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標題: | 鯉魚卵巢p18之純化、定性與組織分佈 Purification、Characterization and Immunocytochemistry of p18 from Carp Ovary |
作者: | 張志聖 |
出版年 : | 2001 |
學位: | 碩士 |
摘要: | 卵膜是包圍在卵細胞的外殼,對於魚類而言,有著重要的生理意義,它能保護魚卵不遭受環境中各種物理性或化學性傷害,也能避免魚卵受到細菌等病源的感染。在表層反應發生後,儲存於表層粒中的蛋白或酵素會釋放到細胞外的卵膜上,並進行重組成為新的卵膜,堅硬的新卵膜更可以抵禦環境對卵細胞的傷害。本實驗室致力於鯉魚卵膜組成的研究,過去已發現多種組成分子,包括p25、p30、ZP2、ZP3、cystaitin、cathepsin-like substance、fibroin-like substance、FEO-l,並對其性質加以探討,但卵膜上還有許多尚未被發現之組成分子。 本論文在鯉魚的卵膜萃取液中,發現一個分子量為18kDa的蛋白,暫命名為P18,並利用各種純化方法分離P18,分析其化學性狀,並誘導了抗體。在西方轉印法及免疫組織化學法的實驗中,均發現表層反應前的卵膜即有P18的存在,隨著表層反應的發生,P18在卵膜上的量也逐漸的的增強,顯示P18不但是卵膜的成員,也參予新卵膜的組成。 除此之外,我們也發現P18在卵膜上並非以單體(monomer)分子形式存在,而是與其他分子交互連結,然而究竟有哪些分子與P18連結,則需更進一步的探討。 Chorion is an envelope surrounding oocyte. It is a critical structure in most fish species that can protect egg from physical and chemical damage, and microbial infection. When cortical reaction takes place, the cortical granular components discharge into chorion. During cortical reaction, the proteins and enzymes of cortical granules are recruited to the chorion. They will be reconstructed with the pre-existing components of chorion to form fertilization envelope (FE). The harder FE can shelter embryos from environmental damage. Our laboratory has long been devoted to study in the carp chorion's study. Several components have been identified, including p25、p30、ZP2、 ZP3、cystatin、cathepsin-like substance、 fibroin-like substance、 FEO-1. However, there are still a lot of unidentified components on chorion. In this study , we purified and characterized a new component of chorion, p 18 , from carp eggs, The result of immumocytochemical staining reveals that p18 preexists on carp chorion before cortical reaction , but the amount of p18 on FE is higher than that of chorion. These results indicate that p18 is not only the component of chorion, but also participates in the formation of FE. Moreover, p18 exists as multiforms on chorion by cross-linking with other molecules. However, which molecules are cross-linked with p18 remain to be elucidated. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75245 |
全文授權: | 未授權 |
顯示於系所單位: | 生化科學研究所 |
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