Synthesis of a fluorescent xanthenic derivative useful for labeling amine residues.
The 2,5-dioxopyrrolidin-1-yl-4-(3-hydroxy-6-oxo-6H-xanthen-9-yl)-3-methylbenzoate has been synthesized as an amine-reactive derivative able to yield stable covalently labeled biopolymers. The new derivative has been used to label polyribocytidilic acid (5'), poly(C), amine residues. TG-II-poly(C) exhibits monoexponential decay at the physiological pH range. In addition, both steady-state fluorescence intensity and fluorescence decay are also sensitive to solution pH. The large decrease in steady-state fluorescence upon hybridization allows it to be used as a nucleic acid probe in a homogeneous assay format. In summary, we report an efficient synthesis to obtain labeled RNA from commercially available materials in excellent yields.
Crovetto
Luis
L
Rios
Ramon
R
Alvarez-Pez
Jose M
JM
Paredes
Jose M
JM
Lozano-Velez
Patricia
P
del Valle
Carmen
C
Talavera
Eva M
EM
D000588:Amines / Q000737:chemistry*; D000838:Anions; D002627:Chemistry, Physical / Q000379:methods; D002845:Chromatography / Q000379:methods; D004396:Coloring Agents / Q000737:chemistry*; D019793:Fluorescein / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008956:Models, Chemical; D008962:Models, Theoretical; D015345:Oligonucleotide Probes / Q000737:chemistry; D012313:RNA / Q000737:chemistry; D014966:Xanthenes / Q000737:chemistry*
D000588:Amines; D000838:Anions; D004396:Coloring Agents; D015345:Oligonucleotide Probes; D014966:Xanthenes; D012313:RNA; D019793:Fluorescein