生物技术通报2015，Vol.31Issue(5)：93-99,7.DOI:10.13560/j.cnki.biotech.bull.1985.2015.05.015
高粱SbGABA-Ts基因的克隆、原核表达及纯化
Cloning,Prokaryotic Expression of Sorghum bicolorSbGABA-Ts
摘要
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-Tgenes and corresponding N-terminal truncated SbGABA-Ts were inserted into pET28a(+)vector and individually imported intoE. 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.关键词
高粱GABA-T/原核表达/纯化/信号肽Key words
Sorghum bicolorGABA-T/prokaryotic expression/purification/signal peptide引用本文复制引用
杨泽伟,王龙海,朱莉,汪海,黄大昉,郎志宏..高粱SbGABA-Ts基因的克隆、原核表达及纯化[J].生物技术通报,2015,31(5):93-99,7.基金项目
国家自然科学基金项目(31271790,31471558) （31271790,31471558）
