Method for enhancement of plant redox-related protein expression and its application for in vitro reduction of chloroplastic thioredoxins
Method for enhancement of plant redox-related protein expression and its application for in vitro reduction of chloroplastic thioredoxins
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DOI：
10.1016/j.pep.2014.07.001
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发表时间：
2014-09-01
影响因子：
1.6
通讯作者：
Okegawa, Yuki
中科院分区：
文献类型：
--
作者：
Motohashi, Ken;Okegawa, Yuki
使用翻译起始密码子下游的遗传密码子替换来过度表达植物氧化还原相关蛋白。该方法显着提高了大肠杆菌中拟南芥叶绿体硫氧还蛋白和胞质烟酰胺腺嘌呤二核苷酸磷酸 (NADPH) 依赖性硫氧还蛋白还原酶 (E.C. 1.8.1.9) 的重组蛋白表达水平。使用这些蛋白质，以 NADPH 依赖性方式重建体外叶绿体硫氧还蛋白还原系统。该系统可以将五类叶绿体拟南芥硫氧还蛋白和两种叶绿体菠菜硫氧还蛋白转化为其还原形式，不依赖于二硫苏糖醇和光合电子传递系统。 (C) 2014 Elsevier Inc. 保留所有权利。
Plant redox-related proteins were overexpressed using a genetic codon substitution downstream of the translation initiation codon. This method significantly improved recombinant protein expression levels of Arabidopsis chloroplastic thioredoxins and cytosolic nicotinamide adenine dinucleotide phosphate (NADPH)-dependent thioredoxin reductase (E.C. 1.8.1.9) in Escherichia coli. Using these proteins, the in vitro chloroplastic thioredoxins-reduction system was reconstituted in an NADPH-dependent manner. This system could convert the five classes of chloroplastic Arabidopsis thioredoxins and two chloroplastic Spinach thioredoxins to their reduced forms, independent of dithiothreitol and the photosynthetic electron transport system. (C) 2014 Elsevier Inc. All rights reserved.