长喙田菁植物螯合肽合成酶PCS1的原核表达及纯化

为了获得纯化的长喙田菁(Sesbania rostrata)植物螯合肽合成酶PCS1,本研究以原核表达载体pMAL-c2x为基础,构建了含有SrPCS1开放阅读框序列的原核表达载体pAM56,并将其转化大肠杆菌表达菌株BL21(DE3),对融合蛋白的表达进行了优化,通过Western blotting鉴定融合蛋白,且用...

Prokaryotic Expression and Purification of Phytochelatin Synthasel in Escherichia coli of Sesbania rostrata

In order to obtain purified Sesbania rostrata phytochelatin synthase1,the prokaryotic expressing vector pAM56 containing SrPCS1 open reading frame was constructed based on the vector pMAL-c2x and transferred into E.coli BL21(DE3) and induce the fusion protein MBP-SrPCS1.The fusion protein was then identified by western blotting and purified through Maltose Binding column.The results suggested that the soluble protein MBP-SrPCS1 was induced to express through 0.2 mmol/L of IPTG after 16 h at 15 ℃ and the purified MBP-SrPCS1 was obtained by affinity chromatography,which can lay a foundation for further study on the enzyme activity of SrPCS1 and the catalytic mechanism of PCS.