【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。更多还原

【Abstract】 [Objective] To investigate the polymorphisms in the promoter region of the tumour necrosis factor α(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 method.[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 Taqman MGB probes is simple,rapid,accurate,and automated,and will provide a useful tool for large-scale SNP association studies.更多还原