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Therefore, we can exclude an influence of divergence and number of identified tRNAs on estimates of constraints in our data.
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No qualitative change in the alignments or resulting estimates of constraint was observed.
These standard estimates of constraint and adaptation become more stringent under conditions of genetic linkage.
Weighted estimates of constraint were obtained by a method similar to the approach of Halligan and Keightley (2006).
Based on three-species alignments of chicken, turkey, and zebra finch, lineage-specific estimates of constraint were 43%, 29%, and 24%, respectively.
This selection was applied to exclude the possibility of differences in rate and pattern of nucleotide substitution between regions with and without intronic AR affecting the estimates of constraint.
The MSHAKE algorithm, which is applied by modifying the leapfrog algorithm to include forces of constraint, computes an initial estimate of constraint forces, then iteratively corrects the constraint forces required to maintain the fixed distances.
Therefore, P provides a conservative estimate of constraint.
Using a procedure similar to Halligan and Keightley (2006) (for details, see Material and Methods), we obtained a weighted estimate of constraint of 31.5% (±2.0%) for the 4-fold degenerate sites.
These caveats suggest that the lower estimate of constraint at 4-fold degenerate sites in the zebra finch than in the chicken and turkey lineages should not be taken too far.
The present codon-based model with the ML estimates of selective constraints and with adjustable mutation rates of nucleotide would be useful as a simple substitution model in ML and Bayesian inferences of molecular phylogenetic trees, and enables us to obtain biologically meaningful information at both nucleotide and amino acid levels from codon and protein sequences.
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