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Independent evidence that differences in cellular abundance contribute to expression variation can be seen by evaluating the correlation structure of transcript abundance across all of the 103 CD, 26 C-type lectin, and 11 Toll-like receptor probes that showed expression above background.
The choice can also be based on the evolutionary conservation of either the exon/intron structure of transcript isoforms or the protein isoforms [ 54].
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These results, together with the observations regarding the petunia CHS-A gene [24, 36, 37, this study], suggest that a conserved feature in exon 2 of the CHS gene across plant species, e.g., the secondary structure of transcripts and/or termination of transcription, is a key element involved in the induction of CHS RNA degradation.
Compared with DGE analysis, the RNA-Seq approach is more powerful for unraveling transcriptome complexity, and for identification of genes, structure of transcripts, alternative splicing, non-coding RNAs, and new transcription units.
Furthermore, RNA-seq can detect genes expressed at low levels and refine the structure of transcripts (Wang et al. [2009] Wilhelmm and Landry [2009]).
Unlike promoter activity, which primarily regulates the amount of transcripts produced by a gene, alternative splicing changes the structure of transcripts and their encoded proteins.
In addition, the gene model that hypothesizes the structure of transcripts produced by a gene also affects the analysis.
Many comparative genomics projects have focused on sequencing of the genome or expressed sequenced tags (ESTs), and full-length cDNA sequences are uniquely informative resources for accurately predicting the full structure of transcripts in the genome [ 15].
Moreover, information regarding the genomic context, including the genomic position, transcription 'start' and 'end' and transcription orientation, is provided, as well as a graphical representation of the exon intron structure of transcripts.
RNA-seq can not only determine the absolute gene expression levels with lower variation compared to microarray technology, but can also be used to find new genes and resolve the structure of transcripts [ 1, 2].
Although there are many complications for variant annotation, we identify two major components: 1. Transcript set: a summary of information about genomic features, particularly the structure of transcripts (sequence and locations of exons, introns, UTRs and regulatory regions), used as the basis for determining the likely functional consequence of a variant.
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