Fidelity of DNA replication catalysed in vitro on a natural DNA template by the T4 bacteriophage multi-enzyme complex.
More than 50 copies of a phi X174 DNA template can be made in 60 min in an in vitro DNA replication system consisting of seven purfied replication proteins isolated from T4 bacteriophage-infected cells. By transfecting with the DNA products and assaying for the reversion of specific amber mutants, the high degree of base-pairing fidelity in this system is revealed; the in vitro system is also shown to respond to the mutagenic effect of Mn2+ and to display strong base-pair context effects on fidelity, as expected from in vivo studies.
D010584:Bacteriophage phi X 174 / Q000235:genetics; D001483:Base Sequence; D004265:DNA Helicases / Q000378:metabolism; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000378:metabolism; D005092:Exonucleases / Q000378:metabolism; D008345:Manganese / Q000494:pharmacology; D009097:Multienzyme Complexes / Q000378:metabolism*; D013604:T-Phages / Q000235:genetics*; D013698:Templates, Genetic
D004279:DNA, Viral; D003854:Deoxyribonucleotides; D009097:Multienzyme Complexes; D008345:Manganese; D004259:DNA-Directed DNA Polymerase; D005092:Exonucleases; D004265:DNA Helicases