Two new photolabile nucleotide analogues with furan-fused deoxyuridine were synthesized through Sonogashira coupling.Their enzymatic incorporation into DNA was evaluated with two DNA polymerases(Taq and Deep vent exo-...Two new photolabile nucleotide analogues with furan-fused deoxyuridine were synthesized through Sonogashira coupling.Their enzymatic incorporation into DNA was evaluated with two DNA polymerases(Taq and Deep vent exo-)by polymerase chain reaction(PCR).Deep vent exo-recognized both nucleotides as substrates for primer extension,while Taq was much less proficient.Light irradiation of PCR products released the amino and carboxyl moieties of DNA.Further labeling with fluorescein isothiocyanate for a long DNA construct with F-dUnTP incorporation was successfully achieved.展开更多
基金supported by the National Natural Science Foundation of China(21072015)the National Basic Research Program of China(973 Program,2012CB720600)+1 种基金the Program for New Century Excellent Talents in University(NCET-10-0203)the State Key Laboratory of Drug Research(SIMM1106KF-15)
文摘Two new photolabile nucleotide analogues with furan-fused deoxyuridine were synthesized through Sonogashira coupling.Their enzymatic incorporation into DNA was evaluated with two DNA polymerases(Taq and Deep vent exo-)by polymerase chain reaction(PCR).Deep vent exo-recognized both nucleotides as substrates for primer extension,while Taq was much less proficient.Light irradiation of PCR products released the amino and carboxyl moieties of DNA.Further labeling with fluorescein isothiocyanate for a long DNA construct with F-dUnTP incorporation was successfully achieved.
