庚型肝炎病毒基因在大肠杆菌中表达的初步研究

Preliminary Studies of Prokaryotic Expression of Hepatitis G Virus Gene

利用原核表达载体 pRSET或 (和 )pGEX在大肠杆菌内表达了覆盖庚型肝炎病毒 (HGV)C NS3和NS5区的多段基因。CE1、E2、NS3、NS5及NS3 NS5嵌合基因等的 8段基因均有高效表达 ,各重组蛋白产量与菌体总蛋白之比在 10 %～ 35 %之间。对以上重组蛋白进行免疫学筛选 ,证实其中 7个重组蛋白均具免疫学活性 ,在一定程度上确定了重组HGV抗原表位的分布 。

The prokaryotic expression vector pRSET of (and) pGEX was used to express overlapping clones emcompassing hepatitis G virus Core-NS3 and NS5 in E. coli. CE1、E2、NS3、NS5 and chimeric NS3-NS5 gene could produce recombinant proteins efficiently. The ratio of every recombinant protein to total bacteria proteins varied from 10% to 35%. All recombinant proteins were immuoscreened, the results showed that seven of them were immuoreactive. To some extent,we identified the epitopic distribution of recombinant HGV antigens and made a solid foundation for the research of HGV serology and immunological diagnostic reagents.