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目的探讨中国大陆1b型丙型肝炎病毒(HCV)3’非编码区(3’UTR)一级结构的变异规律,为进一步研究其在HCV复制、翻译中的调控机制、开发新的抗HCV药物奠定基础。方法利用逆转录套式聚合酶链反应(RT-PCR)限制性内切酶长度多态性分析(RFLP)初步筛选出1b型HCV感染者,采用半套式RT-PCR法扩增出约400 bp的cDNA片段,克隆测序。结果获得了全长1b型HCV 3’端序列,发现终止密码子存在两种突变,由TGA突变为TAA或TGG,从而丧失了终止密码子功能,可导致NS5B的翻译不能及时终止。结论中国大陆1b型丙型肝炎病毒终止密码子存在多种突变,其中TGG突变可导致NS5B区延长,3’UTR缩短;该发现对了解HCV的复制和翻译机制可能有一定的理论和实践意义。
Objective To explore the variation of the primary structure of 3 ’untranslated region (HCV) 3’ UTR in the Chinese mainland. In order to further investigate its regulatory mechanism in HCV replication and translation, a new anti-HCV drug was developed Lay the foundation. Methods The HCV genotype 1b was screened by reverse transcription-polymerase chain reaction (RT-PCR) restriction endonuclease length polymorphism (RFLP), and amplified by semi-set RT-PCR bp cDNA fragment, cloned and sequenced. Results The full-length HCV 3 ’end sequence was obtained and found that there were two mutations in the stop codon, from TGA to TAA or TGG, which resulted in the loss of stop codon function and the inability to terminate the NS5B translation in time. CONCLUSION: There are many mutations in the stop codon of hepatitis C virus type 1b in Chinese mainland. TGG mutation leads to prolongation of NS5B region and shortening of 3’UTR. This finding may have theoretical and practical significance for understanding the mechanism of HCV replication and translation.