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; Hossein Goudarzi; Ebrahim Faghihloo; Zohreh Ghalavand; Mohammad Mahdi Ranjbar; Reza Pilehchian Langroudi

doi:10.1007/s00580-020-03136-6

In silico analysis of a chimeric fusion protein as a new vaccine candidate against Clostridium perfringens type A and Clostridium septicum alpha toxins

Comp Clin Path. 2020;29(5):981-989. doi: 10.1007/s00580-020-03136-6. Epub 2020 Jul 14.

Authors

Ali Haghroosta^{1

2}, Hossein Goudarzi¹, Ebrahim Faghihloo¹, Zohreh Ghalavand¹, Mohammad Mahdi Ranjbar³, Reza Pilehchian Langroudi²

Affiliations

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.

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; In silico; Protein fusion; Vaccine.