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A typical sequence processing pipeline includes several steps such as sequence cleaning, alignment to known reference sequences, and/or de novo assembly [2], [10].
Transcripts were identified as PCWDEs based on amino acid alignment to known proteins.
miRNAs are often identified by cloning or by bioinformatic alignment to known miRNAs.
To filter out non-paralogous regions different sequence identity cutoffs in the ClustalW alignment to known Human H/ACA snoRNAs were applied.
GreenGenie2 predictions on Chlamydomonas genome assembly v3 were screened for a minimum coding length of 270 bp and against significant alignment to known transposable elements (see Methods).
This allowed us to identify some models as being artifactual due to misalignment or redundant alignment to known loci, excluding these from further analysis.
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The remaining two cases were relatively weak hits (108 and 103 bits) that were not captured by the alignments to known families.
Putative genes, i.e., sequences exhibiting significant alignment (S' ≥50) to known genes and/or cDNA sequences, constitute approximately 4.1% of the BAC sequence (including the putative LEA gene – see below).
Seventy-two percent of the sequences encoded putatively full-length GH11 polypeptides based on alignment length to known GH11 polypeptides and the presence of in-frame putative start and stop codons.
The trypsin/toxin complex model is based mainly on the structural alignment to the known coordinates of the BPTI/toxin complex (PDB: 1TAW).
These conformations served as starting positions for further docking, each time selecting a peptide with similar or better ICM docking score and a superior binding conformation by backbone alignment to the known GSK3β peptide conformation.
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