Codon Optimization and Comparison of Recombinant Human β-NGF Expression in BL21(DE3)pLysS and BL21(DE3) Bacterial Hosts

Beta Nerve growth factor (β-NGF) was first known for its vital role in development and survival of neurons. This protein, which belongs to the neurotrophin family, plays a considerable role in the treatment of neurodegenerative diseases such as Alzheimer’s disease (AD). As a result of its characteristic, recombinant human β-NGF (rhβNGF) was expressed in two different strains of E.coli for the first time in Iran. β-NGF has three disulfide bonds in its native and functional structure, so the cytoplasmic reducing environment of E.coli is not appropriate for its expression. Therefore, the oxidative space of periplasm for production of correctly folded β-NGF was considered. In this study, hβNGF cDNA obtained from NCBI data bank after codon optimization and subcloning in pET39b() vector containing DsbA gene was transformed (using heat shock) to BL21(DE3)pLysS and BL21(DE3) strains of E.coli. Co-expression occurred via induction of promoter with 1 mM of IPTG. Consequently, extracted proteins from these two strains were compared with each other with SDS-PAGE and Dot blot techniques. The data shows βNGF expression especially in BL21(DE3) strain of E.coli.