Cellular localisation of the clamp protein during DNA replication.
The beta subunit of Escherichia coli DNA polymerase III holoenzyme was fused to the green fluorescent protein GFP. The gene fusion under the control of the heterologous lac promoter was used to replace the wild-type allele in the chromosome. The formation of GFP-beta fluorescent foci in GFP-beta expressing cells required DNA replication and their number per cell was dependent on cell growth. Examination of GFP-beta foci in a synchronous round of replication suggested that DNA replication was accompanied by the recruitment of GFP-beta foci near the midcell, followed by the rapid migration of the foci in opposite directions to the 1/4 and 3/4 positions during DNA replication.
Kongsuwan
Kritaya
K
CSIRO Division of Livestock Industries, 120 Meiers Road, 4068, Indooroopilly, Qld, Australia. kritaya.kongsuwan@csiro.au
Dalrymple
Brian P
BP
Wijffels
Gene
G
Jennings
Phillip A
PA
D002876:Chromosomes, Bacterial; D004258:DNA Polymerase III / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication / Q000502:physiology*; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D049452:Green Fluorescent Proteins; D015151:Immunoblotting; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D016296:Mutagenesis / Q000502:physiology; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D004258:DNA Polymerase III