摘要
为研究高抗草甘膦N-乙酰转移酶(glyphosate N-acetyltransferase,GAT)的草甘膦抗性相关位点,以高抗草甘膦的gat1基因和不具草甘膦抗性的gat2基因为出发基因,通过一轮双基因DNA shuffling,获得了抗性较野生型gat1下降的突变体14个:10个单位点突变,3个双位点突变,1个3位点突变。其中,MTGAT3(H41Q)、MTGAT5(S52P)、MTGAT6(I53M)、MTGAT7(F56L)、MTGAT8(R82G)、MTGAT11(I19T/P22L)和MTGAT14(Y70H/L98P/K119E)7个突变体的草甘膦抗性几乎丧失。本工作为深入研究GAT1蛋白分子结构与功能之间的关系奠定了基础。
To study on some sites related to glyphosate-tolerance in glyphosate N-acetyhransferase (GAT) with high glyphosate-tolerance, the high glyphosate-tolerant gatl gene and the gat2 gene nearly without glyphosate-tolerance were used to directed evolution by DNA shuffling. As a result, fourteen gatl mutants containing lower glyphosatetolerance than wild-type gatl were obtained by selection with different concentrations of glyphosate (5 mmol · L^-1 to 50 mmol · L^- 1 ). Ten of them were single site mutated, three ones were double sites mutated, while one was three sites mutated. Among them, the glyphosate-tolerance of mutants MTGAT3 ( H41Q ), MTGAT5 ( S52P ), MTGAT6 (I53M) ,MTGAT7 (F56L) ,MTGAT8 (R82G) ,MTGAT11 (I19T/P22L) and MTGAT14(Y70H/L98P/K119E) were almost lost. This research work has laid a foundation for further studying the relationship between GAT1 protein structure and its function.
出处
《中国农业科技导报》
CAS
CSCD
2007年第5期110-114,共5页
Journal of Agricultural Science and Technology
基金
国家自然科学基金项目"苯胺双加氧酶诱导型启动子和调控蛋白互作的分子机理"(30470047)资助