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Using EST data, we identified three chimeric genes that have EST sequences covering all the breakpoints and one chimeric gene that has EST sequence covering some breakpoints.
The importance of RNA-Seq in the detection of novel splice junctions is not in these ubiquitous highly expressed genes, which generally have EST coverage, but in the tissue-specific genes with lower transcript abundance.
Additional support for the role of SNPs in TE population-specific exonization is given by the ssSNPTarget database (http://sssnptarget.org/) [45], the SNPs rs2377301 and rs5758111 have EST evidence for exon skipping due the SNP modification.
Many of the 469 candidate STEPs not reported to have EST evidence of alternative splicing may exhibit an effect leading to an upstream transcript start (where an upstream exon is added by activation of an upstream splice site), intron extension (by activation of a weaker splice site in the adjacent exon) or exon skipping.
Analyses of In100-containing sequences annotated with further upstream and downstream exons in 1252 genes revealed 526 genes that have EST evidence of alternative splicing, showing that the presence of In100 elements correlates with the skipping of their immediate upstream exons.
Without filtering, there are 21,989 sequences that have EST alignments, which is only 9.3% more than the number of sequences with filtered EST alignments (20,123 sequences) and which represents only a 6% increase in the percentage of the hypothetical proteome that is supported by experimental EST evidence.
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By correlating available EST data with gene structure, only genes predicted to have both 5' and 3' UTRs are expressed, although some members with only one UTR have ESTs available.
As shown in Figure 4B, a higher percentage of transcripts that are differentially regulated on the array have EST-support in leaves compared to other organs.
In particular, the amoebozoan Hartmannella vermiformis and the dinoflagellate K. micrum both have EST-predicted TKs related to a proteobacteria-planctomycetes-CFB bacterial clade (a relationship that is further supported by an insertion: Figure 4), and that constitute a weakly supported clade.
All four genes featured in Fig. 6 have ESTs indicating their expression [ 38].
In particular, they showed that 63 (96.9%) of the 65 significantly slowly evolving members have ESTs, whereas 89 (93.7%) of the 95 moderately and rapidly evolving members of animals and plant species did not have ESTs.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com