HCV非结构基因NS4的基因工程表达

EXPRESSION OF HCV NONSTRUCTURE NS4 GENE IN E. COLI

用基因工程法在大肠杆菌中表达丙肝病毒(HCV)的部分非结构NS4基因,经分子杂交分析,表达产物可特异地与抗-HCV非结构区抗体反应。在SDS-PAGE中,表达产物呈现一条约49kD的融合蛋白带,约为菌体总蛋白的20％。用部分纯化的表达产物制备的ELISA试剂,可检测出国家丙肝诊断试剂参考品全部10个阳性样品中的8个,而对全部10个阴性样品均未出现假阳性反应,其检出率和P/C值均高于目前国内所用的5—1—1合成肽。显示了该表达产物用于丙肝诊断试剂及确证试剂的可能性。

A GST-HCV fused polypeptide encoding a segment of NS4 and of GST genes was expressed in E. coli under the control of ptac promotor first time in China. The expressed products gave a single band about 49kD MW in SDS-PAGE. This represented about 20% of total bacterial proteins. The hibrinization analysis showed that the expressed products could be recognized by the specific anti-HCV nonstructure antigen antibody in HCV patient's plasma, and the GST protein did not gave any nonspecific reaction with human plasma. The ELISA reagent prepared with partial purified expressed protein could recognize 8 of all 10 positive samples in the national control panel of anti-HCV ELISA reagents in China and all 10 negative samples of that panel showed negative .reaction. The result was better than that of chemical synthesized peptide 5-1-1.