Work

Approaches for stoichiometry and distance determination of nanometer bio-complex by dual-channel single molecule imaging

Year: 2009

Type: conference-paper

Abstract: A dual-channel imaging system with single fluorophore sensitivity was assembled in this lab. Inclusion of an integrated laser combiner was introduced to facilitate simultaneous dual-channel imaging. The imaging system has been applied to study the structure, stoichiometry, distance and function of the phi29 DNA packaging motor. Approaches including single molecule photobleaching, single molecule FRET and binomial distribution quantification were carried out to clarify the stoichiometry and distance of pRNA on the biologically active packaging motor. The results were statistically analyzed to access the copy number of pRNA, and the distance constraint was used to verify the 3D structure of the computer model of phi29 DNA packaging motor.

Source: IEEE/NIH Life Science Systems and Applications Workshop

DOI: 10.1109/lissa.2009.4906725

Authors: Hui Zhang, Dan Shu, Mark Browne, Peixuan Guo

Institution: University of Cincinnati

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Eye Institute, National Institute of General Medical Sciences, National Institute of Biomedical Imaging and Bioengineering

Awards: G1533765926, G2413273428, Structure and Function of Phi29 hexameric pRNA