Abstract:The purpose of this study is to construct the prokaryotic expression plsmid of SOD gene from Mycobacterium paratuberculosis and to express this gene in E.coli in which. pGEM-T-SOD and pET-28a(+) were digested in which double enzymes BamHⅠ and EcoRⅠ. The prokaryotic expression vector pET-28a-SOD was constructed by using the purified SOD gene that was subcloned into the expression vector pET-28a(+). Plasmid containing pET-28a-SOD was transformed into competence E. coli BL21 (DE3). The bacterium was induced by IPTG and its lysates were loaded directly onto SDS-PAGE. An approximately 26.5kDa exogenous protein was observed on the SDS-PAGE. The protein was analyzed by using Western-blotting and it had antigenic reactivity of Mycobacterium paratuberculosis. These results could serve as a basis for further studies on the usefulness of the gene and its expression product in the development of subunit vaccine and diagnostic reagent as bovine paratuberculosis.
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