Research Journal of Biological Sciences
ISSN: Online 1993-6087ISSN: Print 1815-8846
Abstract
The gene coding for ferric enterobactin biding protein from E. coli O157:H7 was amplified. This gene was cloned and expressed as C-terminal His (6) -tagged protein. The SDS-PAGE analysis of the total protein revealed only 2 distinct bands, with molecular masses of 31 and 34 kDa. The Ni-NTA chromatography purified 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 revealed a single band of 31 kDa. Polyclonal antibody raised against the recombinant protein reacted with bacterial FepB. The successful production of antibody by periplasmic product of FepB gene can find a room for further research aimed at broad spectrum vaccines production. Identification and purification of FepB helped reveal its appropriate molecular mass. Reaction of the recombinant FepB antiserum with bacterial FepB finds its immunoactive contribution to protection against Gram negative bacteria harbouring the FepB protein. The serum of immunized mice was capable of protecting mice from live bacterial challenge. The recombinant protein FepB has a protective effect against E. coli O157:H7 on mice and might be useful as an effective vaccine.
How to cite this article:
Majid Alipour , Seyed Latif Mousavi Gargari , Iraj Rasooli and Shideh Montaser Kouhsari . Cloning, Expression and Immunoactivity of Periplasmic Binding Protein, FepB.
DOI: https://doi.org/10.36478/rjbsci.2008.1042.1047
URL: https://www.makhillpublications.co/view-article/1815-8846/rjbsci.2008.1042.1047