Repository of Research and Investigative Information

Repository of Research and Investigative Information

Baqiyatallah University of Medical Sciences

Design and expression of a chimeric vaccine candidate for avian necrotic enteritis

(2017) Design and expression of a chimeric vaccine candidate for avian necrotic enteritis. Protein Engineering Design & Selection. pp. 39-45. ISSN 1741-0126

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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.

Item Type: Article
Keywords: Alpha toxin chimeric protein necrotic enteritis NetB TpeL clostridium-perfringens tpel broiler-chickens alpha-toxin netb toxin c-domain immunization performance protection challenge proteins Biochemistry & Molecular Biology Biotechnology & Applied Microbiology
Divisions:
Page Range: pp. 39-45
Journal or Publication Title: Protein Engineering Design & Selection
Journal Index: ISI
Volume: 30
Number: 1
Identification Number: https://doi.org/10.1093/protein/gzw060
ISSN: 1741-0126
Depositing User: مهندس مهدی شریفی
URI: http://eprints.bmsu.ac.ir/id/eprint/4728

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