PMID:6178738
| Citation |
Brosius, J, Cate, RL and Perlmutter, AP (1982) Precise location of two promoters for the beta-lactamase gene of pBR322. S1 mapping of ribonucleic acid isolated from Escherichia coli or synthesized in vitro. J. Biol. Chem. 257:9205-10 |
|---|---|
| Abstract |
We identified two promoters for the beta-lactamase gene of plasmid pBR322. RNA isolated from bacteria containing pBR322 or RNA transcribed in vitro on pBR322 templates was hybridized to 5' end-labeled single-stranded plasmid probes (Berk, A. J., and Sharp, P. A. (1977) Cell 12, 721-732). Electrophoretic analysis of the nuclease S1 digestion products next to Maxam-Gilbert sequencing ladders closely defines the transcriptional initiation points. The natural promoter lies near the coding sequence of the beta-lactamase gene, initiating transcription at -35 bases before the ATG initiation codon, while a second promoter initiates at positions -244 and/or -245 (on the opposite side of the Eco RI site). This promoter overlaps the promoter transcribing in the opposite direction toward the tetracycline gene(s) and starts in the -10 region of that promoter. S1 mapping of procaryotic mRNA, transcribed in vivo, allows both an accurate identification of promoters and the analysis of their transcriptional regulation. |
| Links | |
| Keywords |
Base Sequence; Electrophoresis, Polyacrylamide Gel; Endonucleases/metabolism; Escherichia coli/genetics; Genes, Bacterial; Operon; RNA, Bacterial/analysis; Single-Strand Specific DNA and RNA Endonucleases; Templates, Genetic; Transcription, Genetic; beta-Lactamases/genetics |
| edit table |
Significance
You can help EcoliWiki by summarizing why this paper is useful
Useful Materials and Methods
You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.
Annotations
<annotationlinks/>
EcoliWiki Links
Add links to pages that link here (e.g. gene, product, method pages)
References
See Help:References for how to manage references in EcoliWiki.