丙型肝炎病毒E1，E2/NS1，NS5蛋白基因在大肠杆菌内的表达及其产物的初步应用

目的在体外表达及纯化ＨＣＶ不同功能区的病毒蛋白，用于研究ＨＣＶ各功能区抗体的临床意义。方法将ＨＣＶＥ１，Ｅ２／ＮＳ１，ＮＳ５区的部分基因片段克隆到表达质粒ＰＭＡＬ－ＣＲＩ中ＭＢＰ编码基因的下游，在大肠杆菌中进行表达。结果表达的融合蛋白ＭＢＰ－Ｅ１，ＭＢＰ－Ｅ２／ＮＳ１，ＭＢＰ－ＮＳ５的分子量分别为５７０００、６５０００、６２０００，ＷｅｓｔｅｒｎＢｌｏｔ分析表明这些融合蛋白具有ＨＣＶ的抗原活性。将这三种融合蛋白纯化后作为抗原检测了丙型肝炎患者血清９５份，其中抗－ＨＣＶＥ１阳性率为１０．５％，抗－ＨＣＶＥ２／ＮＳ１为６３．２％，抗－ＨＣＶＮＳ５为５３．７％。结论本实验表达的ＨＣＶ融合蛋白在ＨＣＶ感染的血清学诊断中具有一定的价值。

Expression of E1,E2/NS1,NS5 genes of hepatitis C virus in E.coli and preliminary use of the expressed proteins

Objective In order to study the clinical significance of various antibodies against different regions of HCV genome, the proteins in different regions of HCV were expressed and purified in vitro. Methods Fragments of E1,E2/NS1 and NS5 of HCV genes were cloned into expression vector PMAL CRI and the fusion proteins MBP E1,MBP E2/NS1 and MBP NS5 were expressed in E.coli. Results The molecular weights of MBP E1,MBP E2/NS1 and MBP NS5 were 57000 ,65000 and 62000 respectively. Western blot analysis showed these fusion proteins presented the specific HCV antigenicity and were used to detect 95 serum specimens of hepatitis C patients by ELISA after purification. The positive ratio of anti HCV E1,anti HCV E1/NS1 and anti HCV NS5 were 10.5%,63.2% and 53.7% respectively. Conclusion These fusion proteins are useful in the serodiagnosis of HCV infection.