Novel application of Phi29 DNA polymerase: RNA detection and analysis in vitro and in situ by target RNA-primed RCA

被引：38

作者：

Lagunavicius, Arunas ^{[1
]}

Merkiene, Egle ^{[1
]}

Kiveryte, Zivile ^{[1
]}

Savaneviciute, Agne ^{[1
]}

Zimbaite-Ruskuliene, Vilma ^{[1
]}

Radzvilavicius, Tomas ^{[1
]}

Janulaitis, Arvydas ^{[1
]}

机构：

[1] Fermentas UAB, LT-02241 Vilnius, Lithuania

来源：

RNA | 2009年 / 15卷 / 05期

关键词：

exoribonuclease; GAPDH; Phi29 DNA polymerase; RCA; RNA analysis; RNase; ROLLING-CIRCLE AMPLIFICATION; PADLOCK PROBES; MESSENGER-RNA; SINGLE CELLS; MOLECULES; SEQUENCES; LIGATION;

D O I：

10.1261/rna.1279909

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We present a novel Phi29 DNA polymerase application in RCA-based target RNA detection and analysis. The 3'-> 5' RNase activity of Phi29 DNA polymerase converts target RNA into a primer and the polymerase uses this newly generated primer for RCA initiation. Therefore, using target RNA-primed RCA, padlock probes may be targeted to inner RNA sequences and their peculiarities can be analyzed directly. We demonstrate that the exoribonucleolytic activity of Phi29 DNA polymerase can be successfully applied in vitro and in situ. These findings expand the potential for detection and analysis of RNA sequences distanced from 3'-end.

引用

页码：765 / 771

页数：7