碩士 / 國立海洋大學 / 水產生物技術研究所 / 90 / Proteins of the Bcl2 family play important roles in animal development; however, the underlying molecular mechanisms of their actions are poorly understood. As part of our undergoing effort to develop zebrafish embryos as a model system for the embryonic study of the Bcl2 family proteins, here we described the cloning and characterization of the third members of the zebrafish Bcl2 protein family, zfBax1. The zfBax1 cDNA is 1938 nucleotides long, encoding a polypeptide of 192 amino acids. The primary sequence of zfBax1 shares 54 % identity to human Baxisoform, and contains all three conserved BH domains of the pro-apoptotic Bcl-2 family proteins. Northern blot analysis detected a single mRNA species with decreasing abundance over time in oocytes and all selected stages of developing embryos. GFP-tagging analysis demonstrated a punctate cytoplasmic distribution pattern, which was overlapping with mitochondria-specific staining for the tagged zfBax1 protein in transfected cells. Over-expression of GFP-zfBax1 protein resulted in rapid and drastic cell death in transfected cells even before the expression of the fusion protein could be detected by epi-fluorescent microscopic examination. Finally, GFP-zfBax1 over-expression led to apoptotic death in embryonic cells as apparently by nuclear condensation and DNA fragmentation, and its killing effect was retarded by simultaneous over-expression of GFP-zfmcl1a protein. Overall, our results suggest that zfBax1 is a pro-apoptotic Bcl-2 family protein, and is likely to play an important role in zebrafish oogenesis and embryogenesis.
| Identifer | oai:union.ndltd.org:TW/090NTOU0613001 |
| Date | January 2002 |
| Creators | Jia-Pey Wang, 王嘉佩 |
| Contributors | Jen-Leih Wu, 吳金洌 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 53 |
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