Cloning，Subcellular Localization and Prokaryotic Expression of Gene TsGPX3 from Thellungiella salsuginea

doi:10.13560/j.cnki.biotech.bull.1985.2016.04.014

Abstract

Abstract: A cDNA of TsGPX3（Thellungiella salsuginea GPX3）was isolated from T. salsuginea，and it contained a complete ORF with 591 bp encoding 196 amino acid residues. The three conservative domains of TsGPX3 protein in GPXs family was predicted by bioinformatics analysis. Phylogenetic analysis discovered that TsGPX3 was in high homology with RsGPX3 of Raphanus sativus，BnGPX3 of Brassica napus，and BoGPX3 of B. oleracea etc.，all belonging to Brassicaceae. The plant-expressed vector pRTL2-TsGPX3-GFP was transiently expressed by transforming the protoplasts of Arabidopsis thaliana via PEG method，and it was discovered that the TsGPX3 protein was mainly located in the plasma membrane. The constructed prokaryotic vector pET-TsGPX3 was transferred into Escherichia coli strain BL21（DE3）for induced expression under different concentration of IPTG，and a 27 kD recombinant protein was highly expressed after induced for 5 h at 37℃.

Key words: TsGPX3, Thellungiella salsuginea, subcellular localization, prokaryotic expression

WANG Ning, CHEN Jing，, GAO Fei ,LI Hua-yun ,SUI Xin, ZHOU Yi-jun. Cloning，Subcellular Localization and Prokaryotic Expression of Gene TsGPX3 from Thellungiella salsuginea[J]. Biotechnology Bulletin, 2016, 32(4): 110-115.

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