Reports of Biochemistry and Molecular Biology
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2322-3480
2322-3480
10.61186/rbmb
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jalali
1398
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1
gregorian
2019
10
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Analysis of Methods to Improve the Solubility of Recombinant Bovine Sex Determining Region Y Protein
زیست شناسی ملکولی
Molecular Biology
مقالات اصلی
Original Article
<div style="text-align: justify;"><strong><em>Background:</em></strong> Inclusion body formation in <em>E. coli</em> 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) <em>E. coli</em> cells.<br>
<strong><em>Methods:</em></strong> 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.<br>
<strong><em>Results:</em></strong> 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.<br>
<strong><em>Conclusions:</em></strong> We found that modifications in temperature and IPTG and stabilizer concentrations affected rbSRY solubility.<br>
</div>
Cobsry, Inclusion Bodies, Recombinant Bovine SRY Protein, Solubility, Wtbsry.
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235
http://rbmb.net/browse.php?a_code=A-10-293-1&slc_lang=en&sid=1
Bijan
Soleymani
bijan.soleymani@kums.ac.ir
10031947532846006684
10031947532846006684
No
Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Ali
Mostafaie
amostafaie@kums.ac.ir
10031947532846006685
10031947532846006685
Yes
Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.