| fullrecord |
[{"key": "dc.contributor.author", "value": "Gr\u00f6nroos, Toni", "language": null, "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2012-05-16T09:42:11Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2012-05-16T09:42:11Z", "language": null, "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2012", "language": null, "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.other", "value": "oai:jykdok.linneanet.fi:1216783", "language": null, "element": "identifier", "qualifier": "other", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/37850", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Bradavidiini on Bradyrhizobium japonicum bakteerista l\u00f6ydetty avidiinin ja streptaviidiinin kaltainen proteiini. Avidiini on per\u00e4isin kanasta ja streptavidiini Streptomyces avidinii bakteerista. Bradavidiinin aminohapposekvenssi on noin 30 prosenttisesti samankaltainen avidiinin ja streptavidiinin ydinalueiden sekvenssien kanssa. Bradavidiini koostuu 138 aminohaposta ja C-terminaalisesti pienennetty bradavidiini, ns. bradcore, 118 aminohaposta. Bradavidiinilla on useimpien avidiinien tapaan tetrameerinen kvaternaarirakenne ja se muistuttaa sek\u00e4 rakenteeltaan ett\u00e4 toiminnaltaan avidiinia ja streptavidiinia. Bradavidiinilla on muiden avidiinien tapaan kyky sitoa voimakkaasti D-biotiinia. \nAvidiinien sitoutuminen biotiiniin on johtanut niiden laajaan k\u00e4ytt\u00f6\u00f6n avidiini-biotiini teknologiassa, esim. l\u00e4\u00e4ketieteess\u00e4 geeniterapiaan ja kuvantamiseen. Avidiini ja streptavidiini ovat my\u00f6s stabiileja l\u00e4mm\u00f6n ja pHn vaihteluille sek\u00e4 proteolyyttisille entsyymeille. Immunologisilta ominaisuuksiltaan bradavidiini on kuitenkin (strept)avidiineista poikkeava. J\u00e4niksen ja ihmisen streptavidiinia tunnistavat seerumit eiv\u00e4t reagoineet bradavidiinin kanssa merkitt\u00e4v\u00e4sti. Avidiinista ja streptavidiinista poiketen, t\u00e4ysimittaisen bradavidiinin karboksyylip\u00e4\u00e4ss\u00e4 on lyhyt aminohappoh\u00e4nt\u00e4, joka kykenee sitoutumaan viereisen monomeerin biotiinia sitovaan kohtaan ja mahdollisesti est\u00e4m\u00e4\u00e4n biotiinin sitoutumisen.\nStrep-tag ja Strep-tag II ovat yhdeks\u00e4n aminohappot\u00e4hteen mittaisia peptidej\u00e4, jotka sitoutuvat luontaisesti ja palautuvasti streptavidiinin biotiinia sitovaan kohtaan. Strep-tag ja Strep-tag II peptidej\u00e4 k\u00e4ytet\u00e4\u00e4n mm. affiniteettikahvoina rekombinanttiproteiinien puhdistuksessa sek\u00e4 analyyttisin\u00e4 ty\u00f6kaluina immunokemiallisissam\u00e4\u00e4rityksiss\u00e4. Strep-tag II kykenee Strep-tagista poiketen sitoutumaan streptavidiinin biotiinia sitovaan kohtaan sek\u00e4 C- ett\u00e4 N-terminaalisena fuusiona. Brad-tag on kahdeksan aminohappot\u00e4hteen pituinen peptidi, jonka sekvenssi (SEKLSNTK) l\u00f6ydettiin bradavidiinin karboksyylip\u00e4\u00e4ss\u00e4 olevasta aminohappoh\u00e4nn\u00e4st\u00e4. \nIsotermisella tirauskalorimetrilla (ITC) tehtyjen mittausten perusteella bradpeptidi sitoutuu bradavidiiniin, mutta ei sitoudu avidiiniin tai rhizavidiiniin. Bradpeptidin affiniteetti bradavidiinin C-terminaalisesti lyhennettyyn muotoon, bradcoreen, on hieman suurempi kuin Strep-tag ja Strep-tag II peptidien affiniteetti streptavidiiniin. N\u00e4iden lupaavien tutkimusten perusteella tuotettiin tehostetun vihre\u00e4 fluoresoivan proteiinin (EGFP) kanssa bradtag-EGFP fuusioproteiineja. Fuusio tehtiin joko EGFPn C- tai N-terminaaliin. Tarkoituksena oli selvitt\u00e4\u00e4 toimiiko Brad-tag fuusioproteiinissa ja onko C- tai N-terminaalisilla fuusioilla eroa. Mutatoitujen bradpeptidien affiniteettia bradcoreen pyrittiin selvitt\u00e4m\u00e4\u00e4n ITC-mittauksilla.\nBradtag-EGFP fuusioproteiineja saatiin tuotettua, kuitenkin N-terminaalisen Brad-tagin sis\u00e4lt\u00e4vien fuusioproteiinien tuottotasot olivat huomattavasti C-terminaalisia Brad-tag fuusioproteiineja korkeammat. Erilaisilla bradcore-resiiniin tehdyill\u00e4 sitoutus- ja puhdistuskokeilla selvitettiin, ett\u00e4 Brad-tag sitoutuu EGFPn aminoterminaalisena fuusiona bradcoren biotiinia sitovaan kohtaan ja sit\u00e4 voidaan k\u00e4ytt\u00e4\u00e4 aminoterminaalisena affiniteettikahvana. Mutaatioilla havaittiin olevan heikent\u00e4v\u00e4 vaikutus bradpeptidien ja bradcoren v\u00e4lisess\u00e4 sitoutumisessa.", "language": null, "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted using Plone Publishing form by Toni Gr\u00f6nroos (tohegron) on 2012-05-16 09:42:11.104783. Form: Pro gradu -lomake (1 tekij\u00e4) (https://kirjasto.jyu.fi/julkaisut/julkaisulomakkeet/pro-gradu-lomake-1-tekijae). JyX data:", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by jyx lomake-julkaisija (jyx-julkaisija@noreply.fi) on 2012-05-16T09:42:11Z\nNo. of bitstreams: 2\nURN:NBN:fi:jyu-201205161669.pdf: 1190530 bytes, checksum: 1383a26bc9760bc7787e95a9b6701344 (MD5)\nlicense.html: 5033 bytes, checksum: 34b03173fe0155c210bc176c9415952c (MD5)", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2012-05-16T09:42:11Z (GMT). No. of bitstreams: 2\nURN:NBN:fi:jyu-201205161669.pdf: 1190530 bytes, checksum: 1383a26bc9760bc7787e95a9b6701344 (MD5)\nlicense.html: 5033 bytes, checksum: 34b03173fe0155c210bc176c9415952c (MD5)\n Previous issue date: 2012", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "59 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": "fin", "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": "affiniteettikahva", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.title", "value": "Bradavidiinista johdetun bradpeptidin karakterisointi ja soveltuvuus affiniteettikromatografiaan", "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-201205161669", "language": null, "element": "identifier", "qualifier": "urn", "schema": "dc"}, {"key": "dc.type.dcmitype", "value": "Text", "language": "en", "element": "type", "qualifier": "dcmitype", "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": "2012-05-16T09:42:11Z", "language": null, "element": "date", "qualifier": "updated", "schema": "dc"}, {"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": "avidiini", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "streptavidiini", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "proteiinit", "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": "dc.type.okm", "value": "G2", "language": null, "element": "type", "qualifier": "okm", "schema": "dc"}]
|