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博碩士論文 etd-0912107-171547 詳細資訊
Title page for etd-0912107-171547
論文名稱
Title
人類ROGDI蛋白之功能性分析
Functional analysis of human ROGDI protein
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
76
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2007-07-23
繳交日期
Date of Submission
2007-09-12
關鍵字
Keywords
細胞增生
ROGDI
統計
Statistics
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中文摘要
ROGDI是從人類腎臟上皮細胞所選殖出來的一個基因,其位於人類染色體16p13.3上,遺傳編碼區為864 bp,轉譯出蛋白質有287個胺基酸。本實驗室之前的研究指出在許多癌細胞株中ROGDI的表現比正常細胞高。藉由細胞生長曲線得知在HEK293T以及Hep3B細胞中過量表現ROGDI時會促進細胞生長。另一方面,根據間接免疫螢光染色的結果發現部分ROGDI會於細胞核內表現,因此猜測ROGDI可能涉及細胞週期的調控。為了支持這個論點,以流式細胞儀分析細胞週期的變化,顯示降低ROGDI的表現會使細胞週期停留在G1 期的HeLa 細胞明顯變少。此外,不論是以BrdU 偵測DNA合成速率或是以WST-1測試粒線體中酵素活性,過量表現ROGDI的HEK293T細胞之DNA合成速率及細胞代謝活性都比控制組細胞快。綜合上述我們推測ROGDI可能是一個細胞週期的正調控因子,促進細胞週期向前推進。
Abstract
ROGDI was a novel gene isolated from primary human renal epithelial cells. The gene located on chromosome 16p13.3 and the size of coding region is 864 bp which encodes 287 amino acids. Our previous studies revealed that the expression of human ROGDI gene is elevated in several cancer cell lines. Overexpression of ROGDI increases proliferation in the HEK293T and Hep3B human cell lines, as indicated by cell growth curves. On the other hand, according to indirect immunofluorescence, some ROGDI expressed in the nucleus. Therefore, we suggest that it may involve in regulation of cell cycle. In support of this, flow cytometry studies using HeLa cells demonstrate that ROGDI decreases the overall population of cells in G1 phase of the cell cycle. Furthermore, cells overexpressing ROGDI demonstrate an increase in the incorporation of BrdU and in WST-1 activity, respectively, indicating that both DNA synthesis and mitochondrial enzyme activity are enhanced. Taken together, ROGDI appears to be a positive regulator of cell cycle progression.
目次 Table of Contents
中文摘要 1
英文摘要 2
壹、 前言 3
一、 ROGDI 3
二、 細胞週期 4
三、 核醣核酸干擾現象 6
四、 重組人類反轉錄病毒 9
貳、 實驗材料與方法 12
參、 實驗結果 31
肆、 討論 39
參考資料 42
圖表 47
附錄 66
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