Real-time fluorescence detection of exoribonucleases.
The identification of RNases or RNase effectors is a continuous challenge, particularly given the current importance of RNAs in the control of genome expression. Here, we show that a fluorogenic RNA-DNA hybrid is a powerful tool for a real-time fluorescence detection and assay of exoribonucleases (RT-FeDEx). This RT-FeDEx assay provides a new strategy for the isolation, purification, and assay of known and unknown exoribonucleases.
Sinturel
Flore
F
Institut de Biologie Physico-Chimique, CNRS UPR 9073, Université de Paris 7-Denis Diderot, 75005 Paris, France.
Pellegrini
Olivier
O
Xiang
Song
S
Tong
Liang
L
Condon
Ciarán
C
Bénard
Lionel
L
D001483:Base Sequence; D005095:Exoribonucleases / Q000032:analysis* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000032:analysis*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013997:Time Factors
D005456:Fluorescent Dyes; D005095:Exoribonucleases