Cloning and Expression of Glutaminase Gene glsA2 in Prokaryotic System

doi:10.7506/spkx1002-6630-201309029

Abstract

Abstract:

To test the activity of glutaminase from Bacillus subtilis BJ3-2, the gene glsA2 was cloned into prokaryotic
expression vector pET-32. The recombinant plasmid pET32a-glsA2 was then transformed into E.coli BL21 competent cells.
Optimum IPTG concentration and induction time for gene glsA2 expression were optimized. The result showed that the
expression was optimized with proper inducing conditions of 0.1 mmol/L IPTG for 6 h. Under these conditions, glutaminase
activity in supernatant of cell lysate reached 608.2 U/μg protein, which was 15 times higher than that of control.

Key words: glutaminase；glsA；prokaryotic expression；isopropyl &beta, -D-thiogalactopyranoside (IPTG) induction

CLC Number:

Q939.97

LU Biao，WU Yong-jun*，WU Yu-jun，LUO Xi，LIU Yan-min. Cloning and Expression of Glutaminase Gene glsA2 in Prokaryotic System[J]. FOOD SCIENCE, 2013, 34(9): 140-142.

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