Cloning, Expression and Immunogenicity of Ferric Enterobactin Binding Protein Fep B from Escherichia Coli O157:H7

Overview

Journal Indian J Microbiol

Specialty Microbiology

Date 2012 Oct 27

PMID 23100780

Citations 1

Authors

Majid Alipour

Seyed Latif Mousavi Gargari

Iraj Rasooli

Affiliations

Soon will be listed here.

Abstract

The gene coding for ferric enterobactin binding protein from E. coli O157:H7 was amplifi ed. This gene was cloned and expressed as C-terminal His (6)-tagged protein. The SDS-PAGE analysis of the total protein revealed only two distinct bands, with molecular masses of 31kDa and 34kDa. The Ni-NTA chromatography purifi ed FepB and the osmotically shocked periplasmic fraction of IPTG induced cells showed only a single band of 31 kDa. Polyclonal mouse antibody was raised against the recombinant protein during 4 weeks after immunization. Western blot analysis of the recombinant FepB with mouse antiserum revealeda single band of 31 kDa. Identification and purification of FepB helped reveal its appropriate molecular mass. Polyclonal antibody raised against the recombinant protein reacted with bacterial FepB. The recombinant protein FepB could have a protective effect against E. coli O157:H7 and might be useful as an effective vaccine.

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