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Perhaps because of late replication, telomere length is particularly sensitive to mutations in conventional replication proteins.
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This coupled with the loss of late-replication patterns by BrdU IF in ORCA-depleted cells made us hypothesize that ORCA could also regulate the replication of late-replicating regions.
This indicates a loss of late-replication-timing control for the plasmid containing the 8-kb THF in cds1Δ cells, similar to the loss of late-replication-timing control for TAS in checkpoint-deficient cells in their chromosomal context.
We also examined the location of the late replication site in Zsm and Zm using BrdU incorporation assay.
Loss of ORCA could be causing either changes in replication timing of late-replicating regions or the complete loss of replication of these regions.
It is important to stress that the notion of early and late replication origin has intrinsic statistical nature because firing times come from an averaged replication profile deriving from a population-based microarray.
More interestingly, the association of CNV and late replication is distinct for singletons and PDGs.
Cells were arrested at 5 or 9 h post BrdU incorporation, indicative of early or late replication timing respectively (32).
The corresponding flow cytometry results show that these times correspond to late S phase, consistent with the observed late replication of the THF in the chromosome.
These similarities suggest that the Rif1 protein may be at least partially responsible for late replication of the THF.
Similarly, in this study we found that late replication of the THF-containing circular plasmid also requires the Cds1 kinase.
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