Heterologous expression, purification, and refolding of SRY protein: role of l -arginine as analyzed by simulation and p
- PDF / 1,077,037 Bytes
- 9 Pages / 595.276 x 790.866 pts Page_size
- 31 Downloads / 155 Views
ORIGINAL ARTICLE
Heterologous expression, purification, and refolding of SRY protein: role of l‑arginine as analyzed by simulation and practical study Bijan Soleymani1 · Ebrahim Barzegari1 · Kamran Mansouri1 · Keyvan Karami1 · Pantea Mohammadi1 · Sarah Kiani1 · Narges Moasefi1 · Mehdi Sharifi Tabar2 · Ali Mostafaie1 Received: 22 April 2020 / Accepted: 13 July 2020 © Springer Nature B.V. 2020
Abstract Escherichia coli is a widely-used cell factory for recombinant protein production, nevertheless, high amount of produced protein is seen in aggregated form. The purpose of this study was to improve the solubility of recombinant bovine sexdetermining region Y protein (rbSRY) by exploring the effect of temperature, inducer, and water-arginine mixed solvent. Codon-optimized rbSRY expressed in Rosetta-gami B (DE3) pLysS and purified by NI–NTA His-select affinity chromatography in the native and denaturing conditions. A three-dimensional model of SRY was built and studied through molecular dynamics simulations in water and in the presence of l-arginine as co-solvent. Results indicated the significant effects of temperature and IPTG concentration (P
Data Loading...
