In:
Advanced Materials Research, Trans Tech Publications, Ltd., Vol. 884-885 ( 2014-1), p. 498-502
Abstract:
Based on splicing by overlapping extension (SOE) polymerase chain reaction (PCR) ,the ag85a and mpb70 were amplified and the fusion gene ag85a-mpb70 were cloned into pMD18-T vector, and then we got the recombinant plasmid pMD-85a-70. pMD-85a-70 and pET28a (+) were digested by Bam HI and Eco RI double enzymes. The purified ag85a-mpb70 fusion gene was subcloned into the expression vector pET28a (+),and the prokaryotic expression vector pET-85a-70 was constructed. Plasmid containing pET-85a-70 was transformed into competence Escherichia coli BL21(DE3).The bacterium was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) and analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), approximately 49 kDa fusion protein was observed on the SDS-PAGE. The protein was analyzed by using Western-blotting. The results indicated that Ag85A-MPB70 was of antigenic activity of Mycobacterium 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 novel vaccine against bovine tuberculosis.
Type of Medium:
Online Resource
ISSN:
1662-8985
DOI:
10.4028/www.scientific.net/AMR.884-885
DOI:
10.4028/www.scientific.net/AMR.884-885.498
Language:
Unknown
Publisher:
Trans Tech Publications, Ltd.
Publication Date:
2014
detail.hit.zdb_id:
2265002-7
