The removal of circulating androgens by castration has a marked effect on the physical size, protein, RNA and DNA contents of the rat ventral prostate. This androgen ablation also produces major changes in overall gene expression. Several castration induced mRNAs have previously been identified of which the major sequence is TRPM-2. The cDNA for the rat TRPM-2 has been previously cloned and its protein product characterized. This mRNA sequence is 1,640 bases in length. It codes for a protein of 70-80 kDa, that appears to play an important role in active cell death by blocking complement mediated cell lysis. TRPM-2 is present in a wide variety of species including rat and human. I have isolated genomic clones from two rat and human genomic libraries made from partial Mbo I digests in the EMBL3 vector. I have screened those libraries with the full length cDNA sequence corresponding to the rat TRPM-2, and have isolated overlapping clones which span the TRPM-2 locus in both species. The TRPM-2 gene has been characterized in both species by complete sequence analysis. The gene covers 13.750 bp in the rat and 16.570 bp in human, each having 9 exons. 5,700 bp of upstream sequences have also been sequenced for the rat gene and 1,300 bp for the human. The sequence similarity between the respective coding and 5$\sp\prime$ regions of the gene in both species is nearly 79%. There is also a striking exon/intron structural similarity between the two species. The rat TRPM-2 gene was also shown to be expressed in a wide variety of tissues at very different levels. Finally, the previously published sequence of SGP-2 cDNA is very similar to the TRPM-2 cDNA sequence except for their respective 5$\sp\prime$ ends. PCR analysis has clearly demonstrated that the SGP-2 leader is a cloning artifact.
| Identifer | oai:union.ndltd.org:uottawa.ca/oai:ruor.uottawa.ca:10393/6870 |
| Date | January 1993 |
| Creators | Pineault, Jean M. |
| Contributors | Tenniswood, M., |
| Publisher | University of Ottawa (Canada) |
| Source Sets | Université d’Ottawa |
| Detected Language | English |
| Type | Thesis |
| Format | 149 p. |
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