Role of HCV core protein in yeast twoª²hybrid system in reporter gene expression and activation

    LIU Min, ZHANG Wei, CHEN Tianª²Yan, LIN Shuª²Mei, LIU Jingª²Feng, YE Feng, ZHAO Yingª²Ren, ZHANG Shuª²Lin

    1Department of Infectious Diseases, First Affiliated Hospital, Medical College, Xi¬ðan Jiaotong University, Xi¬ðan 710061, China, 2Department of Microbiology, School of Basic Medicine, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To construct the bait vector of HCV core protein in yeast twoª²hybrid system and then to study its effect on the growth of yeast cells and the activation of reporter genes. METHODS: cDNA fragments encoding HCV core region were amplified by PCR and subsequently cloned into pUC19. After verified by sequencing, they were subcloned into the bait vector pGBKT7 of yeast twoª²hybrid system. These recombinant plasmids were transferred into AH109 and they activated the reporter genes. RESULTS: The fragments of HCV core protein were successfully obtained, which were not toxic to AH109 but could not activate the reporter genes. CONCLUSION: Yeast twoª²hybrid GAL4 system 3 can be utilized to fish HCV core regionª²interacting protein. HCV core protein does not have selfª²activating function. ......