PMID:6296045
| Citation |
Guest, JR and Roberts, RE (1983) Cloning, mapping, and expression of the fumarase gene of Escherichia coli K-12. J. Bacteriol. 153:588-96 |
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| Abstract |
Two classes of fumarase-transducing phages, lambda fumA and lambda fumB, were isolated from populations of recombinant phages containing HindIII fragments of Escherichia coli DNA; they were isolated by virtue of their ability to complement the metabolic lesion of a fumarase-negative mutant. The strongly complementing lambda fumA phages contained a 6.2-kilobase HindIII fragment encoding: the fumA gene, located at 35.5 min in the E. coli linkage map and expressing the major fumarase activity; the mannosephosphate isomerase gene, manA; and an unidentified gene, g48. The three genes were located relative to the restriction map of the cloned fragment and the genetic linkage map (terC-g48-fumA-manA-uidAoR), their transcription polarities were defined as anticlockwise in the chromosome, and the molecular weights of the corresponding gene products were established: fumA, 61,500; manA, 42,000; g48, 48,000. Organisms containing the fumA gene sub-cloned in multicopy plasmids overproduced fumarase up to 50-fold. The weakly complementing class of transducing phages, lambda fumB, contained several genes in an 8.2-kilobase HindIII fragment, including one (fumB) that determines a minor fumarase activity. Complementation by fumB was only observed in high-copy situations such as transduction plaques and in strains containing a multicopy plasmid in which 40% of normal fumarase activity was detected. The basis for the complementation by fumB was not defined. |
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| Keywords |
Bacteriophage lambda/genetics; Chromosome Mapping; Chromosomes, Bacterial; Cloning, Molecular; DNA Restriction Enzymes; Escherichia coli/enzymology; Escherichia coli/genetics; Fumarate Hydratase/genetics; Gene Expression; Genes; Genes, Bacterial; Lysogeny; Mannose-6-Phosphate Isomerase/genetics; Plasmids; Transduction, Genetic |
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