CLONING, SEQUENCING AND DELETION FROM THE CHROMOSOME OF THE GENE ENCODING SUBUNIT-1 OF THE AA3-TYPE CYTOCHROME-C-OXIDASE OF RHODOBACTER-SPHAEROIDES

被引:68
作者
SHAPLEIGH, JP [1 ]
GENNIS, RB [1 ]
机构
[1] UNIV ILLINOIS,SCH CHEM SCI,URBANA,IL 61801
关键词
D O I
10.1111/j.1365-2958.1992.tb01511.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The ctaD gene encoding subunit I of the aa3-type cytochrome c oxidase from Rhodobacter sphaeroides has been cloned. The gene encodes a polypeptide of 565 residues which is highly homologous to the sequences of subunit I from other prokaryotic and eukaryotic sources, e.g. 51% identity with that from bovine, and 75% identity with that from Paracoccus denitrificans. The ctaD gene was deleted from the chromosome of R. sphaeroides, resulting in a strain that spectroscopically lacks cytochrome a. This strain maintains about 50% of the cytochrome c oxidase activity of the wild-type strain owing to the presence of an alternate o-type cytochrome c oxidase. The aa3-type oxidase was restored by complementing the chromosomal deletion with a plasmid-borne copy of the ctaD gene. This system is well suited for site-directed mutagenesis probing of the structure and function of cytochrome c oxidase.
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收藏
页码:635 / 642
页数:8
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