Concentration and purification of vesicular stomatitis virus by polyethylene glycol “precipitation”

McSharry, James J.; Benzinger, Rolf

doi:10.1016/0042-6822(70)90219-9

Cited by 99 publications

(40 citation statements)

References 7 publications

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“…Virus was precipitated from the culture medium with polyethylene glycol (McSharry & Benzinger, 1970). The precipitate was resuspended in TNE buffer (10 mM-Tris-HC1, 1 mM-EDTA, 150 mM-NaC1, pH 8.2) and centrifuged for 3 h at 90000g through a 25~ sucrose cushion.…”

Section: Methodsmentioning

confidence: 99%

Phenotypes of St Louis Encephalitis Virus Mutants Produced in Persistently Infected Mosquito Cell Cultures

Randolph

Hardy²

1988

Journal of General Virology

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SUMMARYViral mutants that appeared during long-term persistent infections of mosquito cell cultures (Aedes albopictus, A. dorsalis and Culex tarsalis) with St Louis encephalitis virus were characterized. Evidence was obtained for the presence of temperaturesensitive mutants in the A. dorsalis and C. tarsalis persistently infected cultures, and small plaque mutants were predominant in all cultures except one of two cell cultures of C. tarsalis. Virus from persistently infected A. albopictus ceU cultures was growthrestricted in Vero and C. tarsalis cells. One of two persistently infected A. dorsalis cell cultures also produced viral mutants that were growth-restricted in C. tarsalis cells. Further, Western blots of persistently infected A. albopictus cell extracts showed an overproduction of capsid (C) and envelope (E) structural proteins and reduced production of an Mr 27K protein (p27) which was immunologically related to the E protein. In contrast, the production of E and C proteins in persistently infected C. tarsalis cultures was consistent with the amount of infectious virus present, whereas p27 was relatively overproduced. These observations suggest that the host cell has an important influence on both the types and relative quantities of viral mutants that accumulate during long-term persistent infections.

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Section: Methodsmentioning

confidence: 99%

Phenotypes of St Louis Encephalitis Virus Mutants Produced in Persistently Infected Mosquito Cell Cultures

Randolph

Hardy²

1988

Journal of General Virology

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“…Con.-centration of the viruses was performed according to the method described by McSharry and Benzinger [14]. In brief, infected culture fluids were pooled and clarified by centrifugation at 6000 rpm for 30 min, from which viruses were precipitated with 6% polyethylene glycol 6000 (PEG) and 2.3% NaC1 at 4 C overnight.…”

Section: Methodsmentioning

confidence: 99%

Phenotypic Mixing of Vesicular Stomatitis Virus with HVJ (Sendai Virus)

Kimura¹

1973

Japanese Journal of Microbiology

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Vesicular stomatitis virus (VSV) plaque-forming virions obtained from mixed infection of HeLa cells with VSV and HVJ (Sendai virus) were shown to contain two types of phenotypically mixed virions : virions doubly neutralizable with both anti-VSV and anti-HVJ antisera (up to 15%), and virions which could be neutralized only with anti-HVJ serum and not with anti-VSV serum (up to 0.3%). No evidence for genetic recombination was obtained between these two viruses. Phenotypically mixed virions could adsorb to and elute from chiken erythrocytes, and bullet-shaped virions, typical of VSV, were found to be adsorbed to chicken erythrocytes in electron microscopy. Neuraminidase activity was detected in the virions specifically sedimented with anti-VSV serum. From these findings, phenotypically mixed virions were suggested to contain hemagglutinin and neuraminidase of HVJ in the envelope. In sucrose gradient centrifugation, phenotypically mixed virions enclosed within HVJ antigens sedimented slightly faster than standard HVJ.Simultaneous multiplication of two different viruses in cells can cause appearance of a progeny with varying characteristics. The occurrence of phenotypic mixing within the same group of viruses, such as myxoviruses [5,8] This paper describes evidence for the occurrence of phenotypic mixing between morphologically distinct groups of enveloped viruses, HVJ (Sendai virus) and VSV. Since HeLa cells produce only a few amount of HVJ progeny in a non-infectious form after a relatively long period of incubation, which is very expedient for investigation of phenotypic mixing, experiments have been performed mainly to clarify the characteristics of the VSV plaque-forming virions from mixed infection.

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“…Buffers were at 0.01 M and sodium acetate was used for the lowest pH and Tris HC1 buffer for the higher pH solutions. DISCUSSION Precipitation by PEG is of general utility as a method of cencentrating a host of biological macrostructures: bacteriophage, plant and animal viruses, ribosomes, DNA, and bacterial cells (7,8,9,10). These studies show the minimally required amount of PEG necessary for precipitation decreases for very asymmetric or large structures.…”

Section: Effect Of Phmentioning

confidence: 99%

Size fractionation of double-stranded DNA by precipitation with polyethylene glycol

1975

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We show that DNA molecules of differing molecular mass are separable by selective precipitation with polyethylene glycol (PEGt). Higher molecular mass DNA precipitates at lower PEG concentrations than lower molecular mass DNA. pouble-stranded DNA can be fractionated at least in the range of 3 X 10 to 1 X 107 daltons. The effects of PEG concentration, sodium chloride concentration, DNA concentration, pH, divalent ions, precipitation time, and centrifugal force have been determined. These studies show PEG precipitation offers a size fractionation method for DNA which is convenient, of high capacity, and applicable over a wide range of conditions. However, resolution is not high and separation of two species approaches 100% only if they differ in molecular mass by at least a factor of two.

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Concentration and purification of vesicular stomatitis virus by polyethylene glycol “precipitation”

Cited by 99 publications

References 7 publications

Phenotypes of St Louis Encephalitis Virus Mutants Produced in Persistently Infected Mosquito Cell Cultures

Phenotypes of St Louis Encephalitis Virus Mutants Produced in Persistently Infected Mosquito Cell Cultures

Phenotypic Mixing of Vesicular Stomatitis Virus with HVJ (Sendai Virus)

Size fractionation of double-stranded DNA by precipitation with polyethylene glycol

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