. . "Kotrba, Pavel" . "T\u0159ebo\u0148" . "22330" . . . "Bakteri\u00E1ln\u00ED transport\u00E9ry t\u011B\u017Ek\u00FDch kov\u016F a jejich potenci\u00E1l vyu\u017Eit\u00ED ve fytoremedia\u010Dn\u00EDch technologi\u00EDch"@cs . . "2011-10-17+02:00"^^ . "187767" . "P(1M06030), Z(MSM6046137305)" . "Inovativn\u00ED sana\u010Dn\u00ED technologie ve v\u00FDzkumu a praxi IV" . "Bakteri\u00E1ln\u00ED transport\u00E9ry t\u011B\u017Ek\u00FDch kov\u016F a jejich potenci\u00E1l vyu\u017Eit\u00ED ve fytoremedia\u010Dn\u00EDch technologi\u00EDch" . "[CCE28C970AB1]" . "Mackov\u00E1, Martina" . "1804-9966" . . "Vodn\u00ED zdroje Ekomonitor spol.s r.o." . . . . . . "5"^^ . . . "Chrudim" . "\u0160uman, J\u00E1chym" . "Nov\u00E1kov\u00E1, Martina" . "Pr\u00E1ce byla zam\u011B\u0159ena na studium bakteri\u00E1ln\u00EDch gen\u016F metA, metTs a pbrT k\u00F3duj\u00EDc\u00EDch transport\u00E9ry iont\u016F t\u011B\u017Ek\u00FDch kov\u016F. Geny metA a metTs lokalizuj\u00ED na megaplasmidu pA81 v Gramnegativn\u00ED p\u016Fdn\u00ED bakterii Achromobacter xylosoxidans A8 a k\u00F3duj\u00ED P1-ATPasu exportuj\u00EDc\u00ED ionty Cd2+ a Zn2+ z cytoplasmy do periplasmy a hypotetick\u00FD proteinov\u00FD transport\u00E9r iont\u016F t\u011B\u017Ek\u00FDch kov\u016F, v uveden\u00E9m po\u0159ad\u00ED. Gen pbrT je sou\u010D\u00E1st\u00ED determinanty pbr p\u016Fvodem z multiresistentn\u00ED bakterie Cupriavidus metallidurans CH34 a k\u00F3duje transmembr\u00E1nov\u00FD protein transportuj\u00EDc\u00ED ionty Pb2+ z periplasmy bakteri\u00E1ln\u00ED bu\u0148ky do cytoplasmy. Za \u00FA\u010Delem zji\u0161t\u011Bn\u00ED jejich fenotypov\u00E9ho efektu v eukaryotn\u00EDm modelu, byla konstruov\u00E1na s\u00E9rie plasmid\u016F umo\u017E\u0148uj\u00EDc\u00ED konstitutivn\u00ED expresi gen\u016F metA, metTs a pbrT v kvasink\u00E1ch Saccharomyces cerevisiae. Bylo prok\u00E1z\u00E1no, \u017Ee heterologn\u00ED exprese genu metA v bu\u0148k\u00E1ch kmene S. cerevisiae DTY168 (kmen hypersensitivn\u00ED k Cd2+ a Pb2+) zvy\u0161uje jejich sensitivu k Cd2+. Z p\u0159edb\u011B\u017En\u00FDch v\u00FDsledk\u016F d\u00E1le vypl\u00FDv\u00E1, \u017Ee exprese gen\u016F metA a metTs zvy\u0161uje akumulaci iont\u016F Cd2+ a Zn2+ v bu\u0148k\u00E1ch tohoto kmene. Studov\u00E1na byla t\u00E9\u017E subcelul\u00E1rn\u00ED lokalizace proteinov\u00FDch produkt\u016F studovan\u00FDch gen\u016F. Byla p\u0159ipravena s\u00E9rie plasmid\u016F umo\u017E\u0148uj\u00EDc\u00EDch expresi gen\u016F metA, metTs a pbrT ve f\u00FAzi s genem k\u00F3duj\u00EDc\u00EDm zelen\u00FD fluorescen\u010Dn\u00ED protein eGFP. Pomoc\u00ED fluorescen\u010Dn\u00ED mikroskopie bylo prok\u00E1z\u00E1no, \u017Ee proteinov\u00E9 produkty f\u00FAzn\u00EDch gen\u016F v bu\u0148k\u00E1ch S. cerevisiae lokalizuj\u00ED v cytoplasmatick\u00E9 membr\u00E1n\u011B a v tonoplastu." . . "5"^^ . "The work was focused on the study of bacterial metA, metTs and pbrT genes encoding for heavy metal ions transporters. MetA and metTs genes are originally localized on pA81 megaplasmid in Gramnegative soil bacterium Achromobacter xylosoxidans A8 and they encode for P1-ATPase transporting Cd2+/Zn2+ ions from cytoplasm to periplasm and putative heavy metal transporter, respectively. PbrT gene constitutes pbr determinant carried by multi-metalloresistant bacterium Cupriavidus metallidurans CH34 and it encodes for transmembrane protein transporting Pb2+ ions from periplasm of bacterial cell to cytoplasm of bacterial cell. In order to reveal their phenotypical effect in eukaryotic model a set of plasmids enabling constitutive expression of met and pbrT genes in yeasts was constructed. It was demonstrated, that constitutive expression of metA gene in Saccharomyces cerevisiae strain DTY168 (Cd2+/Pb2+-hypersensitive strain) increases Cd2+ sensitivity of transformed cells. According to preliminary results, metA and metTs expression also increases intracellular Cd/Zn accumulation. Subcellular localization of met and pbrT protein products was also studied. A set of expression plasmids carrying met and pbrT coding sequences in 3'-fusion with green fluorescent protein (eGFP) gene was prepared. Using fluorescent microscopy it was demonstrated that proteins expressed from all three fusion genes localize in cytoplasmic membrane and tonoplast of S. cerevisiae cells."@en . . "RIV/60461373:22330/11:43891894!RIV12-MSM-22330___" . . . "RIV/60461373:22330/11:43891894" . "Bakteri\u00E1ln\u00ED transport\u00E9ry t\u011B\u017Ek\u00FDch kov\u016F a jejich potenci\u00E1l vyu\u017Eit\u00ED ve fytoremedia\u010Dn\u00EDch technologi\u00EDch" . "Macek, Tom\u00E1\u0161" . . "Bacterial heavy metal transporters and their potential for use in fytoremediations"@en . . . "bioremediation; genetically modified organisms; Saccharomyces cerevisiae; heavy metal transporters; bacterial metalloresistance; heavy metals"@en . "5"^^ . . . "Pr\u00E1ce byla zam\u011B\u0159ena na studium bakteri\u00E1ln\u00EDch gen\u016F metA, metTs a pbrT k\u00F3duj\u00EDc\u00EDch transport\u00E9ry iont\u016F t\u011B\u017Ek\u00FDch kov\u016F. Geny metA a metTs lokalizuj\u00ED na megaplasmidu pA81 v Gramnegativn\u00ED p\u016Fdn\u00ED bakterii Achromobacter xylosoxidans A8 a k\u00F3duj\u00ED P1-ATPasu exportuj\u00EDc\u00ED ionty Cd2+ a Zn2+ z cytoplasmy do periplasmy a hypotetick\u00FD proteinov\u00FD transport\u00E9r iont\u016F t\u011B\u017Ek\u00FDch kov\u016F, v uveden\u00E9m po\u0159ad\u00ED. Gen pbrT je sou\u010D\u00E1st\u00ED determinanty pbr p\u016Fvodem z multiresistentn\u00ED bakterie Cupriavidus metallidurans CH34 a k\u00F3duje transmembr\u00E1nov\u00FD protein transportuj\u00EDc\u00ED ionty Pb2+ z periplasmy bakteri\u00E1ln\u00ED bu\u0148ky do cytoplasmy. Za \u00FA\u010Delem zji\u0161t\u011Bn\u00ED jejich fenotypov\u00E9ho efektu v eukaryotn\u00EDm modelu, byla konstruov\u00E1na s\u00E9rie plasmid\u016F umo\u017E\u0148uj\u00EDc\u00ED konstitutivn\u00ED expresi gen\u016F metA, metTs a pbrT v kvasink\u00E1ch Saccharomyces cerevisiae. Bylo prok\u00E1z\u00E1no, \u017Ee heterologn\u00ED exprese genu metA v bu\u0148k\u00E1ch kmene S. cerevisiae DTY168 (kmen hypersensitivn\u00ED k Cd2+ a Pb2+) zvy\u0161uje jejich sensitivu k Cd2+. Z p\u0159edb\u011B\u017En\u00FDch v\u00FDsledk\u016F d\u00E1le vypl\u00FDv\u00E1, \u017Ee exprese gen\u016F metA a metTs zvy\u0161uje akumulaci iont\u016F Cd2+ a Zn2+ v bu\u0148k\u00E1ch tohoto kmene. Studov\u00E1na byla t\u00E9\u017E subcelul\u00E1rn\u00ED lokalizace proteinov\u00FDch produkt\u016F studovan\u00FDch gen\u016F. Byla p\u0159ipravena s\u00E9rie plasmid\u016F umo\u017E\u0148uj\u00EDc\u00EDch expresi gen\u016F metA, metTs a pbrT ve f\u00FAzi s genem k\u00F3duj\u00EDc\u00EDm zelen\u00FD fluorescen\u010Dn\u00ED protein eGFP. Pomoc\u00ED fluorescen\u010Dn\u00ED mikroskopie bylo prok\u00E1z\u00E1no, \u017Ee proteinov\u00E9 produkty f\u00FAzn\u00EDch gen\u016F v bu\u0148k\u00E1ch S. cerevisiae lokalizuj\u00ED v cytoplasmatick\u00E9 membr\u00E1n\u011B a v tonoplastu."@cs . "978-80-86832-61-6" . . "Bacterial heavy metal transporters and their potential for use in fytoremediations"@en . . . "Bakteri\u00E1ln\u00ED transport\u00E9ry t\u011B\u017Ek\u00FDch kov\u016F a jejich potenci\u00E1l vyu\u017Eit\u00ED ve fytoremedia\u010Dn\u00EDch technologi\u00EDch"@cs . .