IN SILICO FUSION OF ALPHA AND ALPHA TOXIN GENES OF CLOSTRIDIUM PERFRINGENS TYPE A AND CLOSTRIDIUM SEPTICUM

Background and Aim:Recombinant DNA technology is an in vitro molecular techniques to isolate and manipulate DNA fragments. Using this technique, construction chimeric molecules, called recombinant DNA molecules. The chimeric fusion protein technology represents the strategy to achieve rapid, cheap and efficient expression of proteins. Designing and producing a fusion construction is the most important problem of producing large quantities of functional protein. This construction should have all necessary components of a real gene. The aim of this study was designing and producing fusion of Clostridium perfringens types A and Clostridium septicum alpha-alpha toxin genes in silico.Methods:Clostridium perfringens type A alpha gene nucleotide sequence (cpa) and Clostridium septicum alpha gene (csa) were retrieved from GenBank. Then, to produce chimeric fusion protein, alpha-alpha fusion gene was designed. Secondary and tertiary structures and characteristics of the fusion protein was determined by online software. At last the fusion protein was validated by Rampage and ProSSA software.Results:Designed alpha-alpha fusion gene construction have 2334bp in length which nucleotides 1 to 1194 is alpha complete toxin of C.perfringens type A, nucleotides 1195 to 1230 (36 bp) is linker sequence which is optimized for E. coli and residue sequence is alpha activated toxin of C. septicum. Rampage and ProSSA software showed the fusion protein is validation and functional.Conclusion:The designed fusion gene construction is suitable to clone and expression in a suitable host cell, which could be used for producing of a recombinant alpha-alpha fusion protein vaccine.

کلیدواژه ها:

Clostridium perfringens ، Clostridium septicum ، Alpha toxin ، Fusion gene