丙型肝炎病毒1a、1b和2a型E1E2原核质粒的构建和序列分析

发布时间：2018-04-24 13:19

本文选题：丙型肝炎病毒 + 包膜糖蛋白　；参考：《中国老年学杂志》2015年01期

【摘要】：目的构建陕西地方株丙型肝炎病毒(HCV)1a、1b和2a型包膜糖蛋白E1E2的原核质粒并测序比对。方法采用逆转录-巢式聚合酶链反应(RT-nest PCR)扩增获得HCV 1a、1b和2a型E1E2片段,与p MD-18 simple vector连接,转化感受态细菌DH5α,小量提取质粒DNA并测序。测序结果与Gene bank中其他标准序列比对以获取同源性数据。同时重点分析E2蛋白高变区(HVR)1和2氨基酸序列。MEGA 4软件绘制种系进化树图。结果成功扩增约2 kb的目的片段并构建原核质粒,测序结果正确。1b亚型E1E2的核苷酸和氨基酸序列的同源性最差。HCV E2的HVR1和2的序列比对显示,同种基因亚型在HVR 1的变异性最大,而HVR 2的序列变异性不显著。不同基因亚型的毒株在HVR 1和2的变异性极大,而1b亚型在HVR 1和2的变异性比1a和2a型突出。结论陕西地方株HCV 1b型E1E2区比1a和2a型更易发生突变,1b型E2的HVR变异性更显著。
[Abstract]:Objective to construct the primary nuclear particles of hepatitis C virus (HCV) 1a, 1b and 2A envelope glycoprotein E1E2 in Shaanxi, and to obtain HCV 1a, 1b and 2A E1E2 fragments by reverse transcription nested polymerase chain reaction (RT-nest PCR). Sequence. Sequencing results were compared with other standard sequences in Gene bank to obtain homologous data. At the same time, the phylogenetic tree of E2 protein hypervariable region (HVR) 1 and 2 amino acid sequence.MEGA 4 was plotted. The results were successfully amplified about 2 kb target fragments and constructed the original nucleolus, and sequenced the nucleotide and amino acid sequence of the.1b subtype E1E2. The sequence alignment of HVR1 and 2 of the worst homologous.HCV E2 showed that the variation of the homologous gene subtype was the largest in HVR 1, while the sequence variability of HVR 2 was not significant. The variants of the subtypes of the different genotypes were highly variable in HVR 1 and 2, while the 1b subtypes were more prominent in HVR 1 and 2 than in 1a and 2A type. Conclusion the HCV 1b type region of Shaanxi local strain is more than that And type 2A is more prone to mutation, and HVR variability of type 1b E2 is more significant.

【作者单位】：陕西中医学院医学技术系免疫及检验教研室;陕西绥德名州镇卫生院;
【基金】：陕西科学技术厅自然科学研究项目资助(No.SJ08C232)
【分类号】：R512.63

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