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玉米叶绿体atp B及atp E基因在大肠杆菌中的表达

王振国, 魏家绵, 沈允钢

中科院上海植物生理研究所. 上海 200032

摘要

构建了在大肠杆菌中能表达玉米叶绿体CF_(1)完整#beta#亚基、缺失N端23个氨酸残基的#beta#亚基以及完整#epsilon#亚基的表达质粒。SDS-聚丙烯酰胺凝胶电泳及Western blot实验表明, 在分别含这些表达质粒的细菌中#beta#、#epsilon#亚基蛋白的合 成量在诱导3h后即达到饱和, 其含量各占细菌体总蛋白的10％左右。这些表达产物在细 菌体中形成不溶性的包含体。可通过对菌体蛋白的分级分离, 将表达产物溶于5mol/L的尿素溶液中。

关键词： 叶绿体; atpB基因; atpE基因

Over-expression of maize chloroplast atp B and atp E genes in Escherichia coli

Wang Zhenguo, Wei Jiamian, Shen Yungang

Shanghai Institute of Plant Physiology, Chinese Academy of Sciences. Shanghai 200032

Abstract

We constructed expressioin plasmids for high-level expression in Escherichia coli) of entire #beta# subunit of CF_(1) of maize chloroplast, #beta# subunit from which had been deleted twenty-three amino acid residues in the N-terminal region and entire #epsilon# subunit. SDS-polyacrylamide gel electrophoresis and Western blot experiments showed that the degree of expression of these proteins in E. coli) cells containing the corresponding expression plasmids reached saturuation after induction for 3h, accounting for 10% of total cell proteins respectively These expression products formed insoluble inclusion bodies in E. coli) cells and can be solubilized in 5mol/L urea after fractionation of E. coli) cell proteinase K.

Key words: Dsoroplast;atpB gene;atpE gage

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引用本文：

王振国, 魏家绵, 沈允钢. 玉米叶绿体atp B及atp E基因在大肠杆菌中的表达[J]. 生理学报 1995; 47 (3): .

Wang Zhenguo, Wei Jiamian, Shen Yungang. Over-expression of maize chloroplast atp B and atp E genes in Escherichia coli. Acta Physiol Sin 1995; 47 (3): (in Chinese with English abstract).