About: Targeted gene replacement of a ferredoxin gene in Trichomonas vaginalis does not lead to metronidazole resistance

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About: Targeted gene replacement of a ferredoxin gene in Trichomonas vaginalis does not lead to metronidazole resistance Goto Sponge Distinct Permalink

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rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
Description	Ferredoxin, Fd, is often deficient in metronidazole-resistant strains of Trichomonas vaginalis and is thought to be necessary for drug activation. To directly test whether Fd is essential for metronidazole susceptibility, gene replacement technology has been developed for T. vaginalis. The selectable marker gene neomycin phosphotransferase (NEO) flanked by approximately 2.6 and approximately 2.0 kBp of the Fd 5' and 3' flanking regions (pKO-FD-NEO) was introduced into cells on linear DNA and selected for NEO gene expression. Stable transformants were shown to contain the NEO gene in the Fd locus and to have completely lost the Fd gene. Northern and immunoblot analyses confirm the loss of Fd mRNA and protein in pKO-FD-NEO cells. Analyses of the activity of hydrogenosomal proteins in Fd KO cells show a fourfold increase in hydrogenase activity and a 95% decrease in pyruvate/ferredoxin oxidoreductase (PFO) activity. In contrast, PFO and hydrogenase mRNA levels are unchanged. Ferredoxin, Fd, is often deficient in metronidazole-resistant strains of Trichomonas vaginalis and is thought to be necessary for drug activation. To directly test whether Fd is essential for metronidazole susceptibility, gene replacement technology has been developed for T. vaginalis. The selectable marker gene neomycin phosphotransferase (NEO) flanked by approximately 2.6 and approximately 2.0 kBp of the Fd 5' and 3' flanking regions (pKO-FD-NEO) was introduced into cells on linear DNA and selected for NEO gene expression. Stable transformants were shown to contain the NEO gene in the Fd locus and to have completely lost the Fd gene. Northern and immunoblot analyses confirm the loss of Fd mRNA and protein in pKO-FD-NEO cells. Analyses of the activity of hydrogenosomal proteins in Fd KO cells show a fourfold increase in hydrogenase activity and a 95% decrease in pyruvate/ferredoxin oxidoreductase (PFO) activity. In contrast, PFO and hydrogenase mRNA levels are unchanged. (en)
Title	Targeted gene replacement of a ferredoxin gene in Trichomonas vaginalis does not lead to metronidazole resistance Targeted gene replacement of a ferredoxin gene in Trichomonas vaginalis does not lead to metronidazole resistance (en)
skos:prefLabel	Targeted gene replacement of a ferredoxin gene in Trichomonas vaginalis does not lead to metronidazole resistance Targeted gene replacement of a ferredoxin gene in Trichomonas vaginalis does not lead to metronidazole resistance (en)
skos:notation	RIV/00216224:14310/04:00036237!RIV10-MSM-14310___
http://linked.open...avai/riv/aktivita	P S V
http://linked.open...avai/riv/aktivity	P(GA204/00/1561), S, V
http://linked.open...iv/cisloPeriodika	1
http://linked.open...vai/riv/dodaniDat	2010
http://linked.open...aciTvurceVysledku	Vaňáčová, Štěpánka
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Masarykova univerzita / Přírodovědecká fakulta
http://linked.open...dnocenehoVysledku	630236
http://linked.open...ai/riv/idVysledku	RIV/00216224:14310/04:00036237
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	Trichomonads; ferredoxin; metronidazole; drug resistance; gene knock-out (en)
http://linked.open.../riv/klicoveSlovo	drug resistance metronidazole Trichomonads ferredoxin gene knock-out
http://linked.open...odStatuVydavatele	US - Spojené státy americké
http://linked.open...ontrolniKodProRIV	[CBF081A86DC6]
http://linked.open...i/riv/nazevZdroje	Molecular Microbiology
http://linked.open...in/vavai/riv/obor	EE
http://linked.open...ichTvurcuVysledku	1 (xsd:int)
http://linked.open...cetTvurcuVysledku	7 (xsd:int)
http://linked.open...vavai/riv/projekt	Regulatory role of iron in energetic metabolism of trichomonads
http://linked.open...UplatneniVysledku	2004
http://linked.open...v/svazekPeriodika	51
http://linked.open...iv/tvurceVysledku	Tachezy, Jan Vaňáčová, Štěpánka Sutak, R. Johnson, Patricia J. Dellgadillo-Correa, M. G. Hsieh, C. L. Land, K. M.
http://linked.open...ain/vavai/riv/wos	000186974100011
issn	0950-382X
number of pages	8 (xsd:int)
http://localhost/t...ganizacniJednotka	14310

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