Homogeneous DNA hybridization assay by using europium luminescence energy transfer
Homogeneous DNA hybridization assay by using europium luminescence energy transfer
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DOI：
10.1021/bc000030r
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发表时间：
2000-11-01
影响因子：
4.7
通讯作者：
Matsumoto, K
中科院分区：
文献类型：
--
作者：
Sueda, S;Yuan, JL;Matsumoto, K
4，4 ′-双四齿β-二酮铕螯合物的荧光共振能量转移（LRET）-DNA均相杂交分析（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 +（λ（ex）= 340 nm和λ（em）= 615 nm）与有机染料Cy 5（λ（em）= 643 nm和λ（em）= 669 nm），其中使用了两种DNA探针，其序列包含靶DNA的整个互补链;一个探针在3 ′-末端具有生物素标记，另一个在5 ′-末端具有Cy 5标记。杂交后，将BHHCT-Eu 3+标记的链霉亲和素添加到杂交溶液中，并且在靶DNA的存在下，当在340 nm下照射杂交复合物时，观察到Cy 5的敏化发射。在不存在靶DNA的情况下，未观察到Cy 5的发射。
Homogeneous DNA hybridization assay based on luminescence resonance energy transfer (LRET) from st 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)-Eu3+ (lambda (ex) = 340 nm and lambda (em) = 615 nm), to an organic dye, Cy5 (lambda (em) = 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-Eu3+ 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.