丙型肝炎病毒解旋酶在大肠杆菌中的表达研究

目的：在大肠肝菌中表达完整的丙型肝炎病毒的解旋酶。方法：用异硫氰酸胍法从HCV阳性的病人血清中提取RNA，经RT－PCR法扩增出NS3区的基因片段，将其克隆入PQE30表达载体中转化大肠杆菌XLI－Blue进行表达。表达出的蛋白用western blot方法鉴定。结果：扩增出的基因片段大小为1350bp，表达出的蛋白分子量为52KD，经Western blot鉴定为阳性。结论：成功地在大肠杆菌中表达出完整的HCV解旋酶蛋白，为HCV的诊断和疫苗研究奠定了基础。

Study on the expression of helicase of HCV in E.coli

Aim To expression the entire helicase of HCV in E.coli.Methods Aimed gene was extracted from the blood of HCV patient,and amplified by RT-PCR,the gene of helicase was coloned into the expressing vector PQE-30, then expressed in E.coil.M15.The expressed protein was indentified by western blot.Results The gene amplified by RT-PCR was identified about 1350 bp by the agrose gel electrophresis. The SDS-PAGE analiysis showed that the expressed protein is 52 KD.It was also identified by Western blot.Conclusions The entire helicase protein of HCV was expressed, which can be used to improve the detection rate and to the vaccine study of Hepatitis Cviurs.