Photoregulation of DNA polymerase I (Klenow) with caged fluorescent oligodeoxynucleotides.
The DNA polymerase reaction by Klenow fragment (KF) was efficiently regulated with UV light using a 25-mer caged fluorescent oligodeoxynucleotide (CFO) as the template. The CFO was functionalized with a fluorescein reporter (Fl) and photocleavable DABSYL quencher moiety (Dab). With Fl and Dab at adjacent cytidines in the middle at the template, KF was blocked from extending a complementary 12-mer primer. Upon UV photolysis of the DABSYL blocking group under aerobic conditions, fluorescein emission was restored and 50% of the primers were fully extended by KF.
Tang
XinJing
X
Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, PA 19104-6323, USA.
Richards
Julia L
JL
Peritz
Adam E
AE
Dmochowski
Ivan J
IJ
D004256:DNA Polymerase I / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000378:metabolism / Q000528:radiation effects*; D010782:Photolysis; D013698:Templates, Genetic; D014466:Ultraviolet Rays
D005456:Fluorescent Dyes; D009838:Oligodeoxyribonucleotides; D004256:DNA Polymerase I