PMID: 3754589May 1, 1986Paper

DNA requirements at the bacteriophage G4 origin of complementary-strand DNA synthesis

Journal of Virology
P F LambertW S Reznikoff

Abstract

An in vivo assay was used to define the DNA requirements at the bacteriophage G4 origin of complementary-strand DNA synthesis (G4 origin). This assay made use of an origin-cloning vector, mRZ1000, a defective M13 recombinant phage deleted for its natural origin of complementary-strand DNA synthesis. The minimal DNA sequence of the G4 genome sufficient for the restoration of normal M13 growth parameters was determined to be 139 bases long, located between positions 3868 and 4007. This G4-M13 construct was also found to give rise to proper initiation of complementary-strand synthesis in vitro. The cloned DNA sequence contains all the regions of potential secondary structure which have been implicated in primase-dependent replication initiation as well as additional sequence information. To address the role of one region which potentially forms a DNA secondary structure, the DNA sequence internal to the G4 origin was altered by site-directed mutagenesis. A 3-base insertion at the AvaII site as well as a 17-base deletion between the AvaI and AvaII sites both resulted in loss of origin function. The 17-base deletion was also generated within the G4 genome and found to dramatically reduce the infectious growth rate of the resulting p...Continue Reading

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Citations

Apr 1, 1993·Molecular & General Genetics : MGG·F GiglianiP A Battaglia
Oct 17, 2015·Annual Review of Genetics·Christopher M Johnson, Alan D Grossman
Dec 2, 2010·Microbiology and Molecular Biology Reviews : MMBR·David BikardDidier Mazel

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