Cloning，Prokaryotic Expression of Sorghum bicolor SbGABA-Ts

doi:10.13560/j.cnki.biotech.bull.1985.2015.05.015

Abstract

Abstract: GABA is a ubiquitous four-carbon，non-protein amino acid，and has been associated with growth and development，signaling transduction，and stress response in plants. GABA transaminase（GABA-T），the enzyme responsible for the catabolism of GABA，has not been fully explored，especially in several important crops. Two putative GABA transaminase genes（GABA-T）from Sorghum bicolor were obtained based on homology analysis with the identified GABA-Ts of other plants and RT-PCR. Subsequently，the two SbGABA-T genes and corresponding N-terminal truncated SbGABA-Ts were inserted into pET28a（+）vector and individually imported into E. coli strain BL21（pLysS，DE3）. Percentage improvement of soluble recombinant proteins were showed when removing the targeting peptide sequence of SbGABA-Ts. The fusion proteins were expressed partially in soluble form after incubating for 18 h at 16℃ by adding 1 mmol/L IPTG，and purified through nickel-affinity chromatography column.

Key words: Sorghum bicolor GABA-T, prokaryotic expression, purification, signal peptide

Yang Zewei, Wang Longhai, Zhu Li, Wang Hai, Huang Dafang, Lang Zhihong. Cloning，Prokaryotic Expression of Sorghum bicolor SbGABA-Ts[J]. Biotechnology Bulletin, 2015, 31(5): 93-99.

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