CLONING AND EXPRESSION OF CLOSTRIDIUM BOTULINUM TYPE B BINDING DOMAIN E. COLI

Botulism is caused by botulinum toxin. The best way to avoid the neurotoxin
syndrome caused by BONT/B is to use recombinant vaccine made of BONT/B binding domain
because binding domain has sufficient epitops to stimulate immune response.
Materials & Methods: Initially BONT/B binding domain gene sequence were obtained from
GenBank. Then the primers were designed. PCR reaction was performed. Then gene was ligated to
pGEM-Teasy vactor. After verifying the transformation E.coli DH5α, subcloning was done. Pet28(a)+
vector was introduced to SDS-PAGE and Western blot confirmed production of the protein.
Results: The results obtained from PCR sequencing via IPTG induction and expression analysis by
SDS-PAGE and its confirmation by Western blot indicated the cloning and expression process
accuracy.
Conclusion: Finally, this method may be suitable for production of recombinant vaccines without any
side effects. Cloning and expression of this vaccine candidate were conducted successfully and its
immunization potential should be investigated.