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mutants for ctrA
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Mutants for ctrA, chpT and cckA showed almost homogeneous cell morphology and divided by binary fission.
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The Ct values for the four ctrA-negative, sodC-positive CSF extractions averaged 40.4 for ctrA while their sodC Ct values averaged 32.0.
After DNA extraction and real-time PCR testing of all specimens for ctrA of Nm [5], lytA of S. pneumoniae [22], and bexA and/or bcs2 of Hi, the subset of specimens chosen to test the sodC assay (n = 120) were either (1) positive for ctrA (n = 12) or (2) ctrA− lytA− bexA/bcs2− (n = 108).
The remaining 12 CtrA target genes, whose transcript levels remain constant during cell cycle progression in complete media, point to a novel role for CtrA in activating and repressing genes involved in the response to starvation and other stress signals, in addition to those involved in cell cycle progression.
Binding sites for CtrA were found in the promoter of ctrA itself, suggesting an auto-regulatory loop for this gene.
Both GcrA and DnaA were then analyzed as reported in points 2 and 3 for CtrA.
Conservation of dnaA in all Wolbachia strains analyzed also supports an important role for CtrA in regulating genome replication.
An essential phosphorelay, composed of the hybrid histidine kinase CckA and the histidine phosphotransferase ChpT, is responsible for CtrA phosphorylation [ 20, 21].
Four specimens had indeterminate RT-PCR results (two for lytA and two for ctrA), and these results were considered negative in all analyses.
We searched for CtrA binding sites in the promoters of all genes that have either an autoinducer synthesis (pfam00765) or an autoinducer binding (pfam03475) domain.
This indicates that long 5′-UTRs are not unusual in S. meliloti and may even signify complex mechanisms of gene regulation, as is likely the case for ctrA.
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