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Purification of Escherichia coli pulse-labeled RNA by benzoylated DEAE-cellulose chromatography

Sedat, John and Lyon, Alexander and Sinsheimer, Robert L. (1969) Purification of Escherichia coli pulse-labeled RNA by benzoylated DEAE-cellulose chromatography. Journal of Molecular Biology, 44 (3). pp. 415-434. ISSN 0022-2836. https://resolver.caltech.edu/CaltechAUTHORS:20160802-121819629

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Abstract

The rapidly-labeled RNA fraction of Escherichia coli has been purified approximately 15-fold on benzoylated DEAE-cellulose columns. It is metabolically unstable (as shown by a pulse/chase experiment), and is considered to represent messenger RNA. The yield of pulse-labeled RNA is about 70% and comprises 4 to 5% of the RNA of the cell. The true size distribution of this RNA, determined by sedimentation in a denaturing solvent (99% dimethyl sulfoxide), does not change during purification. This result indicates that neither degradation nor selection for molecules of a particular size has occurred. Upon sedimentation of the final preparation in dimethyl sulfoxide, the distribution of pulse label is the same as that of RNA mass, indicating nearly complete separation from longer-lived RNA components.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/0022-2836(69)90370-2DOIArticle
http://www.sciencedirect.com/science/article/pii/0022283669903702PublisherArticle
Additional Information:© 1969 Elsevier Ltd. Received 31 March 1969. This research was supported in part by a grant GM13554 from the U.S. Public Health. Service.
Funders:
Funding AgencyGrant Number
U.S. Public Health Service (USPHS)GM 13554
Issue or Number:3
Record Number:CaltechAUTHORS:20160802-121819629
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20160802-121819629
Official Citation:John Sedat, Alexander Lyon, Robert L. Sinsheimer, Purification of Escherichia coli pulse-labeled RNA by benzoylated DEAE-cellulose chromatography, Journal of Molecular Biology, Volume 44, Issue 3, 28 September 1969, Pages 415-434, ISSN 0022-2836, http://dx.doi.org/10.1016/0022-2836(69)90370-2. (http://www.sciencedirect.com/science/article/pii/0022283669903702)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:69392
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:03 Aug 2016 16:56
Last Modified:03 Oct 2019 10:21

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