Identification of Novel Secreted Molecules of Prostate Cancer
Annual rept. 1 Jun 98-31 May 99
CEDARS-SINAI MEDICAL CENTER LOS ANGELESCA
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We have developed an approach to isolate new tumor markers of prostate cancer. In order to identify genes encoding secreted protein, we have modified the signal sequence trap technique, using the SUC 2 gene encoding a secreted protein of yeast. Only the yeast which have been transformed with cDNAs containing secretory signal sequences can grow into colonies on the plates which contain sucrose as the only source of sugar. Early difficulties identifying positive clones with our initial vector system were problematic we obtained a new vector system to reduce the possibility of enzymatically inactive fusion invertase and enhance the sensitivity of this signal trap system which allowed us to obtain positive clones. We successfully constructed and subtracted a prostate specific cDNA library. We will continue to pursue the selection of positive clones, make RNA dot blots from various tissues to establish tissue specificity of the cDNA and perform homology searches to determine if we have found a novel gene. Unique genes will be cloned into a GST vector and fusion proteins will be made and used to make antibody against the protein. Serum sample assays will be developed from antibodies that identify prostate specific genes. Additionally, structurefunction analysis and sense and antisense expression vectors will be employed to elucidate further the function of these genes.
- Medicine and Medical Research