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目的 : 探讨中国人iNOS基因是否存在单核苷酸多态性及其在中国人中的频率。方法 :DHPLC方法结合直接测序和PCR RFLP技术 ,在有代表性的中国人中进行筛查。结果 :在iNOS基因全部被测外显子中 ,有 6个外显子发现可疑双峰图型 ,可能存在杂合突变 ,对其测序后在iNOS基因第 16外显子发现C→T突变 (称为Tsp多态性 ) ,使编码的氨基酸由丝氨酸 (serine,Ser)改变为亮氨酸 (leucine ,Leu) ,设计PCR RFLP方法并监测全部测序样本 ,PCR RFLP与测序结果一致。在有代表性的 43例中国人中 ,Ser和Leu等位基因频率分别为 75 .5 8% (6 5 / 86 )和 2 4.42 % (2 1/ 86 ) ;三种基因型观察频率分别为Ser/Ser :6 2 .79% ;Ser/Leu :2 5 .5 8% ;Leu/Leu :11.6 3% ,与期望频率 (分别为 5 7.12 % ,36 .91%和 5 .96 % )差异无显著性 (P >0 .0 5 ) ,说明符合Hardy Weinberg遗传平衡定律。 结论 :中国人群中存在iNOS基因Tsp多态性 ,且频率较高
Objective: To investigate the existence of single nucleotide polymorphisms (SNPs) in Chinese iNOS gene and its frequency in Chinese. Methods: The DHPLC method combined with direct sequencing and PCR RFLP was used to screen out representative Chinese. Results: There were 6 exons found suspicious bimodal pattern in all tested exons of iNOS gene, there may be heterozygous mutation, after sequencing it found C → T mutation in exon 16 of iNOS gene Called Tsp polymorphism), the encoded amino acid was changed from serine (Ser) to leucine (Leu), the PCR RFLP method was designed and all the sequencing samples were monitored, and the PCR RFLP was consistent with the sequencing result. The allele frequencies of Ser and Leu were 43.8% (65/86) and 24.42% (21/86) respectively in 43 Chinese patients. The frequencies of the three genotypes were Ser / Ser: 62.79%; Ser / Leu: 25.55 8%; Leu / Leu: 11.6 3%, different from the expected frequencies (5 7.12%, 36.91% and 5.96%, respectively) No significant (P> 0.05), indicating that in line with the Hardy Weinberg law of genetic equilibrium. Conclusion: There is a polymorphism of iNOS gene Tsp in Chinese population, with a high frequency