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Iskanje genov za ABC-transportne proteine glive Rhizopus nigricans = Identification of the genes coding for ABC-transporte proteins of the funges Rhizopus nigricans : diplomska nalogaŠtrbenc, AndrejaZigomicetna filamentozna gliva Rhizopus nigricans je poznana kot organizem, ki je sposobna z razlièno uèinkovitostjo biotransformirati nekatere steroide, kot so progesteron, 21-OH progesteron in ... testosteron. Za ta organizem steroidinajverjetneje predstavljajo ksenobiotike, ki se z inducibilnim hidroksilaznim sistemom pretvorijo v manj toksiène bolj hidrofilne produkte, ki se lazje izloèijo iz organizma. Po analogiji z drugimi evkariontskimi organizmi bi lahko bili v glivi R. nigricans v proces izloèanja hidroksiliranih steroidov vkljuèeni predstavniki poddruzine transporterjev ABC, ki iz celic izloèajo organizmu tuje snovi. Za iskanje genov z zapisi za transportne proteine ABC, v glivi R. nigricans smo uporabili knjiznico cDNA, ki je bila predhodno pripravljena iz glive izpostavljene 20 min progesteronu. V taksnih pogojih se namreè poveèa kolièina mRNA za vrsto proteinov, ki pod normalnimi rastnimi pogoji niso prisotni v glivnih micelijih. DNA bakteriofagov iz plakov smo hibridizirali z radioktivno oznaèenim fragmentom DNA z zapisom za del transporterja ABC glive R. nigricans in osamili 13 pozitivnih klonov. Po pretvorbi bakteriofagne DNA v fagmidno smo na osnovi restrikcijskih map fragmentov cDNA sklepali, da smo izolirali tri razliène klone z zapisi ta transporterje ABC. Z analizami nukleotidnih zaporedij smo dokazali, da sta zaporedji cDNA klonov ABC1 (1188 bp) in ABC2 (1904 bp) identièni, razlika je le v velikostih fragmentov. Poleg tega smo pokazali, da se klon ABC4 z nukleotidnim zaporedjem dolgim 1601 bp razlikuje od prvih dveh in da je identiènost zaporedij cDNA klonov ABC2 in ABC4 58.6%. Na osnovi analiz cDNA aminokislinskih zaporedij dolgih 596 aminokislin (klon ABC2) in 493 aminokislin (klon ABC4) smo lahko tudi zakljuèili, da smo izolirali del zapisov za transporterja ABC glive R. nigricans in da jim manjka daljse oz. krajse 5ć podroèje. S primerjavami aminokislinskih zaporedij, izvedenih iz nukleotidnih zaporedij klonov ABC2 in ABC4, z zaporedji vsebovanim v podatkovni bazi GenBank, pa smo ugotovili, da je identiènost delov transportnih proteinov ABC glive R. nigricans najveèja s predstavnikom transporterjev ABC, iz poddruzine transporterjev MD glive Mucor racemosus ter znasa 62.7% oz. 41.9% (Slika 18, Tabela 2).Type of material - undergraduate thesis ; adult, seriousPublication and manufacture - Ljubljana : [Štrbenc A.], 2004Language - slovenianCOBISS.SI-ID - 1577329
Author
Štrbenc, Andreja
Other authors
Črešnar, Bronislava, 1950-2024
Topics
Rhizopus nigricans |
cDNA knjižnica |
transportni proteini |
hibridizacija
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Faculty of Pharmacy, Ljubljana | Ljubljana | FFALJ |
reading room 1 cop.
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Štrbenc, Andreja | ![]() |
Črešnar, Bronislava, 1950-2024 | 08777 |
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