Cloning, Expression and Purification of Wheat Acetyl-CoA Carboxylases CT Domain in E. coil  

作　　者：WANG Rui-jian YANG Xue-ying ZHENG Liang-yu YANG Ye GAO Gui CAO Shu-gui 

机构地区：[1]Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun 130021, P. R. China [2]College of Forestry, Beihua University, Jilin 132013, P. R. China

出　　处：《Chemical Research in Chinese Universities》2008年第6期752-755,共4页高等学校化学研究（英文版）

基　　金：Supported by the National Natural Science Foundation of China(Nos. 20432010, 20672045 and 30570405)

摘　　要：The entire gene of carboxyltransferase（CT） domain of acetyl-CoA carboxylase（ACCase） from Chinese Spring wheat（CSW） plastid was cloned firstly, and the 2.3 kb gene was inserted into PET28a^＋ vector and expressed in E. coil in a soluble state. The （His）6 fusion protein was identified by SDS-PAGE and Western blot. The recombinant protein was purified by affinity chromatography, and the calculated molecular mass（Mr） was 88000. The results of the sequence analysis indicate that the cloned gene（GeneBank accession No. EU124675） was a supplement and revision of the reported ACCase CT partial cDNA from Chinese Spring wheat plastid. The recombinant protein will be significant for us to investigate the recognizing mechanism between ACCase and herbicides, and further to screen new herbicides.The entire gene of carboxyltransferase（CT） domain of acetyl-CoA carboxylase（ACCase） from Chinese Spring wheat（CSW） plastid was cloned firstly, and the 2.3 kb gene was inserted into PET28a^＋ vector and expressed in E. coil in a soluble state. The （His）6 fusion protein was identified by SDS-PAGE and Western blot. The recombinant protein was purified by affinity chromatography, and the calculated molecular mass（Mr） was 88000. The results of the sequence analysis indicate that the cloned gene（GeneBank accession No. EU124675） was a supplement and revision of the reported ACCase CT partial cDNA from Chinese Spring wheat plastid. The recombinant protein will be significant for us to investigate the recognizing mechanism between ACCase and herbicides, and further to screen new herbicides.

关 键 词：Chinese Spring wheat Acetyl-CoA carboxylase Carboxyltransferase（CT） domain CLONING Expression PURIFICATION 

分 类 号：S512[农业科学—作物学]