Glyoxylate bypass operon of Escherichia coli: cloning and determination of the functional map.

T. Chung, D. J. Klumpp, D. C. LaPorte

Research output: Contribution to journalArticle

57 Scopus citations

Abstract

In Escherichia coli, a single operon encodes the metabolic and regulatory enzymes of the glyoxylate bypass. The metabolic enzymes, isocitrate lyase and malate synthase, are expressed from aceA and aceB, and the regulatory enzyme, isocitrate dehydrogenase kinase/phosphatase, is expressed from aceK. We cloned this operon and determined its functional map by deletion analysis. The order of the genes in this operon is aceB-aceA-aceK, with aceB proximal to the promoter, consistent with the results of previous experiments using genetic techniques. The promoter was identified by S1 nuclease mapping, and its nucleotide sequence was determined. Isocitrate lyase and malate synthase were readily identified by autoradiography after the products of the operon clone were labeled by the maxicell procedure and then resolved by electrophoresis. In contrast, isocitrate dehydrogenase kinase/phosphatase, expressed from the same plasmid, was undetectable. This observation is consistent with a striking downshift in expression between aceA and aceK.

Original languageEnglish (US)
Pages (from-to)386-392
Number of pages7
JournalJournal of bacteriology
Volume170
Issue number1
DOIs
StatePublished - Jan 1988

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