Malicka Joanna, Gryczynski Ignacy, Kusba Jozef, Lakowicz Joseph R
University of Maryland Baltimore, Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201, USA.
Biopolymers. 2003 Dec;70(4):595-603. doi: 10.1002/bip.10507.
Resonance energy transfer (RET) is typically limited to distances below 60 A, which can be too short for some biomedical assays. We examined a new method for increasing the RET distances by placing donor- and acceptor-labeled DNA oligomers between two slides coated with metallic silver particles. A N,N'-(dipropyl)-tetramethylindocarbocyanine donor and a N,N'-(dipropyl)-tetramethylindodicarbocyanine acceptor were covalently bound to opposite 5' ends of complementary 23 base pair DNA oligomers. The transfer efficiency was 25% in the absence of silver particles or if only one slide was silvered, and it increased to an average value near 64% between two silvered slides. The average value of the Forster distance increased from 58 to 77 A. The energy transfer data were analyzed with a model assuming two populations of donor-acceptor pairs: unaffected and affected by silver island films. In an affected fraction of about 28%, the apparent energy transfer efficiency is near 87% and the Forster distance increases to 119 A. These results suggest the use of metallic silver particles to increase the distances over which RET occurs in biomedical and biotechnology assays.
共振能量转移(RET)通常限于60埃以下的距离,这对于某些生物医学检测来说可能太短。我们研究了一种新方法,通过在两片涂有金属银颗粒的载玻片之间放置供体和受体标记的DNA寡聚物来增加RET距离。一个N,N'-(二丙基)-四甲基吲哚碳菁供体和一个N,N'-(二丙基)-四甲基吲哚二碳菁受体共价连接到互补的23个碱基对DNA寡聚物的相对5'端。在没有银颗粒的情况下或仅一片载玻片镀银时,转移效率为25%,而在两片镀银载玻片之间,转移效率增加到平均值接近64%。福斯特距离的平均值从58埃增加到77埃。用一个假设供体-受体对有两种群体的模型分析了能量转移数据:不受银岛膜影响的和受银岛膜影响的。在约28%的受影响部分中,表观能量转移效率接近87%,福斯特距离增加到119埃。这些结果表明,在生物医学和生物技术检测中可利用金属银颗粒来增加RET发生的距离。