Optimization of cloning and expression of 6His-tagged 196 aa length oncostatin M in E. coli

Abstract

A short, active form of oncostatin M protein consists of 196 aa and is 23 kDa protein. Using a full length open reading frame of this protein in pCMV type shuttle a PCR was performed and optimized in order to obtain the required form. A 6His tag was also added. As this active protein form is toxic to the most of the cells, accurate optimization of biosynthesis is required. A potent pT7a plasmid with double lacO sequence and early bacteriophage T7 promoter ensured highly stringent, controllable expression and its levels are further raised using medium optimization.