Bioinformatics study and evaluation of neurotoxin botulinum A optimized gene expression as a vaccine candidate

Backgrounds: Botulism is a deadly disease caused by neurotoxin, one of the seven types ofClostridium botulinum. Carboxyl part of the heavy chain neurotoxin Clostridium botulinum typeA (BoNT/A-Hc) has immunogenic properties that can be used as a recombinant candidatevaccine against botulism. The aim of this study was to investigate the bioinformaticsinvestigation and production of the recombinant neurotoxin-binding protein of BoNT /A.Materials and Methods: Nucleotide and protein sequences according to botA gene wereextracted from GeneBank and Uniprot databases. To increase the probability of proteinexpression, gene codons and various parameters affecting expression were optimized andthermodynamic analysis of mRNA structure was performed to evaluate the stability. Theantigenicity of the binding domain and its physicochemical properties were investigated. Thethird protein structure was predicted, the quality of the predicted structures was evaluated, linearand spatial epitopes were determined.Results: The Codon adaptation index (CAI) belonged to the natural gene WAS ۰.۲۸, while theoptimized gene had the index ۰.۹۶. The percentage of highly prevalent codons in the gene wasimproved to ۶۴. Thermodynamic analysis of the mRNA structure showed that the predictedstructure is stable. The third structure predicted based on the RaptorX server showed goodquality. Conformational and linear epitopes were seen in the protein.Conclusion: The results showed that the protein obtained could be a suitable immunogenicagainst Clostridium betulonium serotype A.

Keywords:

Neurotoxin botulinum A , Bioinformatics design , botA , Vaccine candidate