title: Design and characterization of a chimeric multiepitope construct containing CfaB, heat-stable toxoid, CssA, CssB, and heat-labile toxin subunit B of enterotoxigenic Escherichia coli: a bioinformatic approach.
abstract: Enterotoxigenic Escherichia coli (ETEC) strains are the most common cause of bacterial diarrhea in children in developing countries and travelers to these areas. Enterotoxins and colonization factors (CFs) are two key virulence factors in ETEC pathogenesis, and the heterogeneity of the CFs is the bottleneck in reaching an effective vaccine. In this study, a candidate subunit vaccine, which is composed of CfaB, CssA and CssB, structural subunits of colonization factor antigen I and CS6 CFs, labile toxin subunit B, and the binding subunit of heat-labile and heat-stable toxoid, was designed to provide broad-spectrum protection against ETEC. The different features of chimeric gene, its mRNA stability, and chimeric protein properties were analyzed by using bioinformatic tools. The optimized chimeric gene was chemically synthesized and expressed successfully in a prokaryotic host. The purified protein was used for assessment of bioinformatic data by experimental methods.
authors.affiliation: Department of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Shahrak-e-Pajoohesh, Tehran, Iran.
authors.forename: Narges
authors.initials: N
authors.lastname: Zeinalzadeh
authors.forename: Ali Hatef
authors.initials: AH
authors.lastname: Salmanian
authors.forename: Ghasem
authors.initials: G
authors.lastname: Ahangari
authors.forename: Mahdi
authors.initials: M
authors.lastname: Sadeghi
authors.forename: Jafar
authors.initials: J
authors.lastname: Amani
authors.forename: S Zahra
authors.initials: SZ
authors.lastname: Bathaie
authors.forename: Mahyat
authors.initials: M
authors.lastname: Jafari
mesh_terms: D000595:Amino Acid Sequence; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D019295:Computational Biology; D054307:Enterotoxigenic Escherichia coli / Q000235:genetics / Q000276:immunology; D004768:Enterotoxins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D022361:Escherichia coli Vaccines / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D029968:Escherichia coli Proteins; D022361:Escherichia coli Vaccines; D011993:Recombinant Fusion Proteins
keywords: bioinformatics analysis; chimeric protein; enterotoxigenic Escherichia coli; immunogenicity