Prokaryotic expression and activity analysis of thioredoxin reductase from Bacillus subtilis

doi:10.3969/j.issn.1004-1524.2016.12.14

›› 2016, Vol. 28 ›› Issue (12): 2054-2059.DOI: 10.3969/j.issn.1004-1524.2016.12.14

• Plant Protection • Previous Articles Next Articles

Prokaryotic expression and activity analysis of thioredoxin reductase from Bacillus subtilis

CHEN Hong-na¹, LI Jian-wen¹, JIANG Ao¹, SUN Wen-xiu^1,2,*

1. College of Life Sciences, Yangtze University, Jingzhou 434025, China;
2. Institute of Root Biology, Yangtze University, Jingzhou 434025, China

Received:2016-04-05 Online:2016-12-15 Published:2017-01-05

Abstract

Abstract: NADPH dependent thioredoxin reductase is a flavoprotein containing selenium, which plays an important role in cell proliferation and differentiation and maintaining cell oxidation-reduction equilibrium. The TrxR gene from Bacillus subtilis was amplified and subcloned into pET-28a(+) expression vector. Subsequently, the plasimid was transformed into Escherichia coli BL21(DE3). And then expression and purification of TrxR were performed. Results showed that the TrxR gene from B. subtilis strain BS04 was 1 011 bp in length, which encoding 336 amino acids; the amino acid identity between the gene we cloned and the sequence deposited in GenBank was about 99%. SDS-PAGE and Western-blotting analysis suggested that the molecular weight of the fusion protein was about 37 ku which was in accord with the estimated. Moreover, the kinetic parameters K_m and K_cat of the recombinant protein were determined to be 3.06 mmol·L^-1 and 324.71 min^-1 using DTNB assay. All above results provide good opportunity for our further study on the gene function.

Key words: Bacillus subtilis, thioredoxin reductase, prokaryotic expression, activity analysis

CLC Number:

Q939.9

CHEN Hong-na, LI Jian-wen, JIANG Ao, SUN Wen-xiu. Prokaryotic expression and activity analysis of thioredoxin reductase from Bacillus subtilis[J]. , 2016, 28(12): 2054-2059.

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Prokaryotic expression and activity analysis of thioredoxin reductase from Bacillus subtilis

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