TRANSCRIPTOME 2002: From Functional Genomics to Systems
From Transcriptome to Proteome: Genome Scale Collection and Expression of Low Abundant Human Full-Length Transcripts
Yijun Ruan, Chia-Lin Wei, Stephan Dillard-Telm, Erika Peterson, Gina Truesdell, Helen Chan, Gary Wolfe, and Dave Mannion, Large Scale Biology Corporation, Vacaville, CA
A fundamental aspect of understanding human genome is to clone, sequence, and analyze all of full-length transcripts including their alternative splicing variants. A complete collection of full-length cDNA clones and their sequences will not only refine the global view of a genome and detail the structures of genes, but also fuel the whole range of proteomics through high throughput recombinant protein expressions. Recently we developed an efficient process to select low abundant cDNA clones based on their abundance level in transcriptome for sequencing analyses. With this approach we have achieved the highest discovery efficiency of full-length transcript cloning and sequencing. So far we cloned and characterized tens of thousands unique human full-length transcripts from hundreds of human tissues, cell lines, and biopsy samples in expression ready vectors, and identified large number of novel transcripts that have not been described, as well as many new alternative slicing variants of known genes. We are in process of generating thousands of human recombinant proteins using GENEWAREŽ (a plant viral expression system). These high quality collections of low abundant human full-length genes and recombinant proteins will be invaluable resources for gene function and proteomics studies. Detailed methodology and full-length transcripts content will be discussed.
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