Comparison study of MeICT expression in pET۳۲-Rh and pET۳۲a cloning vectors

Backgrounds: Production of recombinant toxins with therapeutic applications is a challengingwork because of high number of disulfide bonds. Numerous expression systems are currentlyused to produce recombinant proteins. The pET system is one of the strongest expressionsystems in E. coli that includes different vectors. We previously designed pET۳۲Rh vector topurify recombinant proteins easier.Materials and Methods: In the present study, the expression of MeICT encoding fragment,which was isolated from Iranian yellow scorpion, was cloned in standard pET۳۲ and modifiedpET۳۲a-Rh vectors. The vectors were transformed and expressed in E. coli (BL۲۱). Differentexpression conditions such as temperature and time were investigated in this experiment. Then,the expression of MeICT was compared in the soluble and insoluble phases by SDS-PAGE.Finally, with the help of Image J software, the obtained results were quantified and statisticallycompared.Results: Our results showed that expression of gene in pET۳۲-Rh vector was comparable withstandard vector. The expression of MeICT was detected in high amount in soluble phase ofextracted protein for both vectors.Conclusion: Due to easier purification in pEt۳۲Rh compare to pET۳۲, the high expression ofMeICT showed that this vector could be a powerful system for the expression of toxin peptideswith multiple disulfide bonds.

کلیدواژه ها:

MeICT ، Recombinant toxins ، pET۳۲-Rh ، pET۳۲