Expressions of different deletion mutated HCV core protein genes in E.coli

    SHEN Caiª²Fei, BAI Xueª²Fan, MOU Danª²Lei£¬HUANG Changª²Xing

    PLA Center for Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi¬ðan 710038, China

    ¡¾Abstract¡¿ AIM: To construct the recombinant plasmids expressing fullª²length HCV core protein gene and 3 different deletion mutated hepatitis core protein genes and to express them in E.coli. METHODS: Entire HCV core protein gene and 3 different deletion mutated gene fragments from HCV core protein which respectively coded 1-191aa, 1-59aa plus 81-191aa, 1-169aa, 1-59aa plus 81-169aa were amplified by PCR and cloned into expression vector pRSETª²A, thus forming different recombinant plasmids (pRSETª²Aª²C573, pRSETª²Aª²C510, pRSETª²Aª²C507, pRSETª²Aª²C444), which were transformed into E.coli BLª²21 (DE3) pLysS and induced by IPTG. SDSª²PAGE and Western blot were used to test and identify expressed 6¡ÁHisª²fusion proteins. RESULTS: Restriction analysis and sequencing confirmed that the 4 recombinant plasmids expressing entire HCV core protein gene and 3 different deletion mutated hepatitis core protein genes were successfully constructed. Both SDSª²PAGE and Western blot analysis showed entire HCV core protein gene and 3 different deletion mutated hepatitis core protein genes were expressed, and the respective molecular weights of the products were about 21¡Á103, 19¡Á103, 19¡Á103 and 16.5¡Á103. The fusion protein production accounted approximately for 20% of total bacterial protein. CONCLUSION: Different deletion mutated HCV core protein genes were expressed efficiently in E.coli. ......