In:
Advanced Materials Research, Trans Tech Publications, Ltd., Vol. 997 ( 2014-8), p. 210-214
Abstract:
For raising the antigenicity of Mycobacterium bovis single antigen, fusion protein of two genes was acquired. The DNA fragments of mpb83 and mpb 70 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 Bam H I and Eco R 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.
Type of Medium:
Online Resource
ISSN:
1662-8985
DOI:
10.4028/www.scientific.net/AMR.997
DOI:
10.4028/www.scientific.net/AMR.997.210
Language:
Unknown
Publisher:
Trans Tech Publications, Ltd.
Publication Date:
2014
detail.hit.zdb_id:
2265002-7
Bookmarklink