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丙型肝炎病毒(HCV)是一正链 RNA 病毒,基因全长约9400bp,一个大的开放读码框架编码约3010个氨基酸。包膜蛋白基因长约3000bp,分为 E_1和 E_2区。由于包膜蛋白的多变性,尤其是 E_2区的多变性,推测 HCV 的免疫逃避可能是 E_2变化所致,同时,E_2/NS_1也是 HCV 疫苗研究的重点,为此,我们对E_2/NS_1部分基因进行了克隆和序列分析。血清标本采自于抗-HCV 抗体呈强阳性的河南献血员。引物设计以毕胜利克隆株为模板,将其与美国和日本株比较后,选取保守区域,利用 Oligo 5.0和 Goldkey 软件设计,并在内引物两端加 BamH Ⅰ和 Pst Ⅰ位点。HCV RNA 提取参照文献[3]操作。提取 RNA 标本于94℃
Hepatitis C virus (HCV) is a positive-stranded RNA virus with a total length of about 9400 bp and a large open reading frame encoding approximately 3010 amino acids. The envelope protein gene is about 3000 bp in length and is divided into E 1 and E 2 regions. Because of the variability of envelope proteins, especially the variability of E 2 region, it is speculated that the immune evasion of HCV may be caused by the change of E 2. At the same time, E 2 / NS 1 is also the focus of HCV vaccine study. Therefore, Clones and sequence analysis were performed. Serum specimens were collected from Henan blood donors who were strongly positive for anti-HCV antibodies. Primer design was used as a template to clone the Bi Shengli cloning strain. After comparing with the American and Japanese strains, the conserved regions were selected and designed with Oligo 5.0 and Goldkey software. BamH I and Pst I sites were added to both ends of the primers. HCV RNA extraction reference [3] operation. RNA samples were extracted at 94 ° C