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

IF 3.1 4区 化学 Q2 CHEMISTRY, MULTIDISCIPLINARY
Rui-jian WANG , Xue-ying YANG , Liang-yu ZHENG , Ye YANG , Gui GAO , Shu-gui CAO
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引用次数: 0

Abstract

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.

小麦乙酰辅酶a羧化酶CT结构域的克隆、表达和纯化
首先克隆了中国春小麦(CSW)质体乙酰辅酶a羧化酶(ACCase)羧化转移酶(CT)结构域的全基因,并将该基因插入PET28a+载体中,在E. coil中以可溶性状态表达。通过SDS-PAGE和Western blot对(His)6融合蛋白进行鉴定。重组蛋白经亲和层析纯化,计算分子量(Mr)为88000。序列分析结果表明,克隆的基因(GeneBank accession No. 5291)具有克隆性。EU124675)是对已报道的中国春小麦质体ACCase CT部分cDNA的补充和修订。该重组蛋白对研究ACCase与除草剂的识别机制以及筛选新的除草剂具有重要意义。
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来源期刊
CiteScore
5.30
自引率
6.50%
发文量
152
审稿时长
3.0 months
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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