Cloning and Functional Analysis of Zebrafish (Danio rerio) Phosphatidylserine Receptor Promoter

• Main Authors: Chih-Yu Pai, 白植友
• Other Authors: Jiann-Ruey Hong
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/00949687417576626743

碩士 === 國立成功大學 === 生物科技研究所碩博士班 === 95 === The general function of phosphatidylserine receptor that PSR can recognize the apoptotic bodies for clearance of corpse cells, but they temporally and spatially expressive pattern is still unclear. In this thesis, we cloned and constructed the PSR promoter that fused with green fluoresce protein for directly monitoring the temporal and spatial profile of PSR expression during zebrafish embryogenesis. We demonstrated that the 4kb of upstream sequence of PSR gene, which contain promoter region that can drive report gene EGFP expression in early zebrafish embryos. In the PSRP-4K-EGFP transgenic zebrafish fish line (F1), EGFP was expressed from oocyte to hatch out stages, especially, apparently expressed in somites, brain, eyes, heart, hatching gland and open of urinary duct. Interestingly, we first found that the PSR gene can express in skeleton (at the 14 dpf larvae) and testis or ovary (at adult stages). Furthermore, we constructed the different fragments of PSR promoter for the promoter activity assay. In the luciferase reporter assay, we found that a minimal requirement for PSR promoter activity is located in upstream 300 bps of start codon, and in the 3 kbps of upstream sequence that contain repetitive sequence may exist the enhancer elements. In this study, we discovered the PSR is not only important in embryogenesis, but also involved in skeletogenesis and gonadogenesis in late development stages.