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. Transcript levels for various enzymes and proteins involved in murine cellular glycosylation and recognition have been quantitated by our qRT-PCR method allowing medium-throughput (>800 genes) transcript analysis. Parallel data was obtained by Dr. Steve Head, director of the Gene Microarray Core for the Consortium for Functional Genomics, who has developed a microarray chip that allows the screening of mRNA expression patterns for """"""""glycan-related"""""""" genes in mouse cells and tissues. Poly(A+) mRNA and total RNA have been prepared from mouse ES cells and differentiated """"""""embroid bodies"""""""", as well as several mouse tissues, and sent to the Gene Microarray Core for analysis of transcript expression with both their version 2 chip and more recently their version 3 chip. We have compared the results with equivalent data from our real-time RT-PCR analysis and determined that our qRT-PCR approach is far superior to the microarray approaches. Ultimately, the transcript levels will be compared with the glycan structures associated with glycoproteins and glycolipids from the equivalent cell populations and all of the information will become part of the bioinformatics module for display and further correlation analysis.
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