Abstract
Immobilized enzymes or nucleic acids are widely used to analyze association and dissociation reactions. A pseudo-rapid kinetic method for studying transcription was developed by using an immobilized template DNA packed in a minute column, in a micro-scale liquid chromatography system. This method, which has a dead-time of only several seconds, is quick enough to allow analysis of the release of product RNA in transcription by Escherichia coli RNA polymerase. The method could be applied to most enzymes that interact with DNA.
Full Text
The Full Text of this article is available as a PDF (59.8 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Carpousis A. J., Gralla J. D. Cycling of ribonucleic acid polymerase to produce oligonucleotides during initiation in vitro at the lac UV5 promoter. Biochemistry. 1980 Jul 8;19(14):3245–3253. doi: 10.1021/bi00555a023. [DOI] [PubMed] [Google Scholar]
- Fujioka M., Hirata T., Shimamoto N. Requirement for the beta,gamma-pyrophosphate bond of ATP in a stage between transcription initiation and elongation by Escherichia coli RNA polymerase. Biochemistry. 1991 Feb 19;30(7):1801–1807. doi: 10.1021/bi00221a011. [DOI] [PubMed] [Google Scholar]
- Kubori T., Shimamoto N. A branched pathway in the early stage of transcription by Escherichia coli RNA polymerase. J Mol Biol. 1996 Mar 1;256(3):449–457. doi: 10.1006/jmbi.1996.0100. [DOI] [PubMed] [Google Scholar]
- Langer P. R., Waldrop A. A., Ward D. C. Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6633–6637. doi: 10.1073/pnas.78.11.6633. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levin J. R., Krummel B., Chamberlin M. J. Isolation and properties of transcribing ternary complexes of Escherichia coli RNA polymerase positioned at a single template base. J Mol Biol. 1987 Jul 5;196(1):85–100. doi: 10.1016/0022-2836(87)90512-2. [DOI] [PubMed] [Google Scholar]
- Munson L. M., Reznikoff W. S. Abortive initiation and long ribonucleic acid synthesis. Biochemistry. 1981 Apr 14;20(8):2081–2085. doi: 10.1021/bi00511a003. [DOI] [PubMed] [Google Scholar]
- Zawel L., Kumar K. P., Reinberg D. Recycling of the general transcription factors during RNA polymerase II transcription. Genes Dev. 1995 Jun 15;9(12):1479–1490. doi: 10.1101/gad.9.12.1479. [DOI] [PubMed] [Google Scholar]