請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76060
標題: | 食道上皮細胞分化核蛋白之分子選殖及特性之研究 Molecular Cloning and Characterization of A Bovine Esophageal Epithelial Nuclear Protein Involved in Epithelial Cell Differentiation |
作者: | 劉珍 |
出版年 : | 1994 |
學位: | 碩士 |
摘要: | 為研究分化相關的細胞核蛋白質,本實驗室曾以牛食道上皮細胞核萃取物,製備了一系列之單株抗體,並篩選出一株可以專一地認得牛食道上皮末期分化細胞緻密核的單株抗體W2。利用免疫沈澱法分離W2所認得的抗原,得到了-33 kDa的蛋白質複合體。此複合體又可以還原分離為-11kDa及-22kDa的蛋白質,分別稱為牛食道上皮蛋白(Bovine Esophageal Epithelium),BEE11及BEE22。在本論文中,我選殖出BEE11完整之genomic DNA序列,並利用基因資料庫尋找相似的基因進行分析與比對,發現BEE11與一人類白血球分化調節基因,MRP(migration inhibitory factor-related protein)8,在序列比較上有極大的相似性。此外,我利用E. coli expression vector製備了BEE11之重組蛋白,及產生具有高度專一性之多源抗體。以Northern及Western Blot分析BEE11在牛各組織中的表現,顯示BEE11主要分佈在牛食道與牛舌等上皮細胞組織中。 Some nuclear proteins that exhibit unique expression patterns may play important roles in the differentiation program. In order to study the cell differentitaion mechanisms, a series of monoclonal antibodies that against nuclear extracts isolated from bovine esophageal epithelium were raised by our laboratory. One of these monoclonal antibodies, W2, stained specifically at the condensed nuclei of the superficial epithelium, but not the cells at the basal stratum of bovine esophagus. This specific staining pattern suggested that W2 antigen may be involved in epithelial cell differentiation. Immunoisolation of W2 target antigen from the nuclear extracts of superficial cells yielded a 33kDa protein complex named as bovine esophageal epithelium (BEE)33. Upon DTT reduction, BEE33 dissociated into two polypeptides, BEE11 and BEE22. In this study, I have isolated the BEE11 gene from a bovine genomic library and have characterized its genomic structure. My results showed that BEE11 gene shared high degree of similarity with human MRP (migration inhibitory factor-related protein) 8 gene. A polyclonal antibody that specifically against BEE11 protein was also raised. Northern and Western blot analyses were used to investigate the distribution of BEE11 in various bovine tissues at both mRNA and protein levels. My results show that BEE11 appears to be mainly expressed at bovine esophagus and tongue tissues. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76060 |
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。