Cloning and expression of type D vaccine strain epsilon toxin gene in as a recombinant vaccine candidate.

Background And Objectives: a Gram-positive obligate anaerobic bacterium, is able to form resistant spores which are widely distributed in the environment. is subdivided into five types A to E based on its four major alpha, beta, epsilon and iota toxins. The aim of the present study was cloning and expression of type D vaccine strain epsilon toxin gene.

Materials And Methods: Genomic DNA was extracted and the epsilon toxin gene was amplified using DNA polymerase. The PCR product was cloned into pJET1.2/blunt cloning vector. The recombinant vector (pJETε) was sequenced using universal primers. At the next step epsilon toxin gene was subcloned into pET22b(+) expression vector and transformed into Rosetta (DE3) host strain.

Results: The recombinant protein has been expressed in Rosetta (DE3) cells after subcloning of gene (1008 bp) into the expression vector.

Conclusion: We concluded that Rosetta strain was suitable for the expression of recombinant epsilon toxin protein from pET22ε expression vector. This recombinant cell can be used for further research on recombinant vaccine development.