MOLECULAR CHARACTERIZATION OF A RECOMBINANT POXVIRUS.
Item
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Title
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MOLECULAR CHARACTERIZATION OF A RECOMBINANT POXVIRUS.
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Identifier
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AAI8611326
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identifier
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8611326
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Creator
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BERKOWITZ, ELCHONON MICHAEL.
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Contributor
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Beatriz G.T. Pogo
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Date
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1986
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, Molecular
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Abstract
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Vaccinia virus is a cytolytic orthopoxvirus and Shope fibroma virus is a tumorigenic leporipoxvirus. SFV-I, a fibroma isolate, is tumorigenic in vivo, but unlike other leporipoxviruses, is cytolytic rather than focus-forming in vitro. Antibodies raised against either vaccinia virus or SFV neutralize the infectivity of SPV-I. These observations suggested that SFV-I might contain genetic information from both orthopox and leporipox viruses.;SFV-I DNA was digested with various restriction enzymes and analyzed by comparison to patterns generated by the restriction endonuclease digestion of vaccinia virus and SFV-W DNAs. SFV-I DNA showed great similarity to vaccinia virus DNA with all enzymes used, but not to SFV-W DNA.;The genetic homology between vaccinia virus and SFV-I DNAs was determined by the use of nick-translated probes to hybridize to restriction endonuclease digested DNA transferred to nitro-cellulose filters. These experiments showed that vaccinia virus and SFV-I were closely related; even fragments that migrated differently in agarose gels hybridized.;The genetic relatedness between the DNAs of orthopoxviruses, with vaccinia virus serving as the prototype, and leporipoxviruses, with SFV-W as the prototype, was also determined by Southern hybridizations. It was shown that orthopoxviruses and leporipoxviruses had no sequences in common.;When SFV-I DNA was probed with nick-translated SFV-W DNA, a subset of fragments were shown to cross-hybridize. This was also the case when the reverse experiment, in which SFV-I was the probe, was performed. These experiments showed that SFV-I, while an orthopox, also contained leporipoxvirus-related DNA.;Comparative studies of proteins of an orthopoxvirus, leporipoxvirus and SFV-I were also carried out. ('35)S-methionine-labeled virion proteins and immunoprecipitation of virion proteins by heterologous antisera did not prove useful in identifying proteins specific for orthopoxviruses. ('32)P-labeled phophoproteins were in some instances shared by all three viruses: in others, they were genus-specific. Glucosamine-containing proteins of orthopoxviruses and leporipoxviruses, however, were different.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
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Program
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Biomedical Sciences
