HIGN LEVEL EXPRESSION OF HEPATITIS C VIRUS CORE GENE IN E. COLI
- Author:
Hua LI
;
Chengen PAN
;
Wuke CHEN
;
Xinlu WANG
;
Quanying WANG
;
Guangxiao YANG
- Publication Type:Journal Article
- Keywords:
hepatitis C virus;
core protein;
gene;
viral;
antigen
- From:
Journal of Pharmaceutical Analysis
2000;12(2):142-145
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo express the hepatitis C virus (HCV) core gene in E . coli on a high level. Methods The cDNA coding for HCV core protein was amplified by polymerase chain reaction (PCR). The PCR product was purified and digested with restriction enzymes and inserted into the downstream of PRPL promoter of a high-level ex- pression vector pBV220. HCV core gene was expressed in E. coli in a non-fused form. The expression protein was analysed by SDS-PAGE , and its immunoactivity was tested by ELISA. Results Sequence analysis of the amplified PCR products confirmed that we have successfully cloned and expresssed the intact core protein of HCV. SDS-PAGE showed that a specific protein with a molecular weight of 21kDa at a level of 14. 0％ of the total bacterial proteins ap- peared in bacteria harboring pBV/HCVCore, while this protein was absent in the control bacteria harboring pBV220. The results of enzyme immunoassay analysis showed that this protein could be specifically recognized by the HCV pos- itive sera from patients with hepatitis C . Conclusion The intact HCV core protein was successfully expressed in E . coli in a non-fused form on a high level, and its immunoactivity was high.
