Volume 8, Issue 3 (Vol.8 No.3 Oct 2019)                   rbmb.net 2019, 8(3): 227-235 | Back to browse issues page

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Soleymani B, Mostafaie A. Analysis of Methods to Improve the Solubility of Recombinant Bovine Sex Determining Region Y Protein. rbmb.net. 2019; 8 (3) :227-235
URL: http://rbmb.net/article-1-344-en.html
Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Abstract:   (739 Views)
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.
 
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Type of Article: Original Article | Subject: Molecular Biology
Received: 2019/04/27 | Accepted: 2019/05/26 | Published: 2020/01/24

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