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investigation of the nonstructural proteins of mammalian reovirus serotype 3: Mu NS and sigma NS

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Date Issued:
2002
Summary:
The reovirus nonstructural proteins muNS and sigmaNS are thought to play a role in assortment. To produce large quantities of the reovirus nonstructural proteins, two recombinant vaccinia virus systems were used, one for expression of muNS and the other for expression of muNS. The reovirus gene is under the control of the phage T7 RNA polymerase promoter. A recombinant vaccinia virus containing the gene for T7 RNA polymerase was added and protein expression was determined. Protein expression was confirmed by infecting mouse fibroblast L929 cells, harvesting the cells and running an SDS-PAGE gel, followed by western blot, followed by Western-immunoperoxidase assay using a polyclonal antibody for the reovirus proteins, or by radioimmunoprecipitation followed by an SDS-PAGE gel. Purification of the recombinant proteins was accomplished by ammonium sulfate fractionation and column chromatography. The purified proteins will be utilized to further investigate the role of the reovirus nonstructural proteins in assortment.
Title: An investigation of the nonstructural proteins of mammalian reovirus serotype 3: Mu NS and sigma NS.
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Name(s): Van Vliet, Kim Marie
Florida Atlantic University, Degree Grantor
Roner, Michael R., Thesis Advisor
Type of Resource: text
Genre: Electronic Thesis Or Dissertation
Issuance: monographic
Date Issued: 2002
Publisher: Florida Atlantic University
Place of Publication: Boca Raton, Fla.
Physical Form: application/pdf
Extent: 63 p.
Language(s): English
Summary: The reovirus nonstructural proteins muNS and sigmaNS are thought to play a role in assortment. To produce large quantities of the reovirus nonstructural proteins, two recombinant vaccinia virus systems were used, one for expression of muNS and the other for expression of muNS. The reovirus gene is under the control of the phage T7 RNA polymerase promoter. A recombinant vaccinia virus containing the gene for T7 RNA polymerase was added and protein expression was determined. Protein expression was confirmed by infecting mouse fibroblast L929 cells, harvesting the cells and running an SDS-PAGE gel, followed by western blot, followed by Western-immunoperoxidase assay using a polyclonal antibody for the reovirus proteins, or by radioimmunoprecipitation followed by an SDS-PAGE gel. Purification of the recombinant proteins was accomplished by ammonium sulfate fractionation and column chromatography. The purified proteins will be utilized to further investigate the role of the reovirus nonstructural proteins in assortment.
Identifier: 9780493772868 (isbn), 12930 (digitool), FADT12930 (IID), fau:9802 (fedora)
Note(s): Thesis (M.S.)--Florida Atlantic University, 2002.
Subject(s): Reoviruses
RNA viruses
Held by: Florida Atlantic University Libraries
Persistent Link to This Record: http://purl.flvc.org/fcla/dt/12930
Sublocation: Digital Library
Use and Reproduction: Copyright © is held by the author with permission granted to Florida Atlantic University to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
Use and Reproduction: http://rightsstatements.org/vocab/InC/1.0/
Host Institution: FAU
Is Part of Series: Florida Atlantic University Digital Library Collections.