摘要
为研究丙肝病毒的复制机理,制备检测丙肝抗体的诊断试剂,需要大量纯化的各种丙肝病毒蛋白。我们在国内首次用DNA重组的办法,在大肠杆菌中表达了HCV部分NS4—NS5基因。在SDS—PAGE中,表达产物呈现一条约50kD的电泳带,表达蛋白约占菌体总蛋白的20％。该表达蛋白经分子杂交分析,可与抗-HCV非结构区抗体血浆发生特异反应。用部分提纯的表达蛋白制备的ELISA试剂,可特异地检测出我国抗—HCV诊断试剂参考品全部10个阳性样品,而对全部10个阴性样品均未发生特异反应,显示了该表达产物可用于制备抗—HCV诊断试剂的前景。
HCV antigen proteins are required in order to investiagte the replication of HCV and prepare the anti-HCV diagnostic reagents. We have expressed partial HCV NS4-NS5 gene in E. coli with DNA recombinant technique first time in China. The expressed products gave a single, 50 kD MW band in SDS-PAGE and showed positive reaction specially with anti-HCV non-structure antigen plasma in hybridization. The expression level was about 20% of tatol bacterial proteins. The Elisa reagent prepared with partial purified expression protein can recognize all 10 positive samples of national control panel of anti-HCV Elisa reagents in China and all 10 neagtive samples of that panel showed negative .results.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1995年第2期120-123,共4页
Chinese Journal of Microbiology and Immunology
