2006•Zhongguo redai yixueRequires access
Expression of 25kDa outer membrane protein of Brucell in E.coli
Ming Li
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Abstract
Objective To clone and construct the recombinant expression plasmid for 25kDa Outer membrane protein(OMP25)of Brucella in E.coli.Methods The OMP25 gene of Brucella was amplified from genomic DNA and cloned into PMD-18 vector.It was then cloned into prokaryotic espression vector pGEX-4T-1 and identified by enzyme digestion and DNA sequencing.The recombinant plasmid was then transformed into E.coli HB101 and TOP10.The expression of OMP25 gene was induced by ITPG and the recombinant protein was identified by SDS-PAGE analysis.Results The results of DNA sequencing showed that the sequence and cloning site of the inserted OMP25 gene were correct,the recombinant expression plasmid pGEX-4T-1-Omp25 was constructed.Conclusion The OMP25 gene of Brucella are cloned and the recombinant OMP25 protein are successfully expressed in E.coli HB101.