Kinetic & Catalytic Activity Study of Lanthanide Complex of DO2A & K21DA in Phosphate Diester & DNA Hydrolysis

• Main Authors: Kuan, Pu-Yun, 管佈雲
• Other Authors: Chang C. Allen
• Format: Others
• Language: zh-TW
• Published: 1998
• Online Access: http://ndltd.ncl.edu.tw/handle/64199364745414106462

碩士 === 國立交通大學 === 生物科技研究所 === 86 === 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