Inhibition of Ampicillin-resistance in Bacteria by Modified DNAzymes  

作　　者：CHEN Fei LI Zhe YANG Shuo WANG Rui-jian LIU Bin SONG Yu-ming SUN Yan-hong HAO Dong-yun WANG Xiao-ping 

机构地区：[1]Key Lab for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130021, P. R. China [2]China-Japan Friendship Hospital, Jilin University, Changchun 130031, P. R. China

出　　处：《Chemical Research in Chinese Universities》2008年第4期491-495,共5页高等学校化学研究（英文版）

基　　金：the National Natural Science Foundation of China(Nos.20771030 and 20671025)

摘　　要：To overcome ampicillin-resistance of bacteria which is believed to attribute their endogenous B-lactamase, we designed three 10-23 DNAzymes（Dz1, Dz2. Dz3） targeting the coding region of B-lactamase mRNA and examined their inhibitory capabilities of the ampicillin-resistance of TEM-1 and TEM-3 bacteria. Dz1 was a traditional 10-23 DNAzyme, Dz2 was the mutant of Dz1 by addition of the protected nucleotide to each ann of the enzyme, and Dz3 was a mutant of Dz1 at antisense arms of which phosphorothioate modifications were made. Kinetic analysis, bacterial growth, and β-lactamase activity measurement showed that all the three DNAzymes worked efficiently in vitro and in vivo. A 9 hours bacterial growth inhibition test showed that the inhibition rates of TEM-1 bacteria by Dz1, Dz2, and Dz3 were 27%, 50%, and 29%, respectively. In addition, the inhibition rates of TEM-3 bacteria by those three DNAzymes were found io be 49%, 58%, and 45%, respectively. The current findings suggest that DNAzymes may become potential candidates of alternative inhibitors for bacteria drug-resistance.To overcome ampicillin-resistance of bacteria which is believed to attribute their endogenous B-lactamase, we designed three 10-23 DNAzymes（Dz1, Dz2. Dz3） targeting the coding region of B-lactamase mRNA and examined their inhibitory capabilities of the ampicillin-resistance of TEM-1 and TEM-3 bacteria. Dz1 was a traditional 10-23 DNAzyme, Dz2 was the mutant of Dz1 by addition of the protected nucleotide to each ann of the enzyme, and Dz3 was a mutant of Dz1 at antisense arms of which phosphorothioate modifications were made. Kinetic analysis, bacterial growth, and β-lactamase activity measurement showed that all the three DNAzymes worked efficiently in vitro and in vivo. A 9 hours bacterial growth inhibition test showed that the inhibition rates of TEM-1 bacteria by Dz1, Dz2, and Dz3 were 27%, 50%, and 29%, respectively. In addition, the inhibition rates of TEM-3 bacteria by those three DNAzymes were found io be 49%, 58%, and 45%, respectively. The current findings suggest that DNAzymes may become potential candidates of alternative inhibitors for bacteria drug-resistance.

关 键 词：DNAZYME DRUG-RESISTANCE Β-LACTAMASE AMPICILLIN Antisense drug 

分 类 号：R978.1[医药卫生—药品]