2004•Zhongguo yufang shouyi xuebaoRequires access
Expression of apoptin fusion gene in E.coli and preparation of its antibody
Shize Li
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Abstract
Recombinant expression plasmid pET-28 a-EGF-PⅡ-Apoptin was constructed according to recombinant plasmid pMD18-T-EGF-PⅡ-Apoptin. Then the recombinant was transformed into the host strain BL21(DE3) induced by IPTG when OD_(600) of the culture was about 0.6.The specific protein expressed (about 33KD) was detected by SDS-PAGE.The fusion protein was expressed at high level,amounting to 40 % of the total bacterial protein analyzed by thin-layer scan.After purified by electroelution in dialysis bags,the fusion protein was used to induce the production of polyclonal antibody in rabbits and ELISA detection showed the antigenicity of the fusion protein was satisfactory.Western blot showed the antiserum raised against the recombinant Apoptin in rabbits could react to the protein expressed specifically.