mRNA detection in living cell using phosphorothioate-modified molecular beacon  

作　　者：TANG HongXing YANG XiaoHai WANG KeMin TAN WeiHong LI Wei 

机构地区：[1]State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Biomedical Engineering Center, and Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, China

出　　处：《Chinese Science Bulletin》2009年第9期1507-1514,共8页

基　　金：Supported by the National Key Basic Research Program (Grant No. 2002CB513110);International Technologies Collaboration Program of China (Grant No. 2003DF000039);National Natural Science Foundation of China (Grant Nos. 90606003 and 20620120107)

摘　　要：In this study, GFP mRNA in COS-7 cell and GFP-transfected COS-7 cell was detected in real time using phosphorothioate-modified molecular beacon based on living cell imaging method. Results showed that phosphorothioate-modified molecular beacon still kept the advantages of molecular beacon, such as, excellent selectivity, high sensitivity, and no separation detection. In addition, this modification could significantly increase the nuclease resistance of molecular beacon. Phosphorothioate-modified molecular beacon can efficiently reduce the false positive signal and improve the accuracy of living cell mRNA detection.In this study, GFP mRNA in COS-7 cell and GFP-transfected COS-7 cell was detected in real time using phosphorothioate-modified molecular beacon based on living cell imaging method. Results showed that phosphorothioate-modified molecular beacon still kept the advantages of molecular beacon, such as, excellent selectivity, high sensitivity, and no separation detection. In addition, this modification could significantly increase the nuclease resistance of molecular beacon. Phosphorothioate-modified molecular beacon can efficiently reduce the false positive signal and improve the accuracy of living cell mRNA detection.

关 键 词：分子信标 分离检测 MRNA 活细胞 灯塔 绿色荧光蛋白基因 ph 基因转染 

分 类 号：Q343[生物学—遗传学] TN247[电子电信—物理电子学]