Internal promoter in the ilvGEDA transcription unit of Escherichia coli K-12
| Autor: | L. Traub, J. E. Gray, D. H. Calhoun, J. W. Wallen, Hsiang-Fu Kung |
|---|---|
| Rok vydání: | 1985 |
| Předmět: |
DNA
Bacterial Transcription Genetic Operon Mutant Biology Molecular cloning beta-Galactosidase Microbiology Molecular biology Frameshift mutation Plasmid Transduction Genetic Transcription (biology) Protein Biosynthesis Mutation Gene expression Escherichia coli Bacteriophages Cloning Molecular Molecular Biology Gene Plasmids Research Article |
| Zdroj: | Journal of Bacteriology. 161:128-132 |
| ISSN: | 1098-5530 0021-9193 |
| DOI: | 10.1128/jb.161.1.128-132.1985 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|