Expression of Gankyrinª²GST fusion protein in E.coli and preparation of its specific polyclonal antibody

    YANG Zhaoª²Xu1, DOU Keª²Feng1, LU Fan2, ZHAO Zhongª²Liang2

    1Department of Hepatobiliary Surgery, Xijing Hospital, 2Department of Biochemistry and Molecular Biology, School of Basic Medicine, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To express Gankyrin in E. coli and to prepare antibody against it. METHODS: The core fragment of Gankyrin was cloned into plasmid pGEXª²4Tª²2 containing glutathione sª²transferase (GST) fusion protein gene. Following the restriction enzyme digestion analysis and sequencing, pGEXª²4Tª²Gankyrin was transformed into E. coli DHª²5¦Á. GSTª²Gankyrin fusion protein was expressed under IPTG induction and the Gankyrin protein was used to immunize New Zealand rabbits. The specific antibody against Gankyrin was identified by ELISA and Western blot. RESULTS: GSTª²Gankyrin fusion protein was expressed in E.coli and its specific polyclonal antibody was obtained. CONCLUSION: The successful expression of GSTª²Gankyrin fusion protein in E.coli and the preparation of its specific polyclonal antibody will facilitate the study of the biological functions of Gankyrin. ......