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To validate this procedure, we calculated conservation of constitutively spliced internal exons at varying levels of ESTs coverage (Table 1).
The phyloP program, using the original alignments and the neutral model from phyloFit, calculated conservation or acceleration P-values for the likelihood ratio test (LRT).
Similarly, we also calculated absolute difference value of structural profiles at each position between every possible pairs of TFBSs, and then calculated conservation rate of DNA structure at each position of TFBS.
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It also implies that the current conservation studies may have missed many conserved regions by calculating conservation scores based on genome alignments.
This calculates conservation of aligned column over background probability distribution.
Base-by-base conservation scores for human bases were downloaded from UCSC Conservation Track [47] which used phastCons [48] to calculate conservation scores across 28 mammalian species.
This program calculates conservation scores for sites in a protein, based on a sequence alignment and phylogenetic trees, through an empirical Bayesian approach.
While ConSurf calculates conservation scores on the basis of multiple sequence alignment typically of hundreds of homologous protein sequences, ProBiS usually uses tens of locally similar structures.
To calculate conservation profiles along a protein sequence, BLASTP [ 64] was used to detect homologous protein sequences from the NCBI protein database.
This may reflect the fact that the two Schizosaccharomyces clade members used to calculate conservation are more distantly related to Sz. pombe than the Saccharomyces sensu stricto clade members are to S. cerevisiae.
The strategy based on the analysis of breed groups produced an overall F st value of 0.048, which was used in the formula of Ollivier and Foulley [ 37] (PCFst = PC Weitz * 0.048 + PCHe*0.952) to calculate conservation values.
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