Fukuda, Akio and Sinsheimer, Robert L. (1976) Process of infection with bacteriophage phi X 174 XXXVIII. Replication of phi chi 174 replicative form in vivo. Journal of Virology, 17 (3). pp. 776-787. ISSN 0022-538X http://resolver.caltech.edu/CaltechAUTHORS:FUKjvir76a
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The replication of bacteriophage phi X 174 replicative-form DNA has been studied by structural analysis of pulse-labeled replicative-intermediate molecules. Such intermediates were identified by pulse-labeling with [13H]thymidine and separated into four major fractions (A, B, C, and D) in a propidium diiodide-cesium chloride buoyand density gradient. Sedimentation analysis of each of these fractions suggests the following features of phi X replicative-form DNA replication in vivo. (i) At the end of one cycle of replication, one daughter replicative form (RFII) contains a nascent plus (+) strand of the unit viral length, and the other daughter RFII contains small fragments of nascent minus (-) strand. (ii) Asymmetry is also associated with production of the first supercoiled RFI after addition of pulse label in that only the minus strand becomes radioactive. (iii) A supercoiled DNA (RFI') seems to occur in vivo. This DNA is observed at a position of greater density in a propidium diiodide-cesium chloride buoyant density gradient than normal RFI. (iv) A novel DNA component is observed, at a density greater than RFI, which releases, in alkali, a plus strand longer (1.5 to 1.7 times) than the unit viral length. These results are discussed in terms of the possible sequence of events in phi X 174 replicative-form replication in vivo.
|Additional Information:||Copyright © 1976 by the American Society for Microbiology. Received for publication 2 September 1975 This work was supported by Public Health Service grant GM-13554 from the National Institute of General Medical Sciences.|
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|Deposited On:||16 May 2006|
|Last Modified:||26 Dec 2012 08:52|
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