Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/20898
標題: 小球藻Chlorella sp. strain DT種和211-8b種於乾燥逆境下可溶性蛋白 圖譜之變化
Changes in soluble protein complements in dehydration of the desiccation-tolerant Chlorella sp. stain DT and the desiccation- sensitive Chlorella pyrenoidosa 211-8b
作者: 彭雨文
Peng, Yu-Wen
關鍵字: Chlorella
小球藻
dehydration
soluble protein
2-D electrophoresis
乾燥
可溶性蛋白質
雙向電泳
出版社: 植物學系
摘要: 本實驗室分離的小球藻strain DT與取自德國藻類中心的小球藻strain 211-8b,以500×104cells/ml起始培養兩天之後作為材料。利用紅外線輻 射加熱天平以42℃加熱,約在3小時後幾近完全乾燥,以60℃加熱則在130 分鐘後接近完全乾燥。乾燥的細胞在相對濕度25% 的乾燥箱中貯存三天以 上,重新以液體培養基培養,除了42℃烘乾的DT可於七至八天恢復生長外 ,DT以60℃烘乾或8b以42℃、60℃烘乾後皆無法重新恢復生長。以5% ∼15% native線性梯度聚丙烯醯胺膠體電泳,比較新鮮DT細胞和42℃乾燥 後蛋白質之變化,發現乾燥細胞於分子量250kD處有明顯的可溶性蛋白質 新合成,並隨著乾燥時間的增長有合成量增多的趨勢。將新鮮與42℃乾燥 的DT細胞,以不同濃度的飽和硫酸銨進行蛋白質沈澱分割,進行電泳分析 ,發現大部分的蛋白質在20%∼40% 和40%∼60% 的片段中被沈澱下來,而 乾燥細胞中新發現的250kD蛋白質在硫酸銨沈澱處理後消失。而以SDS- PAGE分析新鮮與42℃烘乾的DT和8b細胞,其可溶性蛋白質的種類增多,但 不易比較其中之差異。DT新鮮細胞的可溶性蛋白質在雙向電泳上分佈之圖 譜,大約可以偵測到280種上的蛋白質;同種的藻細胞在不同處理期間基 本圖譜大致相似,然而量或質的改變存在於不同處理組間。相較於新鮮的 DT細胞,42℃烘乾後共有29種蛋白質含量增加或新合成,26種減少或消失 ;8b含量上升的有21種,下降的共有37種。其中有三種是僅在DT發現的新 合成蛋白質:DT-1 (40.87kD), DT-14 (51.08kD)和DT-19 (51.08kD), 其分子量恰為native-PAGE所發現耐乾燥蛋白質分子量的1/5。DT恢復培養 以二天為間隔萃取細胞中可溶性蛋白質,進行雙向電泳圖譜,在恢復培養 二天及四天,蛋白質的量及種類皆非常少,變化很大,呈極不穩定狀態。 恢復培養六天後,蛋白質的種類與含量顯著增多,到第七天藻細胞的蛋白 質雙向電泳圖譜才近似新鮮的可溶性蛋白質圖譜。
Desiccation-tolerant Chlorella sp. strain DT(from an arid mountainous area of Taiwan) and Desiccation-sensitive Chlorella pyrenoidosa (211-8b, from the AlgaeCollection Center, Univ, Gottingen, Germany) were cultured Chlorella-medium.Only a limited amount of water can be conserved in the DT and 8b cell when dehydrate using an infra-red radiation balance at 42oC for 3 h, and it tookonly 130min to dehydrate the Chlorella cells completely when heated at 60oC. The dried cells stored in desiccation box for 3 days, the DT cells dehydrate 42oC could restore growth absolutely after 7-8 days rewetting in liquid culturemedium. The DT dehydrate at 60oC and the 8b dehydrate at 42oC or 60oC could not restore growth.Two-dimensional gel electrophoresis analysis of polypeptides revealed that thepatterans of fresh and dehydrated at 42oC differed significantly in their polypeptide profiles. As shown in the figure, approximately 280 polypeptides could be resolved by the 2-D system. The changes included increase, decrease ornewly induced synthesis. In general, labelling patterns were similar despite sample differences.The difference became conspicuously after desiccation DTrestored growth in liquid culture medium.
URI: http://hdl.handle.net/11455/20898
Appears in Collections:生命科學系所

文件中的檔案:

取得全文請前往華藝線上圖書館



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.