title: Design and expression of a chimeric vaccine candidate for avian necrotic enteritis.
abstract: Necrotic enteritis is an economically important disease of poultry mainly caused by Clostridium perfringens The bacteria release multiple toxins of which NetB, alpha toxin and TpeL have been reported to play important roles in pathogenicity and/or severity of the disease. In this study, the sequence of clostridial toxins NetB, alpha toxin and TpeL were analyzed using bioinformatics tools to determine protein domains with high immunogenicity factor. Several chimeric trivalent proteins consisting of the immunogenic regions of the three toxins were designed and evaluated. The separate regions were fused together using rigid linkers. Based on a modeled tertiary structure, a proper combination was selected and expressed in a bacterial host (Escherichia coli) and successfully purified. The expression of the chimeric protein was further verified by western blotting. The ability of the immunized serum in recognizing each individual subunit of the chimeric protein was also examined. Circular dichroism was used to evaluate the predicted secondary structure of the chimeric protein. In vitro potency test demonstrated that the serum from a rabbit immunized with the chimeric protein is able to partially neutralize Alpha toxin, hence the construct can potentially be used as a vaccine against C. perfringens.
authors.affiliation: Department of Basic Sciences, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran 1971653313, Iran.|Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran 1497716316, Iran.
authors.forename: Amin
authors.initials: A
authors.lastname: Rostami
authors.affiliation: Department of Physiology, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran 1971653313, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Goshadrou
authors.affiliation: Clostridia Specialized Research Laboratory, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj 3197619751, Iran.
authors.forename: Reza Pilehchian
authors.initials: RP
authors.lastname: Langroudi
authors.affiliation: Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran 14115-331, Iran.
authors.forename: S Zahra
authors.initials: SZ
authors.lastname: Bathaie
authors.affiliation: AffyCell Biologics Inc., Toronto L3T 1K6, Canada.
authors.forename: Ali
authors.initials: A
authors.lastname: Riazi
authors.affiliation: Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran 1435916471, Iran Jafar.amani@gmail.com.
authors.forename: Jafar
authors.initials: J
authors.lastname: Amani
authors.affiliation: Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran 1497716316, Iran Jafar.amani@gmail.com.
authors.forename: Gholamreza
authors.initials: G
authors.lastname: Ahmadian
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D001428:Bacterial Vaccines / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D002645:Chickens / Q000382:microbiology*; D003001:Cloning, Molecular; D003016:Clostridium perfringens / Q000276:immunology / Q000502:physiology*; D004751:Enteritis / Q000517:prevention & control*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007114:Immunization; D008958:Models, Molecular; D000072417:Protein Domains; D015202:Protein Engineering / Q000379:methods*; D017433:Protein Structure, Secondary; D011817:Rabbits
chemical_list: D001427:Bacterial Toxins; D001428:Bacterial Vaccines
keywords: Alpha toxin; NetB; TpeL; chimeric protein; necrotic enteritis