請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75320
標題: | 鯉魚減數分裂染色體濃縮相關基因Nek2、Ran之選殖 Molecular cloning of chromosome condensation related genes, Nek2 and Ran, in the carp oocytes |
作者: | Hsin-Yi Chen 陳心怡 |
出版年 : | 2002 |
學位: | 碩士 |
摘要: | 細胞進行有絲分裂時,染色體必須先複製、濃縮,再經由分離染色體的母機—紡錘體,將之分到兩個子細胞中。進行減數分裂的細胞也必須歷經以上過程,然而由於染色體組態及紡錘體行為的差異,減數分裂必須有所修飾或特化。研究顯示,Nek2和Ran與染色體濃縮皆有直接的關聯,Ran更為紡錘體形成機制的一員,而減數分裂特有蛋白Mos可能透過作用到相關元件,影響減數分裂染色體組態及紡錘體的形成、變動與維持。 細胞分裂時染色體、紡錘體狀態的控制元件彼此之間勢必需要直接的相互作用或間接的溝通,為了探究減數分裂細胞染色體組態變化及紡錘體行為特殊風貌的分子機制,瞭解相關的元件為第一步,因此本實驗自鯉魚的卵巢選殖了Nek2、Ran基因,於細菌表現Nek2重組蛋白,並製作了anti-Nek2抗血清,加上先前實驗室已選殖、表現的Mos基因,希望以此做為建立in vitro研究系統的基礎。由於Mos表現蛋白出現自我聚集的現象,為了探討此現象是否為內生性性狀,本實驗也於細菌表現了失去激?活性及C端truncated突變種Mos蛋白,希望能夠藉此瞭解Mos的性質並獲得有活性的表現蛋白,建立in vitro assay系統,釐清Nek2、Ran及Mos蛋白在減數分裂的相互關係。 Chromosome condensation and spindle formation are the principle phenomena of mitosis. Compared with mitosis, meiosis has the particular characters in chromosome configuration and spindle behavior. However, chromosome condensation and spindle formation are still necessary meiotic process. Because of the different between mitosis and meiosis, the meiotic machine must be modified. Nek2 and Ran had be shown to involve in chromosome condensation. Furthermore, Ran participates in spindle formation. On the other hand, Mos knockout data implied that the vertebrate meiosis protein is related to chromosome status and microtubule dynamics. For the sake of completing the separation, cell chromosome condensation components have to coordinate, via either direct interaction or indirect communication. To investigate the characteristic of meiosis molecular mechanism, the cDNA of Nek2 and Ran genes had been cloned from the carp ovary. The carp Nek2 recombinant protein expressed in bacteria was used to produce Nek2 antiserum. Combining with the previous cloned Mos, it will be the basis of in vitro assay system. When expressing Mos recombinant protein, we observed that the recombinant protein tended to aggregation. To inquiry if this is Mos intrinsic characteristic, kinase-dead and C-terminal truncated mos mutant were also expressed in bacteria. In the expect of grasping Mos property, obtaining active expression protein and establishing a in vitro assay system to deliberate the connection between Mos, Nek2 and Ran in the future. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75320 |
全文授權: | 未授權 |
顯示於系所單位: | 動物學研究所 |
文件中的檔案:
沒有與此文件相關的檔案。
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。