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Purification and elementary analysis of Streptomyces griseus phage

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Field Value
Title Purification and elementary analysis of Streptomyces griseus phage
Names South, Dorothy Jean (creator)
Gilmour, C. M. (advisor)
Date Issued 1966-05-13 (iso8601)
Note Graduation date: 1966
Abstract This investigation was undertaken to provide a chemical
description of an actinophage for Streptomyces griseus, and to
determine the base composition of the phage DNA moiety. It was
believed of interest to learn whether the actinophage for the
filamentous S. griseus host would be different in its chemical
properties from the bacteriophages. No such chemical analysis
of an actinophage has been reported. Complete chemical
analyses are available only for a few of the Escherichia coli
phages.
A method for isolation and purification of the phage was
developed, which consisted of isolation by several cycles of
differential centrifugation and final purification by centrifugation
in a cesium chloride density gradient. The purified phage was
analyzed for phosphorus, DNA, nitrogen and protein by means of standard analytical methods and the resulting data reported,
relative to the dry weight of the purified phage. The DNA was
extracted from purified phage by the phenol extraction procedure.
The purine and pyrimidine base composition of the DNA was
determined by standard methods for the hydrolysis of DNA and
analysis for the bases by paper chromatography, and also by
calculation from the thermal denaturation temperature, or
melting point, of the DNA.
It was concluded that the S. griseus phage under study
did not differ significantly in its chemical properties from most
of the other bacteriophages that have been similarly analyzed.
It was shown to consist of about 60% DNA and 40% protein. The
DNA content of 60% was somewhat higher than the 50% value
generally reported for phages. The analysis of the extracted
DNA for purine and pyrimidine bases showed it to be composed
of the four usual DNA bases. There was no evidence for the
presence of an unusual base, The DNA base composition value
of 53.4% guanine plus cytosine determined by chromatographic
analysis agrees within experimental error with the 51.4%
value calculated from the thermal denaturation temperature.
The buoyant density in cesium chloride was found to be 1.55 g/cc.
Genre Thesis/Dissertation
Topic Bacteriophages
Identifier http://hdl.handle.net/1957/49022

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