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In most of these methods, prior to prediction, alignment positions showing a gap in the top sequence are deleted, consequently leading to shrinking of the alignment and loss of position-specific information.
This is presumably due to erroneous prediction: alignment with other FIKKs clearly shows these sequences represent two parts of a single member of the FIKK family rather than two separate genes.
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For secondary structure prediction alignments were submitted to Quick2D at the MPI Toolkit WWW-server [32] which returns predictions from four independent algorithms.
The p-value cutoff was set to 0.001 in order to reduce false positives prediction alignments.
To assess amino acid polymorphisms for potential functional significance, proteins were compared using Ensembl ortholog prediction alignments between M. musculus and Homo sapiens, Pan troglyodytes, Gallus gallus, Rattus norvegicus, Canis familiaris, Bos Taurus, Xenopus tropicalis, Takifugu rubripes, and Monodelphis domestica when protein homologies existed.
For each pairwise LCB in the true alignment, we record a TP LCB prediction when the predicted alignment contains at least one correctly aligned nucleotide pair in that LCB.
AK performed secondary structure predictions, alignment calculations and phylogenetic reconstructions under supervision of MWo.
The tBLASTx searches, protein prediction, CLUSTALW2 alignments, Pal2Nal alignments, and Ka/Ks calculations were executed using a custom made python script (available on DRYAD).
Because structure prediction and alignment of RNAs depend on each other, Sankoff's approach solves the alignment and folding problem simultaneously.
Chimera [46] used the prediction and alignment information to create the highlighted pdb images.
If this probability is >0.5 (the closer to 1.0 this value is, the more confident is the prediction), the alignment is classified as "functional RNA", indicating that the sequence is from a potential RNA element in ncRNAs or a cis-regulatory element of mRNAs.
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