Effects of metallic silver island films on resonance energy transfer between N,N′-(dipropyl)-tetra methyl indocarbocyanine (Cy3)- and N,N′-(dipropyl)-tetramethyl- indodicarbocyanine (Cy5)-labeled DNA

被引:45
|
作者
Malicka, J
Gryczynski, I
Kusba, J
Lakowicz, JR
机构
[1] Univ Maryland, Ctr Fluorescence Spect, Dept Biochem & Mol Biol, Baltimore, MD 21201 USA
[2] Gdansk Tech Univ, Fac Appl Phys & Math, PL-80952 Gdansk, Poland
关键词
N; '-(dipropyl)-tetramethylindocarbocyanine; '-(dipropyl)-tetramethylindodicarbocyanine; resonance energy transfer; silver island films;
D O I
10.1002/bip.10507
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Resonance energy transfer (RET) is typically limited to distances below 60 Angstrom, 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 Angstrom. 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. (C) 2003 Wiley Periodicals, Inc.
引用
收藏
页码:595 / 603
页数:9
相关论文
empty
未找到相关数据