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The Replication of Bacteriophage MS2: VIII. Fractionation of the “Replicative Intermediate” using a BNC Column

Kelly, Regis B. and Sinsheimer, Robert L. (1967) The Replication of Bacteriophage MS2: VIII. Fractionation of the “Replicative Intermediate” using a BNC Column. Journal of Molecular Biology, 29 (2). pp. 229-236. ISSN 0022-2836. http://resolver.caltech.edu/CaltechAUTHORS:20160531-143407861

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Abstract

The template on which single strands of MS2 RNA are synthesized, often called the replicative intermediate, is a partially double-stranded molecule. This unusual secondary structure permits the purification of the replicative intermediate on columns of benzoylated, naphthoylated DEAE cellulose (BNC), which have been shown to fractionate nucleic acids with respect to the extent of their double-stranded character. The replicative intermediate is eluted from such a column after DNA and before single-stranded MS2 RNA, but shows heterogeneity. The fractions of replicative intermediate which are eluted at higher sodium chloride concentrations are more sensitive to RNase and have a buoyant density closer to that of single-stranded RNA. The results suggest that the replicative intermediate molecule consists of a double-stranded “core” to which is attached one singlestrand tail of variable length and that these molecules are fractionated on the BNC column with respect to their tail length. All the fractions of replicative intermediate are infective upon denaturation.


Item Type:Article
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URLURL TypeDescription
http://dx.doi.org/10.1016/0022-2836(67)90213-6DOIArticle
http://www.sciencedirect.com/science/article/pii/0022283667902136PublisherArticle
Additional Information:© 1967 Elsevier Ltd. Received 28 April 1967. We gratefully acknowledge the advice and criticism of Mr John W. Sedat. This work was supported in part by research grant GM 13554 from the U.S. Public Health Service.
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U.S. Public Health Service (USPHS)GM 13554
Record Number:CaltechAUTHORS:20160531-143407861
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20160531-143407861
Official Citation:Regis B. Kelly, Robert L. Sinsheimer, The replication of bacteriophage MS2, Journal of Molecular Biology, Volume 29, Issue 2, 1967, Pages 229-236, ISSN 0022-2836, http://dx.doi.org/10.1016/0022-2836(67)90213-6. (http://www.sciencedirect.com/science/article/pii/0022283667902136)
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ID Code:67498
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:31 May 2016 23:13
Last Modified:31 May 2016 23:13

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