After recieving a GFP-construct in a Ti-plasmid, we are transiently transfecting GFP plants.
The recombinant plasmid is then put into agrobacterium (by heat shock transformation) and the bacteria are cultivated in liquid medium (plus selection antibiotic). After two or three days, the bacteria have propagated enough, are centrifuged and resuspended in fresh LB medium, and they ar ethen put onto a plant. After 5-7 days, the plant shold look like this:
Thanks to T. Moravec, IEB, Prague for providing the fotos.
Everywhere, where agrobacterium touched the plants, it has inserted the gene for the green fluorescent protein into the plants genome, and the plant emits light from these spots under UV light. (The bacteria afterwards are killed of with ethanol or antibiotic or bleach)
Thus the plants are not really glowing yet, because you need UV light. But a nice show object where the journey is going. And in this particular case, it’s a genetic mosaic plant, as by far not every plant cell contains the new gene.
The pGlowroplast backbone hopefully will arrive soon from synthesis, then itÄ’s about engineering real glowing plants!