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HomeAdvanced Materials ResearchAdvanced Materials Research Vol. 997Expression and Identification on the MPB83-MPB70...

Expression and Identification on the MPB83-MPB70 Fusion Gene of Mycobacterium bovis in Escherichia coli

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Abstract:

For raising the antigenicity of Mycobacterium bovis single antigen, fusion protein of two genes was acquired. The DNA fragments of mpb83 and mpb70 were fused by splicing by overlapping extension (SOE) polymerase chain reaction (PCR), and the fusion gene mpb83-mpb70 were cloned into pMD18-T vector, then we got the recombinant plasmid pMD-83-70. pMD-83-70 and pET28a (+) were digested by BamH I and EcoR I double enzymes. The purified pMD-83-70 fusion gene was subcloned into the expression vector pET28a (+), and the prokaryotic expression vector pET-83-70 was constructed. Plasmid containing pET-83-70 was transformed into competence Escherichia coli BL21 (DE3). The bacterium was induced by isopropyl-β-D-thiogalactoside (IPTG) and the lysates were loaded directly onto sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), approximately 41 kDa exogenous protein was observed on the SDS-PAGE. The protein was analyzed by using Western blotting, the results indicated that the protein was of antigenic activity of M. bovis. These results could serve as a basis for further studies on the usefulness of the fusion gene and its expression product in the development of subunit vaccine and DNA vaccine against bovine tuberculosis.

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Frontiers of Chemical Engineering, Metallurgical Engineering and Materials III

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Periodical:

Advanced Materials Research (Volume 997)

Pages:

210-214

DOI:

https://doi.org/10.4028/www.scientific.net/AMR.997.210

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Online since:

August 2014

Authors:

Yun Hang Gao*, Yan Liu, Ying Nan Zhang, Yan Yu, Chun Lei Liu, Hong Xia Ma, Chun Fang Wang, Bao Feng Luo, Xin Liu, Bing Ai, Xiu Yun Jiang

Keywords:

mpb83-mpb70 Fusion Gene, Mycobacterium bovis, Prokaryotic Expression

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© 2014 Trans Tech Publications Ltd. All Rights Reserved

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