苜蓿中华根瘤菌Rm1021中gtrA基因的克隆、表达及纯化

Cloning,Expression and Purification of gtrA Gene in Sinorhizobium meliloti Rm 1021

对苜蓿中华根瘤菌中GntR家族转录因子基因gtrA进行了表达和纯化,以期进一步研究其功能。以苜蓿根瘤菌Rm1021的基因组为模板,PCR扩增出gtrA基因,并克隆到表达载体PET-28b(+)上。重组质粒经酶切和测序证实正确,然后转化大肠杆菌BL21。‘SDS-PAGE显示经IPTG诱导后能得到和理论值大小一致的蛋白条带,通过进一步纯化得到gtrA的表达蛋白。

In this paper, the GntR- like tanscription gene gtrA from Sinorhizobium meliloti was expressed and purified to further research its function. The gtrA gene was amplified by PCR from Sinorhizobium meliloti Rm1021 genomic DNA, and then this gene was cloned on PET - 28b （ ＋ ） expressing vector. The recombinant plasmid was confirmed by digestion and sequence, then transformed into E. coli BL21. The results of SDS - PAGE showed that E. coli BL21 contained the recombinant plasmid expressing the fight molecular weight protein after inducing by IPTG, then the gtrA protein of gtrA gene was obtanined by purification.