In silico analysis of a chimeric fusion protein as a new vaccine candidate against Clostridium perfringens type A and Cl
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ORIGINAL ARTICLE
In silico analysis of a chimeric fusion protein as a new vaccine candidate against Clostridium perfringens type A and Clostridium septicum alpha toxins Ali Haghroosta 1,2 & Hossein Goudarzi 1 Reza Pilehchian Langroudi 2
&
Ebrahim Faghihloo 1 & Zohreh Ghalavand 1 & Mohammad Mahdi Ranjbar 3 &
Received: 7 September 2019 / Accepted: 8 June 2020 # The Author(s) 2020
Abstract In silico analysis is the most important approach to understand protein structure and functions, and the most important problem for designing and producing a fusion construct is producing large amounts of functional protein. Clostridium perfringens type A and Clostridium septicum produce alpha (plc) and alpha toxins respectively. C. perfringens can cause gas gangrene and gastrointestinal diseases. C. septicum can cause traumatic and non-traumatic gas gangrene. The aim of current research was in silico analysis of a chimeric fusion protein against C. perfringens type A and C. septicum alpha toxins. Firstly, the chimeric fusion gene was designed according to nucleotide sequences of C. perfringens type A alpha (KY584046.1) and C. septicum alpha (JN793989.2) toxin genes and then its fusion protein is constructed by amino acid sequences of C. perfringens type A and C. septicum alpha toxins. Secondly, online software was used to determine prediction of secondary and tertiary structures and physicochemical characteristics of the fusion protein. Finally, the validation of the fusion protein was confirmed by Rampage and proSA program. The designed fusion protein has 777 amino acids in length. TASSER server and physicochemical parameters are showed: C-score = − 2.68 and molecular weight = 87.9 KD respectively. Rampage and proSA software revealed the fusion protein is valid. Deposited accession number for the sequence of the fusion gene in the GenBank is MK908396. The designed fusion protein is valid and functional. Thus, the fusion gene could be used for clone and expression in a proper prokaryotic cell and also as a recombinant vaccine candidate. Keywords Clostridium perfringens . Clostridium septicum . Protein fusion . Vaccine . In silico
Introduction Clostridium perfringens is a Gram-positive, anaerobic, sporeforming bacteria. It causes anaerobic cellulitis, myonecrosis (gas gangrene), enteritis necroticans, and food poisoning in * Hossein Goudarzi [email protected] 1
Department of Microbiology, School of Medical, Shahid Beheshti University of Medical Sciences, Koodak-yar St, Daneshjoo Blvd, Velenjak, Chamran HWY, Tehran, Iran
2
Department of Anaerobic Bacterial Vaccines Production, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
3
Department of Poultry Virology Research, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
humans and gastrointestinal and enterotoxemic diseases in other animals (Dwivedi et al. 2015). C. perfringens based on the production of 4 major lethal toxins suc
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