Ligase-based ultrasensitive detection of DNAzyme cleavage product using molecular beacon  

作　　者：Hong Peng Xie Xiang Xian Meng Hongyan Su Qing Yun Cai Yong Jun Tan Xiao Qin Huang 

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

出　　处：《Chinese Chemical Letters》2012年第10期1177-1180,共4页中国化学快报（英文版）

基　　金：supported in part by National Basic Research Program of China under Grants(No2009CB421601);Hunan Provincial Science and Technology Program(No2008SK3085)

摘　　要：Detection for deoxyribozyme （DNAzyme） cleavage usually needs complex and time-consuming radial labeling, gel electro- phoresis and autoradiography. A new approach was reported for detection DNAzyme cleavage product based on molecular beacon （MB）. Part of the loop of MB was designed to complementary to DNAzyme cleavage product. MB was employed to monitor ligation process of RNA/DNA complex and to convert directly cleavage product information into fluorescence signal. Detection limit of the assay is 0.02 nmol/L. The cleavage product of 8-17 DNAzyme against HCV-RNA was detected perfectly based on this assay. The method is fast, simple and ultrasensitive, which might hold great promise in DNAzyme reaction and DNAzyme gene therapy.Detection for deoxyribozyme （DNAzyme） cleavage usually needs complex and time-consuming radial labeling, gel electro- phoresis and autoradiography. A new approach was reported for detection DNAzyme cleavage product based on molecular beacon （MB）. Part of the loop of MB was designed to complementary to DNAzyme cleavage product. MB was employed to monitor ligation process of RNA/DNA complex and to convert directly cleavage product information into fluorescence signal. Detection limit of the assay is 0.02 nmol/L. The cleavage product of 8-17 DNAzyme against HCV-RNA was detected perfectly based on this assay. The method is fast, simple and ultrasensitive, which might hold great promise in DNAzyme reaction and DNAzyme gene therapy.

关 键 词：Molecular beacon LIGATION DNAzyme cleavage HCV-RNA 

分 类 号：Q523[生物学—生物化学] X831.02[环境科学与工程—环境工程]