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標題: | 阿拉伯芥中70kD熱休克蛋白質的分佈情形 Localization of 70 Kilodalton Heat Shock Protein in Arabidopsis thaliana |
作者: | 謝愷 |
出版年 : | 1995 |
學位: | 碩士 |
摘要: | 各種生物在遇到比正常生長溫度高五至十五度的熱逆境時,細胞會產生一些蛋白質,這群被熱所誘導而產生的蛋白質即稱為熱休克蛋白質。這群蛋白質中,以分子量為70kD的熱休克蛋白質(70kD Heat Shock Protein; HSP70)分佈最廣,也最為人所熟悉。 研究HSP70的功能時,HSP70在何種細胞內以及細胞中的何種位置顯得非常重要。本論文主要在以免疫標定法來觀察HSP70在細胞內部與組織間的分佈情形。 首先以純化過的兔子的抗阿拉伯芥70kD熱休克同質蛋白(70kD Heat Shock Cognate protein; HSC70)的抗體,做阿拉伯芥各個器官之蛋白質萃取液的免疫墨點法,證實了在常溫之下,阿拉伯芥的根、莖、葉、花、綠果莢、黃果莢,均有HSC70的存在。 因為曾經有報導用阿拉伯芥的HSC70 cDNA當探針對各個器官的RNA做過北方雜交法,發現在綠果莢、黃果莢中並無HSC70 mRNA存在,但免疫墨點法卻顯示有HSC70蛋白質存在。前後兩者的不一致,本文在此以反轉錄-聚合?連鎖反應來加以驗證。 最後則是分別以光學與電子顯微鏡來觀察常溫下HSC70在阿拉伯芥的組織與細胞中的位置,發現HSC70存在於植物細胞的細胞質與葉綠體中,並且大部分細胞都有表現,只是含量有所差異,本文並藉此推測HSC70在生物體內的功能。 Heat shock proteins are induced when the ambient temperature is elevated five to fifteen degrees Celsius in all biological systems. Among them, the 70 kilodalton heat shock proteins (HSP70) are the most widespread and the best known. The localization of HSP70 at the cell and tissue level is an important information in the study of the function of HSP70. In this research, the question is addressed using immunocytochemical techniques. Antibodies made against a 70 kilodalton heat shock cognate protein (HSC70) in Arabidopsis is used in immunoblot analysis. All organs of Arabidopsis, the root, stem, leaf, flower, developing silique as well as the mature silique are all shown to contain HSC70. It has been reported that in the siliques of various stages, mRNA of HSC70 was not detected based on RNA gel blot analysis. However, the immunoblot analysis confirmed the presence of HSC70-like proteins. This discrepancy is investigated here using reverse transcription-polymerase chain reaction. HSC70-like proteins are localized in Arabidopsis at the tissue and cell levels using light and electron microscopies, respectively. Almost all cells conatin HSC70, in the cytosol and the chloroplast, albeit varied in amount. The function of HSC70 is discussed based on this result. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76177 |
全文授權: | 未授權 |
顯示於系所單位: | 植物科學研究所 |
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