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[{"key": "dc.contributor.advisor", "value": "Sironen, Anu", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Yl\u00e4nne, Jari", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Sahinaho, Ulla-Maria", "language": "", "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2020-12-28T08:30:49Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2020-12-28T08:30:49Z", "language": null, "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2020", "language": "", "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/73441", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Miesten hedelm\u00e4tt\u00f6myys on kasvava ongelma l\u00e4nsimaissa. V\u00e4rekarvasairaudet voivat aiheuttaa miesten hedelm\u00e4tt\u00f6myytt\u00e4. Siitti\u00f6iden kehityksen tutkimus voi valaista syit\u00e4 siitti\u00f6iden rakenteellisten ja toiminnallisten vikojen taustalla. Sperm flagellar protein 2 (Spef2) -geeni ilmentyy kudoksissa, joissa on liikkuvia v\u00e4rekarvoja, ja se on v\u00e4ltt\u00e4m\u00e4t\u00f6n siitti\u00f6n h\u00e4nn\u00e4n kehitykselle ja liikkuvuudelle. Spef2-knockout hiirimallin avulla tutkittiin Spef2:n toimintaa v\u00e4rekarvoissa ja siitti\u00f6iss\u00e4. Liikkuvien v\u00e4rekarvojen toimintah\u00e4iri\u00f6t aiheuttavat hiirimallilla kroonisia hengitystieinfektioita, siitti\u00f6vioista johtuvaa hedelm\u00e4tt\u00f6myytt\u00e4 ja vesip\u00e4\u00e4t\u00e4. T\u00e4m\u00e4n tutkimuksen tavoite oli kehitt\u00e4\u00e4 vasta-aine, joka sitoutuu SPEF2:n adenylaattikinaasidomeeniin (KAD). Antigeeni vasta-ainetta varten tuotettiin E.colissa ja puhdistettiin affiniteettikromatografialla. Western blot-menetelm\u00e4ll\u00e4 ekspressio havaittiin kiveksess\u00e4, lis\u00e4kiveksess\u00e4, keuhkoissa, munanjohtimessa ja mahdollinen uusi isoformi aivoissa. Kaksi SPEF2-isoformia puuttui poistogeenisten hiirten kiveksist\u00e4. Kudosv\u00e4rj\u00e4yksiss\u00e4 SPEF2 lokalisoitui spermatosyytteihin, spermatideihin, spermatidien mansettiin, siitti\u00f6n h\u00e4nt\u00e4\u00e4n, Sertolin soluihin ja henkitorven epiteelisolujen v\u00e4rekarvoihin. Toinen tavoite oli kehitt\u00e4\u00e4 fluoresoiva solulinja, jolla tutkitaan SPEF2:n kuljetusta v\u00e4rekarvaan. Kolme SPEF2-GFP fuusioproteiinia ilmennettiin eri mIMCD3-solulinjoissa. 5\u00b4TE-GFP, KAD-GFP ja 3\u00b4TE-GFP vastasivat kivesspesifisen Spef2-sekvenssin eri toiminnallisia alueita. Fuusioproteiinien paikantuminen v\u00e4rekarvaan tai siihen liittyviin rakenteisiin, kuten basal bodyyn, viittaa ett\u00e4 Spef2:lla olisi useita v\u00e4rekarvalokalisaatiosignaaleja.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Male infertility is an increasing problem in Western countries. Male infertility is associated with ciliary dyskinesias, which are diseases of motile cilia. Research on sperm cell development can elucidate causes behind sperm structural and functional defects. Sperm flagellar protein 2 (Spef2) gene is expressed in ciliated tissues, and the mutations in Spef2 cause male infertility. Spef2 mutant mice have defective cilia in respiratory tract, males are infertile, and the mice develope postnatally a severe hydrocephalus causing lethality. First aim of this study was to develop a novel antibody against adenylate kinase domain (KAD) of SPEF2 protein. Antigen was produced in E. Coli and purified by affinity chromatography. In Western blot, SPEF2 isoforms were detected in wild-type testis, epididymis, lung and oviduct, and a possible novel isoform was observed in brain. Two SPEF2 isoforms were absent in knockout testis. In tissue staining, SPEF2 localized to late spermatocytes, round spermatids, manchette of elongating spermatids, sperm tail midpiece, Sertoli cells, and tracheal epithelial cilia. Second aim of the study was to create a fluorescent cell line to investigate transport of SPEF2 to the cilium and ciliary localization signals. Three SPEF2-GFP fusion proteins were expressed in separate mIMCD3 cell lines: 5\u2019TE-GFP, KAD-GFP, and 3\u2019TE-GFP, which represent different domains of testis-specific Spef2 sequence. Each fusion protein was localized to the cilium or cilium-associated structures such as basal body, which indicates that Spef2 contains multiple ciliary localization signals.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by Paivi Vuorio (paelvuor@jyu.fi) on 2020-12-28T08:30:48Z\nNo. of bitstreams: 0", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2020-12-28T08:30:49Z (GMT). No. of bitstreams: 0\n Previous issue date: 2020", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "63", "language": "", "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": "cilia", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "intraflagellar transport", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.title", "value": "Sperm flagellar protein 2 expression in mouse tissues and transfection in mIMCD3 cells", "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-202012287373", "language": "", "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": "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.collaborator", "value": "public", "language": "", "element": "contractresearch", "qualifier": "collaborator", "schema": "yvv"}, {"key": "yvv.contractresearch.funding", "value": "5000", "language": "", "element": "contractresearch", "qualifier": "funding", "schema": "yvv"}, {"key": "yvv.contractresearch.initiative", "value": "student", "language": "", "element": "contractresearch", "qualifier": "initiative", "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": 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": "sperma", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "hiiret", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "hedelm\u00e4tt\u00f6myys", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "siitti\u00f6t", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "kuljetusproteiinit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "vasta-aineet", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "sperm", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "mice", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "infertility", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "spermatozoa", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "transport proteins", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "antibodies", "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"}]
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