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[{"key": "dc.contributor.advisor", "value": "Penttinen, Reetta", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Jalasvuori, Matti", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Norvasuo, Krista", "language": "", "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2022-06-21T08:47:33Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2022-06-21T08:47:33Z", "language": null, "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2022", "language": "", "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/81919", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Antibioottiresistentit bakteerit ovat yksi t\u00e4m\u00e4n p\u00e4iv\u00e4n suurimmista haasteista niiden horjuttaessa nykyaikaista terveydenhoitoa, ja ongelman ratkaisemiseksi tarvitaankin uudenlaisia l\u00e4hestymistapoja. Er\u00e4s potentiaalinen vaihtoehto on CRISPR-Cas9-systeemiin pohjautuvat geneettiset ty\u00f6kalut, jotka voidaan ohjelmoida katkaisemaan antibioottiresistenssin tuottava geeni, ja n\u00e4in ollen poistaa bakteerin vastustuskyky antibiootille. Bakteerisoluille soveltuvia yksisolumenetelmi\u00e4 on kehitetty bakteeriyhteis\u00f6jen tutkimiseksi yhden solun tarkkuudella. N\u00e4m\u00e4 menetelm\u00e4t mahdollistavat tietyn geenin is\u00e4nt\u00e4bakteerien tunnistamisen fuusio-PCR:n avulla, jossa taksonominen markkerigeeni (16S rRNA) liitet\u00e4\u00e4n tutkittavaan geeniin. T\u00e4ss\u00e4 ty\u00f6ss\u00e4 k\u00e4ytettiin yksisolu-ddPCR-menetelm\u00e4\u00e4 (Droplet Digital PCR) konjugaation avulla liikkuvan CRISPR-Cas9-plasmidisysteemin (cas9) ja blaCTX-M-15 resistenssigeenin kantajien tunnistamiseen bakteeripopulaatiossa. Aluksi kartoitettiin fuusio-PCR-reaktiolle suotuisat olosuhteet, jonka j\u00e4lkeen blaCTX-M-15 ja cas9 -geenien kantajat sek\u00e4 mahdollinen liikkuminen j\u00e4ljitettiin yksinkertaisessa synteettisess\u00e4 bakteeripopulaatiossa. Tulosten perusteella molempien geenien kantajat pystyttiin tunnistamaan yhteis\u00f6st\u00e4 yksisolumenetelm\u00e4n avulla. Havaittiin my\u00f6s, ett\u00e4 \nCRISRP-Cas9-plasmidisysteemi\u00e4 kantava Escherichia coli ei menestynyt hyvin, mutta bakteerien p\u00e4ivitt\u00e4inen migraatio populaatioon auttoi pit\u00e4m\u00e4\u00e4n sen osana yhteis\u00f6\u00e4. Tutkimustulosten avulla saatiin tietoa CRISPR-antimikrobiaalien tehokkuudesta heterogeenisess\u00e4 bakteeriyhteis\u00f6ss\u00e4, jossa niiden toimintakykyyn vaikuttavat monitahoiset interaktiot.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Bacterial resistance is one of the biggest threats in the modern healthcare. To resolve this problem, new approaches are needed. One potential solution is genetic engineering tools based on CRISPR-Cas9 system, that can be directed to cut target antibiotic resistance genes, reducing antibiotic resistance. Recently, microbial single-cell methods have been developed for analyzing bacterial communities at single cell resolution. By utilizing fusion PCR reaction for combining taxonomic\n16S rRNA marker gene with the gene of interest, the bacteria harbouring the target gene can be identified. In this study, single-cell method based on ddPCR (Droplet Digital PCR) was used to trace the dispersal of conjugative CRISPR-Cas9 plasmid system (cas9), and blaCTX-M-15 resistance gene in heterogenous bacterial population. First, optimized conditions for fusion PCR reaction were determined. Carriers of cas9 and blaCTX-M-15 genes, and the potential transfer within bacteria were detected in synthetic bacterial population. The results show that using\nsingle-cell method the carriers of these genes were identified in bacterial community. It was observed that in 7-day co-culture experiment, Escherichia coli delivery strain of CRISPR-Cas9 plasmid system did not survive, but daily migration into the population helped to maintain it as part of the community. These results provide knowledge of the efficiency of CRISPR-antimicrobials in heterogenous bacterial community, where complex interactions may affect their competence.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by Miia Hakanen (mihakane@jyu.fi) on 2022-06-21T08:47:33Z\nNo. of bitstreams: 0", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2022-06-21T08:47:33Z (GMT). No. of bitstreams: 0\n Previous issue date: 2022", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "71", "language": "", "element": "format", "qualifier": "extent", "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.title", "value": "Konjugatiivisten plasmidien is\u00e4nt\u00e4bakteerien tunnistaminen heterogeenisest\u00e4 bakteeriyhteis\u00f6st\u00e4 yksisolu-ddPCR-menetelm\u00e4n avulla", "language": "", "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-202206213526", "language": "", "element": "identifier", "qualifier": "urn", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Master\u2019s thesis", "language": "en", "element": "type", "qualifier": "ontasot", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Pro gradu -tutkielma", "language": "fi", "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": "Jyv\u00e4skyl\u00e4n yliopisto", "language": "fi", "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "University of Jyv\u00e4skyl\u00e4", "language": "en", "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": "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": "restrictedAccess", "language": null, "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": "", "element": "subject", "qualifier": "oppiainekoodi", "schema": "dc"}, {"key": "dc.subject.yso", "value": "bakteerit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "geenit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "resistenssi", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "antibioottiresistenssi", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.rights.url", "value": "https://rightsstatements.org/page/InC/1.0/", "language": null, "element": "rights", "qualifier": "url", "schema": "dc"}, {"key": "dc.rights.accessrights", "value": "The author has not given permission to make the work publicly available electronically. Therefore the material can be read only at the archival workstation at Jyv\u00e4skyl\u00e4 University Library (https://kirjasto.jyu.fi/collections/archival-workstation).", "language": "en", "element": "rights", "qualifier": "accessrights", "schema": "dc"}, {"key": "dc.rights.accessrights", "value": "Tekij\u00e4 ei ole antanut lupaa avoimeen julkaisuun, joten aineisto on luettavissa vain Jyv\u00e4skyl\u00e4n yliopiston kirjaston arkistoty\u00f6semalta. Ks. https://kirjasto.jyu.fi/kokoelmat/arkistotyoasema..", "language": "fi", "element": "rights", "qualifier": "accessrights", "schema": "dc"}]
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