Studies on in vitro DNA synthesis. Purification of the dna G gene product from Escherichia coli.
varphiX174 DNA-dependent dNMP incorporation is temperature-sensitive (ts) in extracts of uninfected E. coli dna A, B, C, D, E, and G ts strains. DNA synthesis can be restored in heat-inactivated extracts of various dna ts mutants by addition of extracts of wild-type or other dna ts mutants. A protein that restores activity to heat-inactivated extracts of dna G ts cells has been extensively purified. This protein has also been purified from dna G ts cells and is thermolabile when compared to the wild-type protein. The purified dna G protein has a molecular weight of about 60,000, is insensitive to N-ethylmaleimide, and binds poorly to DNA. It does not stimulate heat-inactivated crude extracts of dna B, C, D, or E ts cells and lacks detectable RNA and DNA polymerase activities.
D000645:Ammonium Sulfate; D001426:Bacterial Proteins / Q000302:isolation & purification*; D001665:Binding Sites; D002499:Centrifugation, Density Gradient; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004261:DNA Replication; D004269:DNA, Bacterial / Q000096:biosynthesis*; D004229:Dithiothreitol; D004492:Edetic Acid; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D005615:Freezing; D005796:Genes; D005816:Genetic Complementation Test; D005990:Glycerol; D008722:Methods; D009154:Mutation; D013307:Streptomycin; D013431:Sulfates; D013696:Temperature
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D013431:Sulfates; D004492:Edetic Acid; D005990:Glycerol; D000645:Ammonium Sulfate; D004229:Dithiothreitol; D013307:Streptomycin