巴基斯坦1a型HCV分离株核心基因哺乳动物表达载体的构建

B. Khubaib, M. Idrees, Abrar Hussain
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引用次数: 0

摘要

背景:细胞系是鉴定HCV病毒感染和繁殖的重要工具,通过建立表达核心蛋白的细胞系,我们可以探索HCV核心蛋白在HCC发生发展中的作用。结果:核心基因产物573bp PCR扩增及测序结果证实巴基斯坦分离株HCV为1A基因型。用限制性内切酶(Hindi III和EcoR I)对重组质粒进行酶切,分别得到5.5 kb和0.6 kb的片段,证明质粒具有核心基因。用核心基因重组哺乳动物载体瞬时转染Huh 7细胞系,提取核心蛋白21KDa,用抗核心单克隆抗体进行检测。结论:成功构建了在Huh 7细胞系中编码21KDa核心蛋白的巴基斯坦1A基因型核心基因的哺乳动物表达载体。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Construction of Mammalian Expression Vector of Core Gene of HCV of Pakistani Isolate Genotype 1a
Background: Cell lines are a valuable tool to identify the HCV virus infection and propagation and by establishing cell lines expressing core protein we can explore the role of HCV core protein in development of HCC. Results: The results indicate PCR amplification of 573bp product of core gene and sequencing confirmed the 1A genotype of HCV of Pakistani isolate. Then recombinant plasmid was digested with restriction enzymes (Hindi III and EcoR I) which gave fragments of 5.5 kb and 0.6 kb and It was a prove that our plasmid having a core gene. 21KDa core protein extracted from the transiently transfected Huh 7 cell line with recombinant mammalian vector of core gene was detected by using anti-core monoclonal antibody. Conclusion: we successfully constructed a mammalian expression vector of core gene of Pakistani isolate of genotype 1A that was encoding 21KDa of core protein in Huh 7 cell line.
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