Kinetics studies of lanthanide polyaza polycarboxylate macrocyclic complexes Ln(DO2A) and their applications in DNA/RNA hydrolytic scission

• Main Author: 陳家翎
• Other Authors: 張正
• Format: Others
• Language: zh-TW
• Published: 2000
• Online Access: http://ndltd.ncl.edu.tw/handle/47863230735046783905

碩士 === 國立交通大學 === 生物科技研究所 === 88 === ENGLISH ABSTRACT Since the first wholly synthetic artificial ribozyme was published in 1994, DNA and RNA whose half-life is very long can also be degradation in the short time. Therefore, it took scientists’ attention and researches, and is to promote the DNA’s or RNA’s hydrolysis rate became a challenge for many biochemists. Because the artificial restriction enzyme may be applied for gene engineering and probably provided a more effective and harmless cure. In the beginning of this thesis, we are using macrocyclic ligand 1,7-bis(carboxymethyl)1,4,7,10-tetraazacyclododecane (DO2A) and lanthanide ions to from complexes, and study their dissociation kinetics. Based on the obtained rate constants, we have found that, DO2A have strong binding affinity with lanthanide ions. Moreover, we proved that these complexes have very high stabilities again. In the other side, we use Ce(DO2A)+ and Eu(DO2A)+ to cleavage double-strand DNA(pUC19) and obtained the value of kcat are 1.0601×10-3, 3.426×10-4(s-1) and Km are 1.785×10-3,9.13210×-4(M). Further, we use Eu(DO2A)+ to hydrolysis the phosphate triester bonds at the cap structure of the mRNA. Also, we achieved the kcat which is 1.746×10-4(s-1). Compared with other’s data, it proved that the complexes have the excellent capability to cleave DNA and RNA.