Cloning the bacterial bphC gene into Nicotiana tabacum to improve the efficiency of phytoremediation of polychlorinated biphenyls

Bioeng Bugs. 2010 Nov-Dec;1(6):419-23. doi: 10.4161/bbug.1.6.12723.

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.

Keywords: 2,3-dihydroxybiphenyl-1,2-dioxygenase; Nicotiana tabacum; bphC; phytoremediation; transgenic plant.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Biodegradation, Environmental
  • Burkholderiaceae / enzymology*
  • Burkholderiaceae / genetics
  • Cloning, Molecular
  • Dioxygenases / genetics*
  • Dioxygenases / metabolism*
  • Glucuronidase / genetics
  • Glucuronidase / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Luciferases / genetics
  • Luciferases / metabolism
  • Nicotiana / enzymology
  • Nicotiana / genetics*
  • Nicotiana / metabolism
  • Plants, Genetically Modified / enzymology
  • Plants, Genetically Modified / genetics*
  • Plants, Genetically Modified / metabolism
  • Polychlorinated Biphenyls / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Bacterial Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Polychlorinated Biphenyls
  • Dioxygenases
  • 2,3-dihydroxybiphenyl oxygenase
  • Luciferases
  • Glucuronidase