標題: 鑭係金屬與大環配位子錯合物切割磷酸二酯與去氧核醣核酸動力學及催化效力的研究
Kinetic & Catalytic Activity Study of Lanthanide Complex of DO2A & K21DA in Phosphate Diester & DNA Hydrolysis
作者: 管佈雲
Kuan, Pu-Yun
張 正
Chang C. Allen
生物科技學系
關鍵字: 大環配位子錯合物;Lanthanide Complex
公開日期: 1997
摘要: 去氧核醣核酸(DNA) 在自然的狀態下非常穩定,其半生期超過一億年,因 此,加速DNA 的水解速率便成為許多生化學家的挑戰。在本篇論文中,我 們利用大環配位子DO2A (1,7-bis(carboxymethyl) 1,4,7,10-tetraazacyclododecane) 及K21DA (1,7-diaza-4,10,13- trioxacyclopentadecane N,N'-diacetic acid) 和鑭系金屬離子La(III) 、Eu(III)形成錯合物。利用金屬離子上配位水數目的不同,計算錯合物 切割磷酸二酯(BNP-) 的速率。發現,EuDO2A 有相當不錯的切割效率。其 反應常數(pH=8, k'=1.23’102) 為LaDO2A 之兩倍(pH=8.05, k'=0.574 ’102)及EuK21DA的一百五十倍之後,我們選擇EuDO2A切割單股以及雙股 的去氧核醣核酸, 首先利用HPLC 觀測EuDO2A 水解單股DNA Primer 的效 力,發現反應兩小時後,DNA primer 量減少並且有核酸的碎片出現。五 小時後二十六個鹼基的DNA Primer 完全被水解。最後,用Agarose 電泳 的方式觀測EuDO2A 水解雙股DNA 的結果。發現DNA Form I 隨錯合物的量 增加而漸減;而Form II 卻隨錯合物的量增加而漸增,證明EuDO2A 是有 切割DNA 的效力。 Deoxyribonucleic acid (DNA) at the natural state is very stable; its half-life is over 100,000,000 years. Therefore, how to promote the DNA's hydrolysis rate becomes a challenge for many biochemists. In the beginning of this thesis, we are using macrocyclic ligand 1,7-bis(carboxymethyl) 1,4,7,10-tetraazacyclododecane (DO2A) and acid-1,7- diaza-4,10,13-trioxacyclopentadecane N,N'-Diacetic (K21DA) and lanthanide ions La(III) and Eu(III) to form complexes, and cleavage the phosphate diester(BNP-). By using diffFor the cleavage of the deoxyribonucleic acid, we used the HPLC to observe the hydrolysis of single-strand DNA primer by EuDO2A. After 5 hours, the 26 base DNA primer was completely hydrolyzed. We also used the agarose gel electrophoresis to observe the result of hydrolysis of double-strand DNA.It was found that DNA Form I (supercoiled form) decreased as the concentration of the complex increased, and DNA Form II (circular form) increased as the concentration of the complex increased. This proves that the E
URI: http://140.113.39.130/cdrfb3/record/nctu/#NT860111011
http://hdl.handle.net/11536/62590
顯示於類別:畢業論文