20.500.12556/RUL-96394 Priprava in analiza izražanja fuzij genov za virulentne dejavnike bakterije Escherichia coli in genov za fluorescentne proteine Construction and expression analysis of fusions of Escherichia coli virulence factor genes and genes for fluorescent proteins Sevi bakterije Escherichia coli so zelo raznoliki. Okužbe povzročajo sevi, ki so pridobili zapise za sintezo virulentnih dejavnikov. Pri komenzalih in patogenih sevih E. coli so za prvi stik z gostiteljem pomembni adhezini, za preživetje različni sistemi za prevzem železa in izogibanje imunskemu sitemu gostitelja. Patogeni bakterijski sevi imajo tudi zapisi za toksine, ki delujejo na gostiteljeve celice. V populaciji genetsko identičnih bakterijskih celic je poznan fenomen fenotipske variabilnosti. Heterogeno izražanje genov bakterijam nudi prednost v spremenljivem okolju, saj je tako del populacije pripravljen na hitre in neugodne spremembe. Namen naše raziskave je bil pripraviti genske fuzije promotorjev nekaterih genov za virulentne dejavnike in genov za fluorescentne proteine ter analizirati izražanje s fluorescentnim mikroskopom. Preučevali smo promotorje genov, clbA za fosfopanteteinil transferazo, cnf1 za toksin Cnf1 in cdtABC za citotoksin CDT. Genske fuzije smo v več korakih pripravili v plazmidnem vektorju pBAC in ga vnesli v celice bakterijskega seva MG1655. Z genskimi fuzijami smo ugotovili, da vse celice E. coli izražajo gen clbA. Ugotovili smo, da je izražanje genov za sintezo toksina Cnf1 in citotoksina CDT bakterije E. coli heterogeno, saj se izražata le v majhnem delu populacije. Escherichia coli is a very diverse bacterial species. Bacterial strains with genes for virulence factors provoke infections. For commensal and pathogenic strains, adhesins are essential for first contact with the host, they also have systems for iron uptake and to avoid the host’s immune system. Pathogenic strains, harbor genes for toxins that damage host cells. In populations of genetically identical bacterial cells, phenotypic heterogeneity has been described. Heterogeneous gene expression provides bacteria an advantage in variable environments and under stress conditions. The purpose of our study was to construct gene fusions of promoters of some virulence factor genes and genes for fluorescent proteins and subsequently analyze their expression with fluorescence microscopy. We studied promoters of genes, clbA for phosphopanteteinyl transferase, cnf1 for toxin Cnf1, and cdtABC for cytoxin CDT. Gene fusions were prepared stepwise in the plasmid vector pBAC and introduced into the bacterial strain MG1655. We found that all cells expressed the clbA gene. Expression of genes for the Cnf1 toxin and cytotoxin CDT was heterogeneous, as they were expressed in only a small subpopulation. patogene bakterije Escherichia coli virulentni dejavniki genska fuzija izražanje genov fluorescentni proteini fenotipska heterogenost fluorescentna mikroskopija pathogenic bacteria Escherichia coli virulence factors gene fusion gene expression fluorescent proteins phenotypic heterogeneity fluorescence microscopy true false false [A. Petrič] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2017-09-30 07:15:26 2017-09-30 07:15:28 2022-08-12 11:19:22 0000-00-00 00:00:00 2017 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 579.253.2:579.842:577.21 146368 4821368 1440.pdf 1440.pdf 1 8454646FDA354B045598664358B7F073 c067f1a9538c75d5ab783da4fd3fea645b83f1682bd9b8753df2082499e30567 e6405e82-a1b3-11eb-a523-00155dcfd717 20.500.12556/rul/7fcd74f3-e5cd-4e19-a456-42de3746d8cc https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=105054 Biotehniška fakulteta 0 0 0