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Title:Molecular genetic studies of the cytochrome o terminal oxidase complex in Escherichia coli
Author(s):Chepuri, Visala
Doctoral Committee Chair(s):Gennis, Robert B.
Department / Program:Biochemistry
Degree Granting Institution:University of Illinois at Urbana-Champaign
Subject(s):Biology, Molecular
Biology, Genetics
Chemistry, Biochemistry
Abstract:The cytochrome $o$ terminal oxidase complex is a component of aerobic respiratory chain of Escherichia coli. This enzyme catalyzes the oxidation of ubiquinol-8 to ubiquinone-8 within the cytoplasmic membrane and the concomitant reduction of O$\sb2$ to H$\sb2$O. Cytochrome $o$ oxidase has been purified and shown to contain four subunits on SDS-PAGE gels. The cloning of the $cyo$ operon makes it possible to address a number of problems concerning the structure and function of the cytochrome $o$ oxidase complex. Detailed analysis of this operon will enable us to identify the subunit compositions, the locations of the hemes, etc.
The thesis research described here has mainly dealt with the structure-function relationship of the cytochrome $o$ oxidase complex. Molecular biological, genetic and some biochemical approaches were used to study the following aspects of cytochrome $o$ oxidase complex: (i) sequence of the $cyo$ operon to determine the primary structure and subunit composition of the cytochrome $o$ oxidase complex, (ii) sequence portions of the polypeptides of cytochrome $o$ complex to correlate with the corresponding ORF's in the $cyo$ operon and to confirm the accuracy of the ORF's deduced by DNA sequence analysis, (iii) use the method of gene fusions to determine the topology of all the subunits of the cytochrome $o$ oxidase complex, (iv) construct a cyo-lacZ fusion and transfer it to the chromosome in order to study regulation in single copy, in this case by the $arc$ system, (v) localize both the cytochromes of the cytochrome $o$ oxidase complex by deletion analysis, and (vi) establish conditions in order to isolate HQNO resistant mutants of the cytochrome $o$ oxidase complex.
Issue Date:1990
Rights Information:Copyright 1990 Chepuri, Visala
Date Available in IDEALS:2011-05-07
Identifier in Online Catalog:AAI9026156
OCLC Identifier:(UMI)AAI9026156

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