title: Cloning the bacterial bphC gene into Nicotiana tabacum to improve the efficiency of phytoremediation of polychlorinated biphenyls.
abstract: The aim of this work was to construct transgenic plants with increased capabilities to degrade organic pollutants, such as polychlorinated biphenyls. The environmentally important gene of bacterial dioxygenase, the bphC gene, was chosen to clone into a plant of Nicotiana tabacum. The chosen bphC gene encodes 2,3-dihydroxybiphenyl-1,2-dioxygenase, which cleaves the aromatic ring of dihydroxybiphenyl, and we cloned it in fusion with the gene for β-glucuronidase (GUS), luciferase (LUC) or with a histidine tail. Several genetic constructs were designed and prepared and the possible expression of desired proteins in tobacco plants was studied by transient expression. We used genetic constructs successfully expressing dioxygenase's genes we used for preparation of transgenic tobacco plants by agrobacterial infection. The presence of transgenic DNA , mRNA and protein was determined in parental and the first filial generation of transgenic plants with the bphC gene. Properties of prepared transgenic plants will be further studied.
authors.affiliation: ICT Prague, Faculty of Food and Biochemical Technology, Department of Biochemistry and Microbiology, Prague, Czech Republic.
authors.forename: Martina
authors.initials: M
authors.lastname: Novakova
authors.forename: Martina
authors.initials: M
authors.lastname: Mackova
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Antosova
authors.forename: Jitka
authors.initials: J
authors.lastname: Viktorova
authors.forename: Miklos
authors.initials: M
authors.lastname: Szekeres
authors.forename: Katerina
authors.initials: K
authors.lastname: Demnerova
authors.forename: Tomas
authors.initials: T
authors.lastname: Macek
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001673:Biodegradation, Environmental; D042521:Burkholderiaceae / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D049308:Dioxygenases / Q000235:genetics* / Q000378:metabolism*; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D011078:Polychlorinated Biphenyls / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D011078:Polychlorinated Biphenyls; D049308:Dioxygenases; C074272:2,3-dihydroxybiphenyl oxygenase; D008156:Luciferases; D005966:Glucuronidase