EXPRESSION OF GLUTATHIONE S-TRANSFERASE-CARDIOTOXIN FUSION PROTEIN INESCHERICHIA-COLI

We report here the construction of cardiotoxin V gene, from cobra snak e venom (Naja naja atra), by chemically synthesized oligonucleotides a nd its expression as a glutathione S-transferase-cardiotoxin fusion pr otein in the inclusion bodies of Escherichia coli. The expression of c ardiotoxin fusion protein with a yield of about 35 mg/liter culture wa s confirmed by highly specific anti-peptide antibodies generated again st the unique amino acid residues located at the tip of loop II of car diotoxin V. Since the fusion protein can be easily treated by CNBr to free the toxin moiety, as revealed by immunoblotting of the cleaved pr otein, the results provide an avenue for future structural and functio nal studies of cardiotoxin molecules.