丙型肝炎病毒NS3区优势表位基因的克隆和高效表达研究

Molecular Cloning and high cxpression of HCV NS3 gene with an Unfused Protein in E. Coli cells

丙型肝炎病毒（ＨＣＶ）的Ｃ３３ｃ基因编码蛋白是ＨＣＶ抗体检测中所需的重要抗原。应用打点杂交法得到了来源于３例病人的８个Ｃ３３ｃ的克隆，并从中挑选出能在大肠杆菌中高效表达的克隆─—ｐＫＨ２６。该重组质粒中的插入片段与ＨＣＶ－Ｊ的核苷酸同源性为９２．６％，所编码氨基酸的同源性为９５．９％，属于中国主要的ＨＣＶ流行株，它可通过温度变化这种简单、方便、经济的方式进行诱导表达，得到以天然蛋白形式存在的具有良好免疫学活性的表达产物，表达蛋白产量可占整个细菌蛋白的１５％以上。由于不含有融合蛋白，用于抗－Ｃ３３ｃ的检测特异性高，是建立诊断ＨＣＶ感染方法的良好原材料。

epatitis C Virus HCV) is a major causative agent of post-transfusion as well as sporadic parentally transmitted NANBH throughout the world. C33c gene of HCV is an important gene for detec- tion of HCV antibody. 8 clones of HCV C33c gene from 3 patients was gotten, A recombinant plasmid pKH26 had been screened, and was expressed efficiently in pop21 36 E. coli cells. The inserted fragment included 810 nucleotides of HCV NS3 gene, which could encod 270 amino acids with methionine in the first position. Its nucleotide homology with HCV-Ⅱ subtype is 92.6%,aminoacid homology is 95.9%,The recombinant plasmid can expressed efficiently with unfused protein by temperature induction .The expressed C33c represented more than 15% of total bacterial proteins.