解旋酶(helicase)在HCV基因组复制中负责RNA的解链,在复制中起着关键的作用。利用反转录PCR,从HCV患者血液中扩增出解旋酶基因,克隆入原核表达载体pProEXHTb,成功构建了HCV解旋酶原核表达质粒pProEXHTb-helicase。重组质粒转化DH5α大肠杆菌,经IPTG诱导表达后,表达产物经SDS-PAGE和Westernblot证实系解旋酶,并用镍离子亲和层析方法获得纯化解旋酶。 Helicase is responsible for the melting of RNA in HCV genome replication and plays a key role in replication. Using reverse transcription PCR, the helicase gene was amplified from the blood of HCV patients and cloned into prokaryotic expression vector pProEXHTb. The prokaryotic expression plasmid pProEXHTb-helicase of HCV helicase was successfully constructed. The recombinant plasmids were transformed into E. coli DH5α and induced by IPTG. The expressed products were confirmed by SDS-PAGE and Westernblot, and the helicase was confirmed by nickel ion affinity chromatography.