Ultrasensitive monitoring of ribozyme cleavage product using molecular-beacon-ligation system  

作　　者：MENG XiangXian TANG ZhiWen WANG KeMin TAN WeiHong YANG XiaoHai LI Jun GUO QiuPing 

机构地区：[1]State Key Laboratory of Chemo/Biosensing and Chemometrics, Institute of Biological Technology, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China

出　　处：《Chinese Science Bulletin》2007年第5期603-607,共5页

基　　金：Supported by the National Key Basic Research Program (Grant No. 2002CB513100);the National Natural Science Foundation of China (Grant Nos. 20505007 and 20475015);the Key Project of Hunan Province Technology Plan of China (Grant Nos. 02JEY2004 and 0399Y1006)

摘　　要：This paper reports a new approach to detect ribozyme cleavage product based on the molecular-beacon-ligation system. The molecular beacon, designed in such a way that one-half of its loop is complementary to ribozyme cleavage product, is used to monitor ligation process of RNA/DNA com-plex in a homogeneous solution and to convert directly cleavage product information into fluorescence signal. The method need not label ribozyme and ribozyme substrate, which is fast, simple and ultra-sensitive for detection of cleavage product. Detection limit of the assay is 0.05 nmol/L. The cleavage product of hammerhead ribozyme against hepatitis C virus RNA (HCV-RNA) was detected perfectly based on this assay. Owing to its ultrasensitivity, excellent specificity, convenience and fidelity, this method might hold out great promise in ribozyme reaction and ribozyme gene therapy.This paper reports a new approach to detect ribozyme cleavage product based on the molecularbeacon-Iigation system. The molecular beacon, designed in such a way that one-half of its loop is complementary to ribozyme cleavage product, is used to monitor Iigation process of RNA/DNA complex in a homogeneous solution and to convert directly cleavage product information into fluorescence signal. The method need not label ribozyme and ribozyme substrate, which is fast, simple and ultrasensitive for detection of cleavage product. Detection limit of the assay is 0.05 nmol/L. The cleavage product of hammerhead ribozyme against hepatitis C virus RNA （HCV-RNA） was detected perfectly based on this assay. Owing to its ultrasensitivity, excellent specificity, convenience and fidelity, this method might hold out great promise in ribozyme reaction and ribozyme gene therapy.

关 键 词：核酶 分裂产物 核酸连接 分子信标连接系统 超灵敏监测 

分 类 号：Q556.5[生物学—生物化学]