Single-particle light microscopy of bacteriophages

Philip Serwer, Wang Houyi

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Bacteriophage capsids have a protein shell with a symmetrical, fullerene-like arrangement of sub-units. In the case of double-stranded DNA bacteriophages, the capsid joins with accessory proteins to form a DNA packaging motor that packages a genome in a cavity of the capsid. The motor cleaves ATP to obtain the needed energy. Light microscopy of single bacteriophages and single bacteriophage DNA packaging intermediates is being developed for the following reasons: (1) A synchronization-independent, fractionation-independent procedure is needed for the in vitro analysis of bacteriophage DNA packaging motors. (2) A non-biological procedure is needed for identifying and characterizing new bacteriophages needed for studies of bacteriophage gene homologies. In a recent study, light microscopy-based nanometry is used to follow the in vitro packaging of DNA in real time. Fluorescence microscopy of stained DNA is similarly used. Towards a more thorough analysis by fluorescence microscopy, single bacteriophage capsids are visualized by the unenhanced fluorescence of covalently bound protein-specific dyes. Dimerization of capsids is observed in real time. The dimerizing capsids had been restricted to a thin planar zone so that single-particle tracking was performed before, during, and after dimerization. Photobleaching is not a major problem. Thermal motion-based procedures are used for distinguishing binding from accidental co-migration. The long-range objective is the simultaneous real time monitoring of multiple state variables during cycling of a single DNA packaging motor. The results of these basic studies are applicable to both nanotechnological drug delivery and biological therapy.

Original languageEnglish (US)
Pages (from-to)2014-2028
Number of pages15
JournalJournal of Nanoscience and Nanotechnology
Volume5
Issue number12
DOIs
StatePublished - Dec 2005

Fingerprint

Bacteriophages
Optical microscopy
DNA Packaging
Microscopy
Capsid
DNA
Light
Packaging
Dimerization
Fluorescence microscopy
Proteins
Fluorescence Microscopy
Genes
Fullerenes
Photobleaching
Biological Therapy
Adenosinetriphosphate
Accessories
Fractionation
Drug delivery

Keywords

  • Bacteriophages φ29, λ, and t7
  • Fluorescence Microscopy
  • Fluorescent Dyes
  • In vitro procedures
  • Single-nanoparticle biochemistry

ASJC Scopus subject areas

  • Chemistry(all)
  • Materials Science(all)
  • Materials Science (miscellaneous)
  • Engineering (miscellaneous)

Cite this

Single-particle light microscopy of bacteriophages. / Serwer, Philip; Houyi, Wang.

In: Journal of Nanoscience and Nanotechnology, Vol. 5, No. 12, 12.2005, p. 2014-2028.

Research output: Contribution to journalArticle

@article{867ec7746b954be69125da6544fd124b,
title = "Single-particle light microscopy of bacteriophages",
abstract = "Bacteriophage capsids have a protein shell with a symmetrical, fullerene-like arrangement of sub-units. In the case of double-stranded DNA bacteriophages, the capsid joins with accessory proteins to form a DNA packaging motor that packages a genome in a cavity of the capsid. The motor cleaves ATP to obtain the needed energy. Light microscopy of single bacteriophages and single bacteriophage DNA packaging intermediates is being developed for the following reasons: (1) A synchronization-independent, fractionation-independent procedure is needed for the in vitro analysis of bacteriophage DNA packaging motors. (2) A non-biological procedure is needed for identifying and characterizing new bacteriophages needed for studies of bacteriophage gene homologies. In a recent study, light microscopy-based nanometry is used to follow the in vitro packaging of DNA in real time. Fluorescence microscopy of stained DNA is similarly used. Towards a more thorough analysis by fluorescence microscopy, single bacteriophage capsids are visualized by the unenhanced fluorescence of covalently bound protein-specific dyes. Dimerization of capsids is observed in real time. The dimerizing capsids had been restricted to a thin planar zone so that single-particle tracking was performed before, during, and after dimerization. Photobleaching is not a major problem. Thermal motion-based procedures are used for distinguishing binding from accidental co-migration. The long-range objective is the simultaneous real time monitoring of multiple state variables during cycling of a single DNA packaging motor. The results of these basic studies are applicable to both nanotechnological drug delivery and biological therapy.",
keywords = "Bacteriophages φ29, λ, and t7, Fluorescence Microscopy, Fluorescent Dyes, In vitro procedures, Single-nanoparticle biochemistry",
author = "Philip Serwer and Wang Houyi",
year = "2005",
month = "12",
doi = "10.1166/jnn.2005.447",
language = "English (US)",
volume = "5",
pages = "2014--2028",
journal = "Journal of Nanoscience and Nanotechnology",
issn = "1533-4880",
publisher = "American Scientific Publishers",
number = "12",

}

TY - JOUR

T1 - Single-particle light microscopy of bacteriophages

AU - Serwer, Philip

AU - Houyi, Wang

PY - 2005/12

Y1 - 2005/12

N2 - Bacteriophage capsids have a protein shell with a symmetrical, fullerene-like arrangement of sub-units. In the case of double-stranded DNA bacteriophages, the capsid joins with accessory proteins to form a DNA packaging motor that packages a genome in a cavity of the capsid. The motor cleaves ATP to obtain the needed energy. Light microscopy of single bacteriophages and single bacteriophage DNA packaging intermediates is being developed for the following reasons: (1) A synchronization-independent, fractionation-independent procedure is needed for the in vitro analysis of bacteriophage DNA packaging motors. (2) A non-biological procedure is needed for identifying and characterizing new bacteriophages needed for studies of bacteriophage gene homologies. In a recent study, light microscopy-based nanometry is used to follow the in vitro packaging of DNA in real time. Fluorescence microscopy of stained DNA is similarly used. Towards a more thorough analysis by fluorescence microscopy, single bacteriophage capsids are visualized by the unenhanced fluorescence of covalently bound protein-specific dyes. Dimerization of capsids is observed in real time. The dimerizing capsids had been restricted to a thin planar zone so that single-particle tracking was performed before, during, and after dimerization. Photobleaching is not a major problem. Thermal motion-based procedures are used for distinguishing binding from accidental co-migration. The long-range objective is the simultaneous real time monitoring of multiple state variables during cycling of a single DNA packaging motor. The results of these basic studies are applicable to both nanotechnological drug delivery and biological therapy.

AB - Bacteriophage capsids have a protein shell with a symmetrical, fullerene-like arrangement of sub-units. In the case of double-stranded DNA bacteriophages, the capsid joins with accessory proteins to form a DNA packaging motor that packages a genome in a cavity of the capsid. The motor cleaves ATP to obtain the needed energy. Light microscopy of single bacteriophages and single bacteriophage DNA packaging intermediates is being developed for the following reasons: (1) A synchronization-independent, fractionation-independent procedure is needed for the in vitro analysis of bacteriophage DNA packaging motors. (2) A non-biological procedure is needed for identifying and characterizing new bacteriophages needed for studies of bacteriophage gene homologies. In a recent study, light microscopy-based nanometry is used to follow the in vitro packaging of DNA in real time. Fluorescence microscopy of stained DNA is similarly used. Towards a more thorough analysis by fluorescence microscopy, single bacteriophage capsids are visualized by the unenhanced fluorescence of covalently bound protein-specific dyes. Dimerization of capsids is observed in real time. The dimerizing capsids had been restricted to a thin planar zone so that single-particle tracking was performed before, during, and after dimerization. Photobleaching is not a major problem. Thermal motion-based procedures are used for distinguishing binding from accidental co-migration. The long-range objective is the simultaneous real time monitoring of multiple state variables during cycling of a single DNA packaging motor. The results of these basic studies are applicable to both nanotechnological drug delivery and biological therapy.

KW - Bacteriophages φ29, λ, and t7

KW - Fluorescence Microscopy

KW - Fluorescent Dyes

KW - In vitro procedures

KW - Single-nanoparticle biochemistry

UR - http://www.scopus.com/inward/record.url?scp=33645828216&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33645828216&partnerID=8YFLogxK

U2 - 10.1166/jnn.2005.447

DO - 10.1166/jnn.2005.447

M3 - Article

C2 - 16430135

AN - SCOPUS:33645828216

VL - 5

SP - 2014

EP - 2028

JO - Journal of Nanoscience and Nanotechnology

JF - Journal of Nanoscience and Nanotechnology

SN - 1533-4880

IS - 12

ER -