期刊论文

中文

中国农业科学

0578-1752

2009

42

7

2297-2304

06FJ3180:湖南省自然科学基金 07JJ5040:湖南省科技计划

本校为第一机构

[目的]揭示棉田杂草薤白(Allium macrostemon Bunge)抗草甘膦的分子机理,开发利用其草甘膦抗性特性.[方法]运用RT-PCR结合RACE技术,分离克隆了薤白中草甘膦作用的靶酶EPSP合成酶(EPSPs)基因cDNA,并将其构建成原核表达载体在大肠杆菌中诱导表达和鉴定其抗性.[结果]薤白EPSP合成酶基因cDNA序列全长1821 bp,编码一段522个氨基酸的推导蛋白质.经BLAST及蛋白质结构预测,蛋白具有EPSPs的特征序列,并与已报道的EPSPs序列有高同源性,确认克隆的cDNA序列即是薤白EPSPs基因序列,命名为EPSPsA.将该cDNA与原核表达载体pRSET-A重组后,构建成重组表达质粒pRSET-A-EPSPsA,并转化至大肠杆菌BL21(DE3),用IPTG诱导...

[ Objective ] Allium macrostemon Bunge is a weed plant and it is very hard to control in cotton field because of its glyphosate resistance. To reveal its mechanism of glyphosate resistance can lead to a molecular understanding and utilization. [Method] The cDNA of EPSPs gene that is responsible to glyphosate resistance were amplified and cloned by RT-PCR and RACE. The glyphosate resistance of the protein is tested by expression of the EPSPs cDNA in E. coli. [Result] The cloned full-length cDNA is 1 821 bp with an open reading frame of 1 569 bp....