Molecular Biology of the Cytochrome O Terminal Oxidase of Escherichia Coli (Cloning, Sequencing, Dna)
Au, Chi-Tak Doug
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https://hdl.handle.net/2142/70358
Description
Title
Molecular Biology of the Cytochrome O Terminal Oxidase of Escherichia Coli (Cloning, Sequencing, Dna)
Author(s)
Au, Chi-Tak Doug
Issue Date
1987
Department of Study
Chemistry
Discipline
Chemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Chemistry, Biochemistry
Abstract
My thesis research has dealt mainly with a genetic approach to studying the cytochrome o terminal oxidase complex of Escherichia coli. Respiratory-deficient mutants of E. coli have been isolated which are unable to grow aerobically on non-fermentable substrates such as succinate and lactate. Spectroscopic and immunological studies showed that these mutants lack the cytochrome o terminal oxidase of the high aeration branch of the respiratory chain. These strains carry mutations in a locus designated cyo which maps at min 10.2 on the E. coli linkage map. The structural genes encoding the cytochrome o oxidase complex have been cloned into the multicopy plasmid pBR322. Strains carrying the cyo plasmids produce 5 to 10 times more cytochrome o oxidase than control strains. The gene products encoded by the cyo plasmids can be immunoprecipitated with monospecific antibodies raised against cytochrome o. The cyo gene locus is located in a 5.5-kb chromosomal DNA segment. The nucleotide sequence of a portion of the cyo gene locus has been determined by Sanger's dideoxynucleotide sequencing method. Further genetic manipulations and a complete DNA sequence analysis of the cyo gene locus will provide valuable information on the protein structure-function relationships.
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