Share Email Print
cover

Proceedings Paper

Single RNA kissing complexes studied by fluorescence resonance energy transfer
Author(s): Peter B. Yim; Xiaoyi Zhang; Eric S. DeJong; Jennifer M. Carroll; John P. Marino; Lori S. Goldner
Format Member Price Non-Member Price
PDF $14.40 $18.00

Paper Abstract

We have used single molecular-pair fluorescence resonance energy transfer (FRET) to probe the conformation of a RNA loop-loop “kissing complex” formed by two small RNA hairpins (R1inv and R2inv) derived from Escherichia coli (ColE1) plasmid-encoded transcripts, RNA I and RNA II. This RNA kissing complex is a critical intermediate in a multi-step hybridization pathway which controls plasmid replication. Biotinylated RNA molecules were labeled with donor and acceptor dyes on their 5' ends and immobilized on a biotinylated surface using streptavidin. Fluorescence from the donor and acceptor dyes was collected and measured by photon counting detectors in two spectrally separated channels in a customized confocal microscope. Quantitative measurement of intramolecular distances between 5' ends of the RNA was obtained using donor-only single molecule FRET. This donor-only single molecule FRET technique is described in detail and validated through determination of the distance between 5' ends of 8mer A-form RNA helices of known structure.

Paper Details

Date Published: 29 March 2005
PDF: 11 pages
Proc. SPIE 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III, (29 March 2005); doi: 10.1117/12.590896
Show Author Affiliations
Peter B. Yim, National Institute of Standards and Technology (United States)
Xiaoyi Zhang, National Institute of Standards and Technology (United States)
Eric S. DeJong, Ctr. for Advanced Research in Biotechnology (United States)
Jennifer M. Carroll, National Institute of Standards and Technology (United States)
John P. Marino, Ctr. for Advanced Research in Biotechnology (United States)
Lori S. Goldner, National Institute of Standards and Technology (United States)


Published in SPIE Proceedings Vol. 5699:
Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III
Dan V. Nicolau; Ramesh Raghavachari; Dan V. Nicolau; Jörg Enderlein; Robert C. Leif; Daniel L. Farkas, Editor(s)

© SPIE. Terms of Use
Back to Top