长喙田菁植物螯合肽合成酶PCS1的原核表达及纯化  

Prokaryotic Expression and Purification of Phytochelatin Synthase1 in Escherichia coli of Sesbania rostrata

作　　者：李安明[1] 邓青云[1] 李德华[1] 

机构地区：[1]孝感学院生命科学技术学院,湖北孝感432000

出　　处：《四川农业大学学报》2011年第2期213-217,共5页Journal of Sichuan Agricultural University

基　　金：湖北省重点(培育)学科:农业资源利用(0903);湖北省自然科学基金项目(2005ABA083)

摘　　要：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,且用麦芽糖亲和柱对MBP-SrPCS1融合蛋白进行了纯化。结果表明:在15℃下,经0.2 mmol/L IPTG诱导16 h表达出可溶性的MBP-SrPCS1融合蛋白,并通过亲和层析得到了纯化的MBP-SrPCS1融合蛋白,为进一步研究SrPCS1的活性及PCS的催化机制奠定了基础。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.

关 键 词：植物螯肽合成酶 SrPCS1 原核表达 纯化 

分 类 号：Q78[生物学—分子生物学]