title: The codon-optimization of cfaE gene and evaluating its high expression capacity and conserved immunogenicity in Escherichia coli.
abstract: Enterotoxigenic Escherichia coli (ETEC) is the most common cause of children diarrhea in the world. Adhesion of ETEC to small intestine is an important virulence trait. One of the most prevalent colonization factors (CFs) in human is CFA/I fimbriae and CfaE which is the required binding factor for adhesion of ETEC to intestinal mucosa. We optimized cfaE gene codons according to codon bias of E. coli to achieve a high level of recombinant protein expression. The optimized gene was expressed in E. coli and rCFaE protein was used for mice immunization. Blocking activity of the obtained antibody was examined by microplate agglutination inhibition test. SDS-PAGE analysis indicated that the optimized sequence of cfaE produces a suitable amount of rCFaE in comparison with native gene sequence. This optimized rCFaE protein could induces strong humoral response in mice and the antibody obtained against rCFaE inhibited the adhesion of ETEC to human group A erythrocytes. It is concluded that codon optimization is a useful approach for obtaining large quantities of recombinant rCFaE protein. With regard to the results of hemagglutination inhibition test, codon optimization and increased production of recombinant protein expressed in E. coli did not affect the immunogenicity potential of CFaE.
authors: Department of Biology, Faculty of Basic Sciences, Imam Hussein University (IHU), Tehran, Iran.
authors: Maysam
authors: M
authors: Mansouri
authors: Seyed Jafar
authors: SJ
authors: Mousavy
authors: Zahra
authors: Z
authors: Ehsaei
authors: Shahram
authors: S
authors: Nazarian
authors: Mohammad Reza
authors: MR
authors: Zali
authors: Seyed Mohammad
authors: SM
authors: Moazzeni
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion / Q000187:drug effects / Q000276:immunology; D015153:Blotting, Western; D003062:Codon / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D054307:Enterotoxigenic Escherichia coli / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004912:Erythrocytes / Q000276:immunology / Q000378:metabolism; D004927:Escherichia coli Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control; D029968:Escherichia coli Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005260:Female; D036781:Fimbriae Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D056724:Immunity, Humoral / Q000276:immunology; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000276:immunology* / Q000378:metabolism
chemical_list: C559939:CfaE protein, E coli; D003062:Codon; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D036781:Fimbriae Proteins