TY - JOUR
T1 - Agarose gel electrophoresis of bacteriophages and related particles
AU - Serwer, Philip
N1 - Funding Information:
For performing the experiment of Fig. 1, I thank Gary A. Griess and Shirley J. Hayes . For secretarial assistance, I thank Charlotte Farrell . Work in the author's laboratory was supported by grants from the National Institutes of Health (Grants A122568 and GM24365) and the Robert A . Welch Foundation (Grant AQ-764) .
PY - 1987/7/17
Y1 - 1987/7/17
N2 - Viruses and related particles have been fractionated by electrophoresis through gels. For agarose gels, the radius at the exclusion limit for spheres varies from 1500 nm in a 0.04% gel to 3.6 nm in a 4.0% gel. Thus, the size of the gel's pores can be adjusted to sieve all known viruses. By measurement of electrophoretic mobility (μ) as a function of agarose concentration, the μ in the absence of a solid support (μ0) can be determined for any particle. From the shape of a semilogarithmic plot of μ as a function of agarose percentage, a rod-shaped particle can be discriminated from a spherical particle. The sphere's radius can be determined from this plot with an accuracy of ±8%. Accuracy of ±1% has been more recently achieved using two-dimensional agarose gel electrophoresis. Though bacteriophages have been the primary object of study, the above techniques of agarose gel electrophoresis have also been applied to plant viruses and should be applicable to animal viruses. The μ0 values measured for bacteriophages with and without their tail fibers suggest a mechanism of controlling attachment to a host. A related mechanism is proposed for the control of the virulence of animal viruses. Measurement of outer radius for different forms of the capsid of bacteriophage P22 reveals variability in outer radius too small to be detected by electron microscopy.
AB - Viruses and related particles have been fractionated by electrophoresis through gels. For agarose gels, the radius at the exclusion limit for spheres varies from 1500 nm in a 0.04% gel to 3.6 nm in a 4.0% gel. Thus, the size of the gel's pores can be adjusted to sieve all known viruses. By measurement of electrophoretic mobility (μ) as a function of agarose concentration, the μ in the absence of a solid support (μ0) can be determined for any particle. From the shape of a semilogarithmic plot of μ as a function of agarose percentage, a rod-shaped particle can be discriminated from a spherical particle. The sphere's radius can be determined from this plot with an accuracy of ±8%. Accuracy of ±1% has been more recently achieved using two-dimensional agarose gel electrophoresis. Though bacteriophages have been the primary object of study, the above techniques of agarose gel electrophoresis have also been applied to plant viruses and should be applicable to animal viruses. The μ0 values measured for bacteriophages with and without their tail fibers suggest a mechanism of controlling attachment to a host. A related mechanism is proposed for the control of the virulence of animal viruses. Measurement of outer radius for different forms of the capsid of bacteriophage P22 reveals variability in outer radius too small to be detected by electron microscopy.
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U2 - 10.1016/0378-4347(87)80014-2
DO - 10.1016/0378-4347(87)80014-2
M3 - Review article
C2 - 3305547
AN - SCOPUS:0023658361
SN - 0378-4347
VL - 418
SP - 345
EP - 357
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
IS - C
ER -