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Procedure for the rapid, large-scale purification of Escherichia coli DNA-dependent RNA polymerase involving polymin P precipitation and DNA-cellulose chromatography
Abstract
An improved method is described for the purification of the DNA-dependent RNA polymerase [ribonucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6] from Escherichia coli. The method involves lysozyme-sodium deoxycholate lysis, low-speed centrifugation, precipitation with Polymin P, elution from the Polymin P precipitate, ammonium sulfate precipitation, and chromatography on DNA-cellulose and Bio-Gel A 5m. RNA polymerase is purified to electrophoretic homogeneity in 2 days with a recovery of 45%, resulting in a yield of 250 mg of holoenzyme from 500 g of cells.
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Cites
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
LAEMMLI, U. K. (1970) · Nature · doi:10.1038/227680a0 · Cited by 249,961
Cited by
Kruger, K., Grabowski, P. J., Zaug, A. J., Sands, J., Gottschling, D. E., & Cech, T. R. (1982) · Cell · doi:10.1016/0092-8674(82)90414-7 · Cited by 2,331
Promoters largely determine the efficiency of repressor action.
Lanzer, M., & Bujard, H. (1988) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.85.23.8973 · Cited by 351
Geier, G. E., & Modrich, P. (1979) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)34217-5 · Cited by 332
Identification and characterization of E. coli CRISPR‐cas promoters and their silencing by H‐NS
Pul, Ü., Wurm, R., Arslan, Z., Geißen, R., Hofmann, N., & Wagner, R. (2010) · Molecular Microbiology · doi:10.1111/j.1365-2958.2010.07073.x · Cited by 282
Metadata
- Authors
- Burgess, R. R., & Jendrisak, J. J.
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1975-10-01
- Volume
- 14
- Issue
- 21
- Pages
- 4634-4638
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2030551247
- Cited by
- 1,144
- References
- 1
- FWCI
- 29.06
- Type
- journal article
- Language
- en
- Open access
- closed
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- Burgess, R. R., & Jendrisak, J. J. (1975). Procedure for the rapid, large-scale purification of Escherichia coli DNA-dependent RNA polymerase involving polymin P precipitation and DNA-cellulose chromatography. Biochemistry. https://doi.org/10.1021/bi00692a011
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
