In vitro complementation as an assay for new proteins required for bacteriophage T4 DNA replication: purification of the complex specified by T4 genes 44 and 62

Proc Natl Acad Sci U S A. 1972 Sep;69(9):2717-21. doi: 10.1073/pnas.69.9.2717.

Abstract

We have developed an in vitro complementation assay for six T4 bacteriophage gene products believed to be components of the T4 DNA replication apparatus. This assay is based upon the fact that DNA synthesis in an infected cell lysate that lacks a given gene product is specifically stimulated by addition of the missing product. By the use of such an assay, two proteins that appear to be the products of T4 genes 44 and 62 have been purified to electrophoretic homogeneity as a single complex of the two polypeptide chains.

MeSH terms

  • Adenosine Triphosphate
  • Bacterial Proteins / pharmacology
  • Carbon Isotopes
  • Chromatography
  • Chromatography, DEAE-Cellulose
  • Coliphages* / drug effects
  • DNA, Viral / biosynthesis
  • Electrophoresis
  • Escherichia coli / drug effects
  • Escherichia coli / metabolism
  • Genes*
  • Genetics, Microbial
  • Hydroxyapatites
  • Isoelectric Focusing
  • Leucine / metabolism
  • Macromolecular Substances
  • Methods
  • Molecular Weight
  • Mutation
  • Sodium Dodecyl Sulfate
  • Tritium
  • Viral Proteins* / biosynthesis
  • Viral Proteins* / isolation & purification

Substances

  • Bacterial Proteins
  • Carbon Isotopes
  • DNA, Viral
  • Hydroxyapatites
  • Macromolecular Substances
  • Viral Proteins
  • Tritium
  • Sodium Dodecyl Sulfate
  • Adenosine Triphosphate
  • Leucine