SRD5A1 Genetic Variation and Prostate Cancer Epidemiology
Abstract
Seven 3' untranslated region(UTR) single nucleotide polymorphisms (SNPs) and one double mutant from the human steroid 5-alpha reductase I gene were functionally tested indirectly for mRNA stability using a luciferase construct in a human kidney cell line. There was no difference in luciferase signal Using any of the variants, but the 3 `UTR did double luciferase signal suggesting a stabilization effect. It was hypothesized that the secondary structure of the luciferase+SRD5Al 3'-UTR mRNA was so different from the native SRD5Al message, that the 3'-UTR variants should be tested in the context of a native mRNA. A eukaryotic expression vector containing 850bp of the human SRD5Al promoter was seamlessly cloned onto the 5' end of the full length SRD5Al cDNA (containing the full length UTR). Human beta-globin intron 2 was inserted into exon 4 using a native blunt restriction site to ensure proper mRNA processing and export. This vector has been constructed and is being evaluated for expression via northern blotting and RT-PCR. There are no studies evaluating how SNPs in `RNA haplotypes' alter RNA half-life, stability, or protein translatability.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2005
- Accession Number
- ADA441326
Entities
People
- Troy J. Phipps
Organizations
- University of Southern California