An investigation into the specific function of the vaccinia virus 13.8 kDa protein encoded by the N1

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dc.contributor.advisorKotwal, Girish Jen_ZA
dc.contributor.authorAbrahams, Melissa-Rose Hildaen_ZA
dc.date.accessioned2014-10-20T07:38:17Z
dc.date.available2014-10-20T07:38:17Z
dc.date.issued2005en_ZA
dc.descriptionIncludes bibliographical references.en_ZA
dc.description.abstractVaccinia virus is the most extensively studied, prototype vertebrate poxvirus, which was used as a vaccine in the eradication of smallpox. The genome of this virus has characteristic variable termini encoding open reading frames that are not essential for virus replication in cell culture. One such open reading frame, N1L situated at the left terminal region of the neurovirulent Western Reserve (WR) vaccinia virus strain, encodes a protein 13.8 kDa in size. In vivo studies in mouse brains revealed that a recombinant virus, vGK5, tacking the expression of the 13.8 kDa protein was rendered replication deficient in the brain. An essential requirement of poxviruses for their replication is the energy molecule adenosine triphosphate (ATP). The supply of this molecule in the brain to support replication of a virus is limited due to the high-energy requirements and small energy reserves of this organ. The specific function of the vaccinia virus 13.8 kDa protein in relation to viral replication in the brain was investigated. The South African (SA) Lister vaccinia virus strain was confirmed to encode an identical N1L gene to that of the WR vaccinia virus by amplification, cloning and sequencing of the Lister N1L open reading frame. The Lister vaccinia virus and a 13.8 kDa deletion strain (vGK5) were cultivated and used to intracranially infect mice. Using a luciferin/luciferase bioluminescence assay system the ATP levels in Lister and vGK5 vaccinia virus-infected mouse brains were measured and found to differ significantly after a 5-day infection period. The SA vaccine Lister vaccinia virus strain was found to be a slow growing virus in the brain. Subsequently, a possible role for the vaccinia virus 13.8 kDa protein in influencing ATP levels in the brain was postulated, yet a neurovirulent wild type strain is needed for further studies to consolidate this result. The 13.8 kDa protein was successfully expressed in the P. pastoris yeast expression system and positively identified by immunodetection studies.en_ZA
dc.identifier.apacitationAbrahams, M. H. (2005). <i>An investigation into the specific function of the vaccinia virus 13.8 kDa protein encoded by the N1</i>. (Thesis). University of Cape Town ,Faculty of Health Sciences ,Division of Virology. Retrieved from http://hdl.handle.net/11427/8637en_ZA
dc.identifier.chicagocitationAbrahams, Melissa-Rose Hilda. <i>"An investigation into the specific function of the vaccinia virus 13.8 kDa protein encoded by the N1."</i> Thesis., University of Cape Town ,Faculty of Health Sciences ,Division of Virology, 2005. http://hdl.handle.net/11427/8637en_ZA
dc.identifier.citationAbrahams, M. 2005. An investigation into the specific function of the vaccinia virus 13.8 kDa protein encoded by the N1. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Abrahams, Melissa-Rose Hilda AB - Vaccinia virus is the most extensively studied, prototype vertebrate poxvirus, which was used as a vaccine in the eradication of smallpox. The genome of this virus has characteristic variable termini encoding open reading frames that are not essential for virus replication in cell culture. One such open reading frame, N1L situated at the left terminal region of the neurovirulent Western Reserve (WR) vaccinia virus strain, encodes a protein 13.8 kDa in size. In vivo studies in mouse brains revealed that a recombinant virus, vGK5, tacking the expression of the 13.8 kDa protein was rendered replication deficient in the brain. An essential requirement of poxviruses for their replication is the energy molecule adenosine triphosphate (ATP). The supply of this molecule in the brain to support replication of a virus is limited due to the high-energy requirements and small energy reserves of this organ. The specific function of the vaccinia virus 13.8 kDa protein in relation to viral replication in the brain was investigated. The South African (SA) Lister vaccinia virus strain was confirmed to encode an identical N1L gene to that of the WR vaccinia virus by amplification, cloning and sequencing of the Lister N1L open reading frame. The Lister vaccinia virus and a 13.8 kDa deletion strain (vGK5) were cultivated and used to intracranially infect mice. Using a luciferin/luciferase bioluminescence assay system the ATP levels in Lister and vGK5 vaccinia virus-infected mouse brains were measured and found to differ significantly after a 5-day infection period. The SA vaccine Lister vaccinia virus strain was found to be a slow growing virus in the brain. Subsequently, a possible role for the vaccinia virus 13.8 kDa protein in influencing ATP levels in the brain was postulated, yet a neurovirulent wild type strain is needed for further studies to consolidate this result. The 13.8 kDa protein was successfully expressed in the P. pastoris yeast expression system and positively identified by immunodetection studies. DA - 2005 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 2005 T1 - An investigation into the specific function of the vaccinia virus 13.8 kDa protein encoded by the N1 TI - An investigation into the specific function of the vaccinia virus 13.8 kDa protein encoded by the N1 UR - http://hdl.handle.net/11427/8637 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/8637
dc.identifier.vancouvercitationAbrahams MH. An investigation into the specific function of the vaccinia virus 13.8 kDa protein encoded by the N1. [Thesis]. University of Cape Town ,Faculty of Health Sciences ,Division of Virology, 2005 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/8637en_ZA
dc.language.isoengen_ZA
dc.publisher.departmentDivision of Virologyen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.subject.otherMedical Virologyen_ZA
dc.titleAn investigation into the specific function of the vaccinia virus 13.8 kDa protein encoded by the N1en_ZA
dc.typeMaster Thesis
dc.type.qualificationlevelMasters
dc.type.qualificationnameMScen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceThesisen_ZA
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