丙型肝炎病毒C+E1区基因在原核细胞中的克隆与表达

Cloning and expressing of HCV/C+E1 gene in prokaryotic cell

将丙型肝炎病毒Ｃ＋Ｅ１区基因插入到原核高效表达载体ｐＢＶ２２１质粒中，构建了质粒ｐＢＶ２２１ＨＣＶ／Ｃ＋Ｅ１作为表达载体，然后，将含有该质粒的宿主大肠杆菌进行升温诱导表达ＨＣＶ／Ｃ＋Ｅ１区基因，并对表达产物进行了生物活性的检测。结果表明，插入到表达载体ｐＢＶ２２１中的ＨＣＶ／Ｃ＋Ｅ１基因片段能够得到有效的表达，表达产物主要为非融合蛋白形式存在于细胞中。

The C and close downstreamed El gene (HCV/C+E1 gene) which has been cloned from Hepatitis C virus (HCV) was inserted in plasmid pBV221, a high level expression vector,for construction of expression plasmid pBV221 HCV/C+E1.Having been transformed into E.coli and identified,the plasmid pBV221 HCV/C+E1 in host DH5 α strain was induced by temperature regulation to transcript and translate the HCV C+E1 gene.The biological activity of the co-expressed HCV/C+E1 protein was detected by ELISA.Results indicated that HCV/C+E1 gene in pBV221 expression vector can be effectively transcripted and translated.The C protein linked E1 protein which compose HCV/C+E1 non-fusion,chimera protein in E.coli cells,can be folded and modified properly to form their normal confirmation.Thus the antigenic activity of HCV/C+E1 protein is also maintained well,including their antigenicity against monoclonal Ab of anti-HCV/C protein.