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[{"key": "dc.contributor.advisor", "value": "Kalliolinna, Olli", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Vihinen-Ranta, Maija", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Kleemola, Ari", "language": null, "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2015-05-06T21:24:44Z", "language": "", "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2015-05-06T21:24:44Z", "language": "", "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2015", "language": null, "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.other", "value": "oai:jykdok.linneanet.fi:1473259", "language": null, "element": "identifier", "qualifier": "other", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/45796", "language": "", "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Koiran parvovirus (CPV) on halkaisijaltaan 26 nm vaipaton virus. Ikosahedraalinen kapsidi sis\u00e4lt\u00e4\u00e4 yksijuosteisen ~5kb DNA-genomin. CPV sis\u00e4lleotetaan soluun endosytoosilla, jonka j\u00e4lkeen se kulkeutuu\r\nendosomin sis\u00e4ll\u00e4 kohti tumaa. CPV vapautuu solulimaan tumaa ymp\u00e4r\u00f6ivist\u00e4 lysosomeista. Genomin replikaatio ja uusien virionien kasautuminen tapahtuvat tumassa. Aikaisemmin on oletettu, ett\u00e4 parvovirukset\r\ntodenn\u00e4k\u00f6isesti p\u00e4\u00e4sisiv\u00e4t tumaan tumahuokosen (NPC) kautta. Uudet tutkimukset kuitenkin viittaavat siihen ett\u00e4 siirtyminen tumaan voisi tapahtua toista reitti\u00e4 k\u00e4ytt\u00e4en.\r\n\r\nCPV ja NPC proteiinien (Nup) kolokalisaatio-kokeissa selvitettiin voidaanko tiettyj\u00e4 vuorovaikutuksia\r\nhavaita ja miss\u00e4 mahdolliset vuorovaikutukset sijaitsevat. N\u00e4iss\u00e4 kokeissa k\u00e4ytettiin uutta in situ proximity ligation assay (PLA) -menetelm\u00e4\u00e4, joka tuottaa fluoresenssi-signaalin kahteen eri kohdeproteiineihin sitoutuneiden vasta-aineiden sijaitessa l\u00e4hell\u00e4 toisiaan. Bromouridiini (BRU) on uridiinin analogi, joka voidaan tunnistaa spesifisti vasta-aineilla. Soluja viljeltiin BRU-mediumissa, jolloin BRU saatiin liitetty\u00e4 kehittyviin RNA-molekyyleihin. T\u00e4m\u00e4n j\u00e4lkeen oli mahdollista arvioida infektiosta johtuvia muutoksia\r\nRNA-synteesiss\u00e4. Fluoresenssia mitattiin virtaussytometrian ja konfokaalimikroskopian avulla.\r\n\r\nErityist\u00e4 kolokalisaatiota ei havaittu tumakalvolla Nup358 ja viruskapsidin v\u00e4lill\u00e4. Sen sijaan signaali esiintyi levinneen\u00e4 sytoplasmassa ja n\u00e4ytti siirtyv\u00e4n kohti tumaa infektion edetess\u00e4. Samankaltaista sytoplasmista kolokalisaatiota n\u00e4htiin infektoimattomissa soluissa Nup153 ja Nup358 vasta-aineilla suoritetuissa kokeissa.\r\nT\u00e4m\u00e4 signaali oli kuitenkin joissain soluissa vahvimmillaan tuman v\u00e4litt\u00f6m\u00e4ss\u00e4 l\u00e4heisyydess\u00e4. Infektion my\u00f6h\u00e4isess\u00e4 vaiheessa sytoplasmassa oli havaittavissa vahva signaali CPV kapsidi ja NS1 proteiinien v\u00e4lill\u00e4.\r\nImmunoleimaus ja virtaussytometri mittaukset osoittavat RNA-synteesin vaimentumista infektion seurauksena.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Canine parvovirus (CPV) is a non-enveloped virus with a 26 nm diameter icosahedral capsid. The capsid holds a ~5kb single-stranded DNA genome. CPV is internalized by endocytosis followed by intracellular trafficking inside an endosome towards the nucleus. CPV is released from perinuclear lysosomes followed by replication of the genome and assembly of progeny virions inside the nucleus. It has been previously assumed that parvoviruses are likely to enter the nucleus through the nuclear pore complex (NPC). Recent studies however suggest that entry to the nucleus might take place by an NPC-independent mechanism.\r\n\r\nColocalization of CPV and NPC proteins (Nups) were examined to find out if specific interactions could be detected and if so, where these interactions would occur. Experiments were carried out by a novel in situ\r\nproximity ligation assay (PLA) method that generates fluorescence only when two antibodies bound to different target proteins are situated at close proximity. Bromouridine (BRU) is an uridine analog that can be\r\nspecifically bound by antibodies. Cells were cultured in the presence of BRU that became incorporated into nascent RNA molecules. It was then possible to estimate changes in RNA synthesis as a result of infection.\r\nThe fluorescence was measured by flow cytometry and visualized by confocal microscopy.\r\n\r\nColocalization at the nuclear envelope was not detected between Nup358 and CPV capsid. Instead, the signal was spread across the cytoplasm and appeared to be shifting towards the nucleus in time-dependent manner.\r\nCytoplasmic colocalization, sometimes located close to perinuclear region, was detected between Nup153 and Nup358 in noninfected cells. In a late phase of infection a very strong colocalization signal was seen\r\nbetween CPV capsid and NS1 proteins. Immunolabeling and flow cytometry assays suggest a decrease in the rate of RNA synthesis as a result of infection.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted using Plone Publishing form by Ari Kleemola (armaklee) on 2015-05-06 21:24:42.243090. Form: Pro gradu -lomake (https://kirjasto.jyu.fi/julkaisut/julkaisulomakkeet/pro-gradu-lomake). JyX data: [jyx_publishing-allowed (fi) =True]", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by jyx lomake-julkaisija (jyx-julkaisija@noreply.fi) on 2015-05-06T21:24:44Z\r\nNo. of bitstreams: 2\r\nURN:NBN:fi:jyu-201505071751.pdf: 14131619 bytes, checksum: b095384d5d6542826585e662111ed653 (MD5)\r\nlicense.html: 4869 bytes, checksum: 1b6af02385f87633cba02557536f1d00 (MD5)", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2015-05-06T21:24:44Z (GMT). No. of bitstreams: 2\r\nURN:NBN:fi:jyu-201505071751.pdf: 14131619 bytes, checksum: b095384d5d6542826585e662111ed653 (MD5)\r\nlicense.html: 4869 bytes, checksum: 1b6af02385f87633cba02557536f1d00 (MD5)\r\n Previous issue date: 2015", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "1 verkkoaineisto (44 sivua)", "language": null, "element": "format", "qualifier": "extent", "schema": "dc"}, {"key": "dc.format.mimetype", "value": "application/pdf", "language": null, "element": "format", "qualifier": "mimetype", "schema": "dc"}, {"key": "dc.language.iso", "value": "eng", "language": null, "element": "language", "qualifier": "iso", "schema": "dc"}, {"key": "dc.rights", "value": "In Copyright", "language": "en", "element": "rights", "qualifier": null, "schema": "dc"}, {"key": "dc.subject.other", "value": "CPV", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "PLA", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "NPC", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "Nup", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.title", "value": "Intracellular interplay between canine parvovirus with nuclear pore complex and effect of infection on RNA synthesis", "language": null, "element": "title", "qualifier": null, "schema": "dc"}, {"key": "dc.type", "value": "master thesis", "language": null, "element": "type", "qualifier": null, "schema": "dc"}, {"key": "dc.identifier.urn", "value": "URN:NBN:fi:jyu-201505071751", "language": null, "element": "identifier", "qualifier": "urn", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Pro gradu -tutkielma", "language": "fi", "element": "type", "qualifier": "ontasot", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Master\u2019s thesis", "language": "en", "element": "type", "qualifier": "ontasot", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Matemaattis-luonnontieteellinen tiedekunta", "language": "fi", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Faculty of Sciences", "language": "en", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.department", "value": "Bio- ja ymp\u00e4rist\u00f6tieteiden laitos", "language": "fi", "element": "contributor", "qualifier": "department", "schema": "dc"}, {"key": "dc.contributor.department", "value": "Department of Biological and Environmental Science", "language": "en", "element": "contributor", "qualifier": "department", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "University of Jyv\u00e4skyl\u00e4", "language": "en", "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "Jyv\u00e4skyl\u00e4n yliopisto", "language": "fi", "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Solu- ja molekyylibiologia", "language": "fi", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Cell and molecular biology", "language": "en", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "dc.date.updated", "value": "2015-05-06T21:24:45Z", "language": "", "element": "date", "qualifier": "updated", "schema": "dc"}, {"key": "yvv.contractresearch.funding", "value": "0", "language": "", "element": "contractresearch", "qualifier": "funding", "schema": "yvv"}, {"key": "dc.type.coar", "value": "http://purl.org/coar/resource_type/c_bdcc", "language": null, "element": "type", "qualifier": "coar", "schema": "dc"}, {"key": "dc.rights.accesslevel", "value": "openAccess", "language": "fi", "element": "rights", "qualifier": "accesslevel", "schema": "dc"}, {"key": "dc.type.publication", "value": "masterThesis", "language": null, "element": "type", "qualifier": "publication", "schema": "dc"}, {"key": "dc.subject.oppiainekoodi", "value": "4013", "language": null, "element": "subject", "qualifier": "oppiainekoodi", "schema": "dc"}, {"key": "dc.subject.yso", "value": "parvovirukset", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "tuma", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "virukset", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.format.content", "value": "fulltext", "language": null, "element": "format", "qualifier": "content", "schema": "dc"}, {"key": "dc.rights.url", "value": "https://rightsstatements.org/page/InC/1.0/", "language": null, "element": "rights", "qualifier": "url", "schema": "dc"}, {"key": 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