PMID:6308128
| Citation |
Guest, JR, Roberts, RE and Stephens, PE (1983) Hybrid plasmids containing the pyruvate dehydrogenase complex genes and gene-DNA relationships in the 2 to 3 minute region of the Escherichia coli chromosome. J. Gen. Microbiol. 129:671-80 |
|---|---|
| Abstract |
A sample of colonies from the Clarke-Carbon ColE1-Escherichia coli DNA plasmid gene bank was screened by conjugation for complementation of the lipoamide dehydrogenase lesion of a deletion strain lacking all components of the pyruvate dehydrogenase complex, delta (aroP aceE aceF lpd). Two ColE1-lpd+ hybrid plasmids were identified: pGS2 (ColE1-ace lpd+; 24 kb) and pGS5 (ColE1-lpd+; 14 kb). Enzymological studies confirmed that pGS2 expressed all the activities of the pyruvate dehydrogenase complex, whereas pGS5 expressed the lipoamide dehydrogenase and acetyltransferase activities (the latter from a ColE1 promoter). These and other plasmids were used to construct a 47-site (15 enzymes) restriction map for a 24.2 kb segment of bacterial DNA in the nadC-lpd region. A further 13 sites (six enzymes) were defined in a 5.4 kb sub-segment containing the lpd gene. lambda phage derivatives containing specific fragments were constructed and used in transduction studies which located the ace and lpd genes in a 7.78 kb sub-segment flanked by AccI and NruI sites. |
| Links | |
| Keywords |
Bacteriophage lambda/genetics; Chromosome Mapping; Chromosomes, Bacterial; Conjugation, Genetic; DNA Restriction Enzymes; DNA, Bacterial/genetics; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli/ultrastructure; Genes, Bacterial; Genotype; Hybridization, Genetic; Plasmids; Pyruvate Dehydrogenase Complex/genetics; Transduction, Genetic |
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