出版年:

研究生:

蘇文正

研究生(英文姓名):

Wen-Jenq Su

論文名稱:

分離並鑑定小鼠前胸腺激素啟動子的特性

英文論文名稱:

Identification and characterization of the promoter region of murine prothymosin α

指導教授:

蕭 璦 莉 博士

指導教授(英文姓名):

Ai-Li-Shiau

學位類別:

碩士

校院名稱:

國立成功大學　

系所名稱:

微生物暨免疫學研究所

學號:

S46851128

學年度:

86

語文別:

中文

論文頁數:

65

關鍵詞:

前胸腺激素 ; 綠色螢光蛋白 ; 轉錄因子 ; 細胞週期 ; 啟動子

英文關鍵詞:

prothymesin a ; EGFP ; transcriptional factor ;
cell cycle ; promoter

被引用次數:

0

[ 摘要 ]

前胸腺激素(prothymesin a, Pro T a)為一酸性核蛋白，亦為胸腺激素的一員，扮演著免疫調節者(immunomodulator)及細胞增生(cell proliferation)的角色。在許多的細胞上，都有大量的表現，且可經由C-myc活化。但到目前為止，其生理上的真正功能仍不明確。由其在演化上的完整性(evolutionary conservation) ，及有廣泛的組織分布這二點看來，它在細胞生長的整個過程中，扮演了相當重要的角色。為了研究它的調控機制，以方便未來組織特異性(tissue-specific)的研究，利用綠色螢光蛋白作為報告基因，建構一系列質體，之後再送入NIH 3T3細胞中，測得綠色螢光蛋白表現的量來判讀其啟動子(promoter)的活性。經由G418藥物篩選出穩定表現綠色螢光的細胞株，之後再利用藥物處理，並利用流體細胞分析儀分析，便可得知前胸腺激素啟動子的活性，主要受到轉錄因子的調控，而非與細胞週期有絕對的關係。

[ 英文摘要 ]

Prothymosin a (ProT a) is an acidic nuclear protein that is a member of thymic hormones; it can act as an immunomodulator and has an intracellular function associated with cell proliferation. It is expressed at high levels in a wide variety of cell types and may function in a c-myc associated pathway. Although the exact function of ProTa remains unknown, its highly evolutionary conservation and its wide tissue distribution suggest that it may play an essential role in the cell. In order to study the regulatory mechanisms of ProTa expression, we isolated a series of murine DNA fragments upstream the coding sequence of the murine ProTa gene. These fragments were linked to a reporter gene, the enhanced green fluorescent protein (EGFP), to construct a series of reporter plasmids which were used to transfect into NIH 3T3 cells. The promoter activity of ProTa can be identified by analyzing the expression of EGFP activity. The EGFP, driven by the CMV promoter, was used as a positive control. The positive transformants showing green light of EGFP could be observed under fluorescent microscope. After G418 selection, the plasmid-transfected cell lines were obtained and the EGFP expression was measured using flow cytometry. In this study, the analysis of a series of the synchronized cell lines reveals the ProT a promoter is regulated by transcription factors and is independent on the cell cycle.