sites. The genotype frequencies of the SNP at site -857 nt（C/T） in Chinese population were 0.79, 0.19, and 0.02 for CC, CT, and TT, and there no difference among the five Chinese populations, the Han, Zhuang, Buyi, Shui and Miao tribe groups. The frequency of -857T allele in Chinese population was 0.116, similar to that reported in Korean, but lower than that in Japanese. [ Conclusion ] The SNPs in the promoter region of TNF-α gene from Chinese population may be conserved. The real-time PCR technology for SNPs genotyping using TaqmanMGB probes is simple, rapid, accurate, and automated, and will provide a useful tool for large-scale SNP association studies.

关 键 词： 多态性 肿瘤坏死因子 单核苷酸多态性 实时定量 探针

领　　域： [生物学]