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標題: 應用金黃葡萄球菌鎘離子運輸蛋白CadA於阿拉伯芥重金屬抗性之研究
Application of the Staphylococcus aureus cadmium-transporter CadA in Arabidopsis thaliana for heavy metal resistance
作者: 陳君如
Chen, Jiun-Ru
關鍵字: Arabidopsis thaliana

Staphylococcus aureus
出版社: 生命科學系所
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摘要: 鎘和鉛在工業化國家中被廣泛地利用,但對於生物體而言,鎘和鉛是非必需重金屬元素,一但累積在生物體內,則會對生物造成毒害。曾在日本發生因汞中毒所引起的水俁病(Minamata disease),鎘中毒所引起的痛痛病(Itai-itai disease),都是因為在食物鏈中生物的累積性,使得微量的污染也深具威脅性,因此,將非必需重金屬累積程度減至最小之機制是所有生物體所需要的。本研究主要目的是利用植物基因轉殖工程將已知的革蘭氏陽性菌金黃葡萄球菌(Staphylococcus aureus)質體pI258上的抗鎘系統(cadCA operon)中之鎘離子運輸蛋白的cadA基因轉殖至阿拉伯芥(Arabidopsis thaliana)中,使其在植物細胞內表現,來探討cadA基因的轉殖植物對鎘、鉛或鋅能否增加抗性及減少累積量,以利日後作為降低累積性或植生復育(Phytoremediation)的工具。本實驗是以帶有花椰菜嵌紋病毒CaMV 35S啟動子的載體pBI121來推動cadA基因在阿拉伯芥內表現,利用農桿菌轉殖法(Agrobacterium mediated transformation)將CadA基因嵌入阿拉伯芥基因體中。種子每代皆以抗生素Kanamycin作篩選,而得到帶有CadA基因的第三子代同型接合(homozygous)的轉殖株種子。利用聚合酶連鎖反應和反轉錄聚合酶連鎖反應得知cadA基因確實有嵌入阿拉伯芥基因體中且有表現。在含鎘的耐受性發芽測試結果顯示,和野生型阿拉伯芥相比較之下,表現cadA基因的轉殖植物對鎘的抗性確實提高。另外,在含鉛或鋅的抗性測試結果顯示,和野生型阿拉伯芥相比較之下,表現cadA基因的轉殖植物對鉛或鋅的抗性確實提高。但在鎘及鉛累積量的檢測中,表現cadA基因的轉殖植物對鎘或鉛的累積量卻都只比野生型阿拉伯芥的累積量微高些。本研究證實利用基因工程的方法將cadA基因表現於植物中的確能提高植物對鎘、鉛及鋅的耐受性,未來可應用在使受重金屬污染土地中之植物增加對重金屬的抗性,以作為植生復育(Phytoremediation)的工具。
Cadmium(Cd) and lead(Pb) are widely used in industrialized countries, and both are non-essential heavy metals for the organisms. When they are accumulated, they will become extremely toxic to living organisms. For instance, the Minimata disease caused by mercury contamination, and the Itai-itai disease caused by cadmium contamination had made serious injury to the biological system for its bioaccumulation in the food chain. These heavy metals could be a threat at the low levels. Therefore, mechanisms responsible for minimizing the concentraction of non-essential heavy metals is required for all organisms. This study was initiated to clone and characterise of cadA gene originated from cadCA operon in a gram-positive bacteial plasmid, Staphylococcus aureus plasmid pI258, into Arabidopsis thaliana.The expression of cadA gene in A. thaliana may increase its resistance to cadmium, lead and zinc(Zn) and decrease the heavy metal content in the transgenic plants. The isolated transgenic lines were conferred both in DNA and RNA levels. Analysis of transgenic A. thaliana plants expressing cadA showed that CadA is functionally active and that the plants have enhanced resistance of Cd(II), Pb(II) and Zn(II), while accumulated a greater amounts of Cd(II) or Pb(II). These results suggest that transgenic plants expressing cadA may be an useful tool for phytoremediation.
其他識別: U0005-2507200610364500
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