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Gene Synthesis> | … The routine chemicals were analytical grade (Sigma). 2.2. Construction of multi-copy expression vector. The vs30 sequence was optimized in accordance with the preferential codon usage for P. pastoris using the OptimumGene™ software and synthesized by Genscript™ … | Get A Quote |
Bacterial Expression System> | Get A Quote |
Vasostatin 30 (Vs30) is an active fragment derived from the N-terminal region (135-164aa) of human calreticulin (hCALR) and has the ability to inhibit angiogenesis. In this work, the expression of Vs30 was performed using a protease-deficient strain of the methylotrophic yeast Pichia pastoris. The vs30 gene was optimized for P. pastoris preferential codon usage and inserted into constitutive expression vector pGAPZαA. In addition, a multi-copy strain was obtained using an in vitro multimerization approach. The flask fermentation conditions were: culture volume of 25 mL in 250 mL baffled flasks at 28 °C, pH 6 and harvest time of 48 h. Up to 21.07 mg/L Vs30 were attained and purified by ultrafiltration with a 3... More