Cloning and expression of the gene of hepatitis C virus core in yeast

Objective: The function of HCV core is very complicated, the HCV core protein plays an important role in disease caused by HCV. In the report we investigate the gene expression of HCV core in yeast. Methods: PCR was performed to amplify the gene of HCV core from the plasmid pBRT M/HCV containing the whole fragment of HCV and the gene was cloned into pGEM-T vector. The gene of HCV core was cut from pGEM-T vector and cloned into yeast expression plasmid pGBKT7, then pGBKT7-core was transformed into yeast AH109. The yeast protein was isolated and analyzed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting assay. Results: HCV core gene was successfully cloned into pGBKT7. The results of SDS-PAGE and Western blotting assay showed (1) the molecular weight of the expressed product was about 22000Da.(2) HCV core protein was existed within yeast cells. Conclusion: The findings suggested that HCV core was successfully expressed in yeast system.