PMID:3917997
| Citation |
Calhoun, DH, Wallen, JW, Traub, L, Gray, JE and Kung, HF (1985) Internal promoter in the ilvGEDA transcription unit of Escherichia coli K-12. J. Bacteriol. 161:128-32 |
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| Abstract |
Segments of the ilvGEDA transcription unit have been cloned into the promoter tester plasmid pMC81. This vector contains cloning sites situated upstream of the lacZ gene coding for beta-galactosidase. Using this method we have quantitatively evaluated in vivo (i) the activity of previously described promoter, pG, preceding ilvG; (ii) the relative activity of pE promoter, previously postulated to be located between ilvG and ilvE; and (iii) the effect of the frameshift site present in the wild-type ilvG gene by comparison with mutant derivatives lacking this frameshift site. Isogenic derivatives of strain MC1000 were constructed by transduction with phage P1 grown on rho-120, delta(ilvGEDA), delta(ilvED), and ilvA538 hosts. The potential effects of these alleles that were previously postulated to affect ilvGEDA expression were assessed in vivo by monitoring beta-galactosidase production directed by ilv DNA fragments. Cloned ilv segments were also tested for activity in vitro with a DNA-directed coupled transcription and translation system. The production in vitro of ilv-directed ilv gene expression and beta-galactosidase expression with ara-ilv-lac fusions paralleled the in vivo activity. |
| Links | |
| Keywords |
Bacteriophages/genetics; Cloning, Molecular; DNA, Bacterial/genetics; Escherichia coli/genetics; Mutation; Operon; Plasmids; Protein Biosynthesis; Transcription, Genetic; Transduction, Genetic; beta-Galactosidase/genetics |
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