Temperature affects the sexual differentiation and the development of brain of tilapia, Oreochromis mossambicus. Expressed sequence tags ( EST ) derived from the developing tilapia brain had been cloned in our laboratory. In the present study, we focus on the promoter of cold-induced protein. The promoter sequence of cold-induced protein from the transcription start site ( TSS ) were cloned by 5¡¦ rapid amplification of cDNA end ( 5¡¦-RACE ) and Genome Walker DNA Walking. The bioinformatic analysis was performed on the fragment for predicting the transcription factor binding site. We used the digestion method of restriction enzymes and an electrophoresis mobility shift assay to find transcription factor binding site. The results indicated that there is a putative POU3F2 binding site in the fragment form -157 bp to -149 bp. The luciferase activity assay was performed on this site and results indicated that wild type showed the enhanced promoter activity. However, site-direct mutagenesis of this site did not result in the reduction of the promoter activity.
| Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0902108-165326 |
| Date | 02 September 2008 |
| Creators | Lin, Hou-chi |
| Contributors | Juei-Tang Cheng, Li-Hsueh Wang, Ching-Lin Tsai |
| Publisher | NSYSU |
| Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
| Language | Cholon |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0902108-165326 |
| Rights | not_available, Copyright information available at source archive |
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