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The Process of Infection with Bacteriophage φX174: XIX. Isolation and Characterization of a Chloramphenicol-resistant Protein from φX-infected Cells

Levine, Arnold J. and Sinsheimer, Robert L. (1968) The Process of Infection with Bacteriophage φX174: XIX. Isolation and Characterization of a Chloramphenicol-resistant Protein from φX-infected Cells. Journal of Molecular Biology, 32 (3). pp. 567-578. ISSN 0022-2836. https://resolver.caltech.edu/CaltechAUTHORS:20160601-093438429

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Abstract

A chloramphenicol-resistant, viral-directed protein, associated with the synthesis of progeny RF‡ in cells infected with bacteriophage φX174 has been isolated. Uninfected and φX174-infected Escherichia coli C were labeled, in the presence of 30 μg/ml, of CM, with [^(14)C]leucine and [^3H]leucine respectively. The labeled cells were combined, lysed and the lysates fractionated by CaHPO_4 chromatography. A radioactive component (or components) preferentially present in the infected cell extracts was found. This protein was not synthesized in the presence of 100 μg/ml. of CM nor was it formed in nonpermissive cells infected with φX amber mutants of cistron VI; progeny RF is not synthesized under either of these conditions. The CM-resistant φX protein was further separated from contaminant E. coli CM-resistant proteins (14C-labeled) by gel fiitration on Sephadex G100. The elution pattern observed with Sephadex G100 was dependent on the pH and ionic strength. Lysis of the cells with lysozyme, EDTA and Brij, a non-ionic detergent, followed by chromatography on CaHPO_4, yielded more homogeneous preparations than lysis with lysozyme, EDTA and freeze-thawing.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/0022-2836(68)90343-4DOIArticle
http://www.sciencedirect.com/science/article/pii/0022283668903434PublisherArticle
Additional Information:© 1968 Elsevier Ltd. Received 14 September 1967, Revised 20 November 1967. This research was supported in part by grant GM 13554 from the U.S. Public Health Service. One of us (A.J.L.) has been supported by a post-doctoral fellowship from the U.S. Public Health Service.
Funders:
Funding AgencyGrant Number
U.S. Public Health Service (USPHS)GM 13554
NIH Postdoctoral FellowshipUNSPECIFIED
Issue or Number:3
Record Number:CaltechAUTHORS:20160601-093438429
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20160601-093438429
Official Citation:Arnold J. Levine, Robert L. Sinsheimer, The process of infection with bacteriophage φX174, Journal of Molecular Biology, Volume 32, Issue 3, 1968, Pages 567-578, ISSN 0022-2836, http://dx.doi.org/10.1016/0022-2836(68)90343-4. (http://www.sciencedirect.com/science/article/pii/0022283668903434)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:67527
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:01 Jun 2016 20:45
Last Modified:03 Oct 2019 10:06

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