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吳郭魚腦細胞株TB2部分間隙蛋白43基因的選殖 = cloning of...

國立高雄大學生物科技研究所

吳郭魚腦細胞株TB2部分間隙蛋白43基因的選殖 = cloning of partial connexin43 from TB2 derived from tilapia brain

紀錄類型:	書目-語言資料,印刷品 : 單行本
並列題名:	cloning of partial connexin43 from TB2 derived from tilapia brain
作者:	黃馨儀,
其他團體作者:	國立高雄大學
出版地:	[高雄市]
出版者:	撰者;
出版年:	2008[民97]
面頁冊數:	66面圖，表 : 30公分;
標題:	星狀膠細胞
標題:	astrocyte
電子資源:	http://handle.ncl.edu.tw/11296/ndltd/79930464910510726714
附註:	指導教授：溫秋明
附註:	參考書目：面47-52
附註:	附錄：實驗藥品等4種
摘要註:	間隙連接在相鄰細胞的細胞膜中形成可以讓離子和代謝物質通過的通道，此通道稱為connexon，由6個稱為間隙蛋白（connexin, Cx）的穿膜蛋白質組成。間隙連接在中樞神經系統扮演重要角色，各種類的神經細胞有特定的Cx。間隙連接的表現不僅會影響中樞神經細胞的功能，也和其發育和分化有關。在中樞神經系統中，室管膜細胞和星狀膠細胞及其前驅細胞的主要間隙連接由Cx43構成。吳郭魚的神經原始細胞（TB2）已被證實具有Cx43，但其基因和蛋白質組成尚未清楚，表現調控因子也還不明瞭。為探討TB2細胞之Cx43基因是否會受分化因子影響其表現，本論文以數種濃度的視黃酸及多種時間處理TB2細胞，以半定量RT-PCR方式檢測Cx43 mRNA的表現。結果顯示視黃酸會促進TB2細胞 Cx43 RNA表現。由於商品化之Cx43抗體和多種TB2細胞的蛋白質反應，無法用來分析TB2細胞的Cx43，因而自行開發魚類Cx43專一性抗體。利用重組蛋白方式，本論文開發出大鼠抗TB2 Cx43抗體，並利用此抗體以西方點墨分析TB2細胞的 Cx43蛋白質，結果在約76kD有明顯條帶，和哺乳動物在43kD的Cx43明顯不同。因此，本論文也進行選殖分析TB2細胞的Cx43 mRNA。另外，利用重組大鼠抗TB2細胞 Cx43重組蛋白抗體，本論文探討和TB2 細胞中可以和Cx43作用的蛋白質。 Gap junctions are cell-to-cell channels allowing direct diffusion of ions and metabolites . Several biological functions have been proposed, such as involvement in embryonic development, cell differentiation and growth control. Each channel is formed by docking and opening of hemichannels in adjacent cells consisting of connexin (Cx).Cx43 is major gap-junction subunit protein in ependymal cells and astrocytes as well as the precursor cells. Putative neural progenitor cell derived from tilapia brain (TB2) has verified exhibiting Cx43. But Cx43 gene and protein make up not yet clear, it is not still clear to control of regulatory factor. Whether will be influenced it to Cx43 expression of TB2 cell by differentiation factor, this paper deals with TB2 cell with several kinds of concentration of retinoic acid and time to measure the expression of Cx43 mRNA by half-quantitative RT-PCR. The result that Cx43 RNA level in TB2 cell is upregulated by retinoic acid. Because commercial Cx43 antibody is interact with many protein of TB2 cell, it is unable to be used for analysing Cx43 of TB2 cell. Therefore develop the fish Cx43 antibody by oneself. Utilize recombination protein to develops the rat anti-TB2 Cx43 antibody, and the result of western blot find Cx43 protein in TB2 cell has remarkable band at 76kD, obviously different in Cx43 of 43kD in mammals. So this paper used cloning to analyse Cx43 mRNA of TB2 cell. In addition, is it recombination rat anti-TB2 Cx43 recombination protein antibody can be interact with proteins of TB2 cell.

吳郭魚腦細胞株TB2部分間隙蛋白43基因的選殖 = cloning of partial connexin43 from TB2 derived from tilapia brain
黃, 馨儀

吳郭魚腦細胞株TB2部分間隙蛋白43基因的選殖 = cloning of partial connexin43 from TB2 derived from tilapia brain / 黃馨儀撰 - [高雄市] : 撰者, 2008[民97]. - 66面 ; 圖，表 ; 30公分.
指導教授：溫秋明參考書目：面47-52附錄：實驗藥品等4種.
星狀膠細胞astrocyte

吳郭魚腦細胞株TB2部分間隙蛋白43基因的選殖 = cloning of partial connexin43 from TB2 derived from tilapia brain
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330 $a 間隙連接在相鄰細胞的細胞膜中形成可以讓離子和代謝物質通過的通道，此通道稱為connexon，由6個稱為間隙蛋白（connexin, Cx）的穿膜蛋白質組成。間隙連接在中樞神經系統扮演重要角色，各種類的神經細胞有特定的Cx。間隙連接的表現不僅會影響中樞神經細胞的功能，也和其發育和分化有關。在中樞神經系統中，室管膜細胞和星狀膠細胞及其前驅細胞的主要間隙連接由Cx43構成。吳郭魚的神經原始細胞（TB2）已被證實具有Cx43，但其基因和蛋白質組成尚未清楚，表現調控因子也還不明瞭。為探討TB2細胞之Cx43基因是否會受分化因子影響其表現，本論文以數種濃度的視黃酸及多種時間處理TB2細胞，以半定量RT-PCR方式檢測Cx43 mRNA的表現。結果顯示視黃酸會促進TB2細胞 Cx43 RNA表現。由於商品化之Cx43抗體和多種TB2細胞的蛋白質反應，無法用來分析TB2細胞的Cx43，因而自行開發魚類Cx43專一性抗體。利用重組蛋白方式，本論文開發出大鼠抗TB2 Cx43抗體，並利用此抗體以西方點墨分析TB2細胞的 Cx43蛋白質，結果在約76kD有明顯條帶，和哺乳動物在43kD的Cx43明顯不同。因此，本論文也進行選殖分析TB2細胞的Cx43 mRNA。另外，利用重組大鼠抗TB2細胞 Cx43重組蛋白抗體，本論文探討和TB2 細胞中可以和Cx43作用的蛋白質。 Gap junctions are cell-to-cell channels allowing direct diffusion of ions and metabolites . Several biological functions have been proposed, such as involvement in embryonic development, cell differentiation and growth control. Each channel is formed by docking and opening of hemichannels in adjacent cells consisting of connexin (Cx).Cx43 is major gap-junction subunit protein in ependymal cells and astrocytes as well as the precursor cells. Putative neural progenitor cell derived from tilapia brain (TB2) has verified exhibiting Cx43. But Cx43 gene and protein make up not yet clear, it is not still clear to control of regulatory factor. Whether will be influenced it to Cx43 expression of TB2 cell by differentiation factor, this paper deals with TB2 cell with several kinds of concentration of retinoic acid and time to measure the expression of Cx43 mRNA by half-quantitative RT-PCR. The result that Cx43 RNA level in TB2 cell is upregulated by retinoic acid. Because commercial Cx43 antibody is interact with many protein of TB2 cell, it is unable to be used for analysing Cx43 of TB2 cell. Therefore develop the fish Cx43 antibody by oneself. Utilize recombination protein to develops the rat anti-TB2 Cx43 antibody, and the result of western blot find Cx43 protein in TB2 cell has remarkable band at 76kD, obviously different in Cx43 of 43kD in mammals. So this paper used cloning to analyse Cx43 mRNA of TB2 cell. In addition, is it recombination rat anti-TB2 Cx43 recombination protein antibody can be interact with proteins of TB2 cell.
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