用体外定向进化的方法对EPSPS合酶基因草苷膦抗性机制的研究  

The in vitro directed evolution of EPSPS gene to identify of base pairs related to resistant to glyphosate

作　　者：王劲[1] 徐婧[2] 金丹[3] 杜世章[1] 

机构地区：[1]绵阳师范学院基因研究中心,绵阳621000 [2]东北农业大学资源与环境学院,哈尔滨150030 [3]四川大学生命科学学院,成都610064

出　　处：《四川大学学报（自然科学版）》2007年第1期155-159,共5页Journal of Sichuan University(Natural Science Edition)

基　　金：绵阳师范学院科研基金(MA2003006);四川省教育厅自然科学基金(2003B044);绵阳市应用技术研发基金(2004BS007)

摘　　要：为了寻找EPSP合酶中某些与草苷膦抗性相关的位点,利用体外定向进化技术,对可变盐单胞菌HT7的EPSP合酶基因,荧光假单胞菌G2的EPSP合酶基因以及按植物偏爱密码子合成G2的EPSP合酶基因,进行DNA shuffling,通过EPSP合酶缺陷型菌株E.coilER2799的功能互补筛选,获得了7个草苷膦抗性有较大变化的EPSP合酶突变体.It is the report of directed evolution of EPSPS genes using DNA shuffling in order to find some glyphosate-tolerant sites. The novel EPSPs gene from Halomonas Yariabilis strain HT7, EPSPs gene from Psedomonas flouorescens Strain G2 and the MG2 gene that modify the codon selection for all the amino acids to be consistent with normal usage in plant genes are used to directed evolution. Seven EPSPs mutants which conferring glyphosate-tolerance are screened using complement of E. coli ER2799.

关 键 词：体外定向进化 草苷膦 EPSP合酶 

分 类 号：Q78[生物学—分子生物学]