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Overproduction and Purification of the Gene 2 Product Involved in the Initiation of Phage Φ29 Replication

  • Luis Blanco
  • Juan A. García
  • José M. Lázaro
  • Margarita Salas
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 179)

Abstract

The Bacillus subtilis phage Φ29 has a linear, double-stranded DNA with the protein product of gene 3, p3, covalently linked to the two 5’ ends (1). Φ29 replication starts at either DNA end by a novel mechanism by which the terminal protein p3 primes replication by reaction with dATP and formation of a protein p3-dAMP covalent complex that provides the 3’0H group needed for elongation by the DNA polymerase. The initiation reaction, i.e. the formation of the protein p3-dAMP covalent complex, requires Φ29 DNA-protein p3 as a template, free protein p3 and the gene 2 product (2, 3). We present here the cloning of gene 2 and the overproduction and partial purification of protein p2. A DNA polymerase activity has been found associated with protein p2.

Keywords

Ammonium Sulfate Partial Purification Initiation Complex Initiation Reaction Plasmid pBR322 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 1984

Authors and Affiliations

  • Luis Blanco
    • 1
  • Juan A. García
    • 1
  • José M. Lázaro
    • 1
  • Margarita Salas
    • 1
  1. 1.Centro de Biología Molecular (SDIC-UAM)Universidad Autónoma Canto BlancoMadrid - 34Spain

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