TY - JOUR T1 - Analysis of Methods to Improve the Solubility of Recombinant Bovine Sex Determining Region Y Protein TT - JF - rbmb JO - rbmb VL - 8 IS - 3 UR - http://rbmb.net/article-1-344-en.html Y1 - 2019 SP - 227 EP - 235 KW - Cobsry KW - Inclusion Bodies KW - Recombinant Bovine SRY Protein KW - Solubility KW - Wtbsry. N2 - Background: Inclusion body formation in E. coli is a significant problem in recombinant protein production. The aim of this study was to improve the solubility of recombinant bovine sex determining region Y protein (SRY) in BL21 (DE3) E. coli cells. Methods: In this research two recombinant bovine SRY (rbSRY) sequences were analyzed; these were wild-type SRY (wtbSRY) and codon-optimized SRY (cobSRY). Their expression in various culture conditions was examined; these differences included IPTG concentrations, temperatures, and media stabilizers. Results: IPTG and temperature significantly affected rbSRY solubility (P < 0.001). The optimum IPTG concentration and temperatures for wtbSRY and cobSRY induction were 0.3 mM at 27 and 32 °C, respectively. In addition, arginine and sorbitol concentrations significantly affected rbSRY solubility (P < 0.01). Solubility of rbSRY protein was highest from the cobSRY construct in the presence 0.2 M arginine and 0.3 M sorbitol. The highest inclusion body production occurred with high glucose concentrations. Conclusions: We found that modifications in temperature and IPTG and stabilizer concentrations affected rbSRY solubility. M3 ER -