In vivo observation of large foreign DNA molecules in host plant cells

Eur J Cell Biol. 1996 Apr;69(4):368-72.

Abstract

We have developed a system to monitor microscopically the fate of foreign DNA within plant cells in vivo. Fluorescein-11-dUTP was used to label DNA during target-specific amplification by polymerase chain reaction (PCR). Labeled DNA fragments of 1.5-3.5 kb were prepared and then transported into tobacco protoplasts by polyethylene glycol (PEG)-mediated direct gene transfer. We localized the foreign, labeled DNA within the cell by confocal laser scanning microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Probes
  • DNA, Recombinant / analysis*
  • Deoxyuracil Nucleotides
  • Fluoresceins
  • Gene Transfer Techniques
  • Microscopy, Confocal*
  • Molecular Sequence Data
  • Nicotiana / genetics*
  • Nicotiana / ultrastructure
  • Plants, Toxic*
  • Polymerase Chain Reaction
  • Protoplasts / ultrastructure

Substances

  • DNA Probes
  • DNA, Recombinant
  • Deoxyuracil Nucleotides
  • Fluoresceins
  • fluorescein-11-deoxyuridine triphosphate