TY  - JOUR
T1  - Cloning, Expression and Immunoactivity of Periplasmic Binding Protein, FepB
AU - , Majid Alipour AU - , Seyed Latif Mousavi Gargari AU - , Iraj Rasooli AU - , Shideh Montaser Kouhsari 
JO  - Research Journal of Biological Sciences
VL  - 3
IS  - 9
SP  - 1042
EP  - 1047
PY  - 2008
DA  - 2001/08/19
SN  - 1815-8846
DO  - rjbsci.2008.1042.1047
UR  - https://makhillpublications.co/view-article.php?doi=rjbsci.2008.1042.1047
KW  - E. coli
KW  -FepB
KW  -ferric enterobactin
KW  -periplasmic binding protein
AB  - The gene coding for ferric enterobactin biding protein from <I>E. coli</I> 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 <I>E. coli</I> O157:H7 on mice and might be useful as an effective vaccine.
ER  -