This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our group is part of an effort to describe the complete transcriptome and eventually proteome for individual cell types during development in the free-living nematode C. elegans. At this point we and others have made great strides in describing the transcriptome using a number of different techniques including microchips, SAGE and promoter::GFP fusions (http://elegans.bcgsc.bc.ca/). We would now like to set our sights on developing a complete proteome for specific cells during development. We think that a study of the proteome in C. elegans, an organism with a sequenced genome and the extensive background information we have on the transcriptome, is a logical extension of our present work. The two approaches we feel will reinforce one another. In one case the identification of a protein in a tissue where we have identified a transcript will affirm the expression of the gene in that tissue. On the other hand our identification of thousands of transcripts within a tissue gives a benchmark from which to measure the success of protein analysis and the concept of proteomics. What we propose is to start with the whole nematode embryo of 550 cells (representing many tissue types) and a known profile of some 7,000 expressed genes and then move to successively more challenging to prepare tissue samples. After we do the whole embryo we will start with the body wall muscle as this is easily accessible tissue (81 of the 550 embryonic cells) and we can obtain it in large quantities. After gaining experience with this tissue we will move to the more challenging neural tissues.The proteome will be compared to the transcriptome, including identified splice variants, to determine the relative amount and type of post-transcriptional regulation occurring in each cell type. We think it is possible that this collaboration could lead to the fullest description of the proteome in any metazoan.
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