In vivo observation of large foreign DNA molecules in host plant cells.
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.
Gisel
A
A
Institute of Plant Sciences, Swiss Federal Institute of Technology, Zürich, Switzerland.
Rothen
B
B
Iglesias
V A
VA
Potrykus
I
I
Sautter
C
C
D001483:Base Sequence; D015342:DNA Probes; D004274:DNA, Recombinant / Q000032:analysis*; D003856:Deoxyuracil Nucleotides; D005452:Fluoresceins; D018014:Gene Transfer Techniques; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D016133:Polymerase Chain Reaction; D011523:Protoplasts / Q000648:ultrastructure; D014026:Nicotiana / Q000235:genetics* / Q000648:ultrastructure
D015342:DNA Probes; D004274:DNA, Recombinant; D003856:Deoxyuracil Nucleotides; D005452:Fluoresceins; C069632:fluorescein-11-deoxyuridine triphosphate