Abstract：To improve antigenicity of the simple protein, Mycobacterium bovis MPB51 and MPB70 were fused using (Gly4Ser)3 to obtain a fusion protein MPB51-MPB70. The mpb51 and mpb70 genes were fused by splicing overlapping extension(SOE) polymerase chain reaction(PCR) to acquire the fusion gene mpb51-mpb70 which was then cloned into pMD19 vector to construct recombinant plasmid pMD-51-70. After enzyme digestion by BamH I and EcoR I, the purified fusion gene was subcloned into the expression vector pET28a(+). The expression of mpb51-mpb70 was analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and the antigenic activity was determined by Western-blotting. Indirect-ELISA was used to compare the antigenicity among MPB51, MPB70 and MPB51-MPB70. A 1359bp fusion gene was amplified and the recombinant plasmid pMD-51-70 was constructed successfully. The expressed fusion protein, approximately 45 ku, exhibited a reaction activity with bovine tuberculosis serum. Besides, the reaction activity was better than that of solo protein. These results indicated that the MPB51-MPB70 fusion protein was obtained successfully.
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2018, Vol. 34 
