Sueda S, Yuan J, Matsumoto K
Department of Chemistry, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 169-8555, Japan.
Bioconjug Chem. 2000 Nov-Dec;11(6):827-31. doi: 10.1021/bc000030r.
Homogeneous DNA hybridization assay based on luminescence resonance energy transfer (LRET) from a tetradentate beta-diketonate europium chelate, 4,4'-bis(1' ',1' ',1' ',2' ',2' ',3' ',3' '-heptafluoro-4' ',6' '-hexanedion-6' '-yl)-chlorosulfo-o-terphenyl (BHHCT)-Eu(3+) (lambda(ex) = 340 nm and lambda(em) = 615 nm), to an organic dye, Cy5 (lambda(ex) = 643 nm and lambda(em) = 669 nm) has been developed, in which two DNA probes whose sequences comprises the whole complementary strand to the target DNA, are used; one probe having a biotin label on the 3'-terminus and the other a Cy5 label on the 5'-terminus. After hybridization, streptavidin labeled with BHHCT-Eu(3+) was added to the hybridization solution, and in the presence of the target DNA, the sensitized emission of Cy5 was observed when the hybridized complex was irradiated at 340 nm. In the absence of the target DNA, no emission was observed from Cy5.
基于四齿β-二酮铕螯合物4,4'-双(1'',1'',1'',2'',2'',3'',3''-七氟-4'',6''-己二酮-6''-基)-氯磺基邻三联苯（BHHCT）-Eu(3+)（激发波长λ(ex)=340nm，发射波长λ(em)=615nm）到有机染料Cy5（激发波长λ(ex)=643nm，发射波长λ(em)=669nm）的发光共振能量转移（LRET），开发了一种均匀DNA杂交分析方法。该方法使用了两个DNA探针，其序列包含与目标DNA的整个互补链；一个探针在3'-末端带有生物素标签，另一个在5'-末端带有Cy5标签。杂交后，将用BHHCT-Eu(3+)标记的链霉亲和素加入杂交溶液中，在目标DNA存在的情况下，当杂交复合物在340nm照射时，观察到Cy5的敏化发射。在没有目标DNA的情况下，未观察到Cy5的发射。
