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丙酸丙酸杆菌钠离子依赖性ATP合酶F0部分编码基因在大肠杆菌中的克隆、测序及体内表达

Cloning, sequencing and in vivo expression of genes encoding the F0 part of the sodium-ion-dependent ATP synthase of Propionigenium modestum in Escherichia coli.

作者信息

Kaim G, Ludwig W, Dimroth P, Schleifer K H

机构信息

Lehrstuhl für Mikrobiologie, Technische Universität München, Federal Republic of Germany.

出版信息

Eur J Biochem. 1992 Jul 15;207(2):463-70. doi: 10.1111/j.1432-1033.1992.tb17072.x.

Abstract

A DNA fragment containing the genes encoding subunits of the F0 part of the sodium-translocating ATPase of Propionigenium modestum was cloned in Escherichia coli and sequenced. The predicted amino acid sequences of subunits a, b and c of the P. modestum ATPase were compared with those of the corresponding subunits of proton-translocating ATPases from other bacteria and chloroplasts. Deletion mutants of E. coli, lacking different genes for ATPase subunits, were transformed with a recombinant plasmid, containing the genes for the subunits a, c, b, delta and part of alpha of the ATPase of P. modestum. Functionally reconstituted ATPase activity could be demonstrated for the transformants. The identity of the vector containing P. modestum genes was verified by restriction analysis of plasmid DNA.

摘要

将含有丙酸栖热菌钠转运ATP酶F0部分亚基编码基因的DNA片段克隆到大肠杆菌中并进行测序。将丙酸栖热菌ATP酶a、b和c亚基的预测氨基酸序列与其他细菌和叶绿体质子转运ATP酶相应亚基的序列进行了比较。用含有丙酸栖热菌ATP酶a、c、b、δ和部分α亚基基因的重组质粒转化缺乏不同ATP酶亚基基因的大肠杆菌缺失突变体。可证明转化体具有功能重构的ATP酶活性。通过对质粒DNA的限制性分析验证了含有丙酸栖热菌基因的载体的同一性。

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