Rui-jian WANG , Xue-ying YANG , Liang-yu ZHENG , Ye YANG , Gui GAO , Shu-gui CAO
{"title":"小麦乙酰辅酶a羧化酶CT结构域的克隆、表达和纯化","authors":"Rui-jian WANG , Xue-ying YANG , Liang-yu ZHENG , Ye YANG , Gui GAO , Shu-gui CAO","doi":"10.1016/S1005-9040(09)60021-0","DOIUrl":null,"url":null,"abstract":"<div><p>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<sup>+</sup> vector and expressed in <em>E. coil</em> in a soluble state. The (His)<sub>6</sub> fusion protein was identified by SDS-PAGE and Western blot. The recombinant protein was purified by affinity chromatography, and the calculated molecular mass(<em>M</em><sub>r</sub>) 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.</p></div>","PeriodicalId":9785,"journal":{"name":"Chemical Research in Chinese Universities","volume":"24 6","pages":"Pages 752-755"},"PeriodicalIF":3.1000,"publicationDate":"2008-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1005-9040(09)60021-0","citationCount":"0","resultStr":"{\"title\":\"Cloning, Expression and Purification of Wheat Acetyl-CoA Carboxylases CT Domain in E. coil\",\"authors\":\"Rui-jian WANG , Xue-ying YANG , Liang-yu ZHENG , Ye YANG , Gui GAO , Shu-gui CAO\",\"doi\":\"10.1016/S1005-9040(09)60021-0\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>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<sup>+</sup> vector and expressed in <em>E. coil</em> in a soluble state. The (His)<sub>6</sub> fusion protein was identified by SDS-PAGE and Western blot. The recombinant protein was purified by affinity chromatography, and the calculated molecular mass(<em>M</em><sub>r</sub>) 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.</p></div>\",\"PeriodicalId\":9785,\"journal\":{\"name\":\"Chemical Research in Chinese Universities\",\"volume\":\"24 6\",\"pages\":\"Pages 752-755\"},\"PeriodicalIF\":3.1000,\"publicationDate\":\"2008-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/S1005-9040(09)60021-0\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Chemical Research in Chinese Universities\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1005904009600210\",\"RegionNum\":4,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"CHEMISTRY, MULTIDISCIPLINARY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Chemical Research in Chinese Universities","FirstCategoryId":"92","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1005904009600210","RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
Cloning, Expression and Purification of Wheat Acetyl-CoA Carboxylases CT Domain in E. coil
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.
期刊介绍：
The journal publishes research articles, letters/communications and reviews written by faculty members, researchers and postgraduates in universities, colleges and research institutes all over China and overseas. It reports the latest and most creative results of important fundamental research in all aspects of chemistry and of developments with significant consequences across subdisciplines.
Main research areas include (but are not limited to):
Organic chemistry (synthesis, characterization, and application);
Inorganic chemistry (bio-inorganic chemistry, inorganic material chemistry);
Analytical chemistry (especially chemometrics and the application of instrumental analysis and spectroscopy);
Physical chemistry (mechanisms, catalysis, thermodynamics and dynamics);
Polymer chemistry and polymer physics (mechanisms, material, catalysis, thermodynamics and dynamics);
Quantum chemistry (quantum mechanical theory, quantum partition function, quantum statistical mechanics);
Biochemistry;
Biochemical engineering;
Medicinal chemistry;
Nanoscience (nanochemistry, nanomaterials).
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