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Transformants were generated by co-transformation with the selectable pPmr3 plasmid and one of the three gluc-derived, non-selectable reporter gene plasmids at a time.
Transformants were generated by the biolistic transformation method.
We estimated ∼30,000 transformants were generated in this library.
All the Arabidopsis T1 transformants were generated by Agrobacterium-mediated infiltration, as previously described, and selected in plate media containing 0.01% (w/v) kanamycin and 0.01% gentamycin (w/v).
Six independent germ-line transformants were generated for the hUbb+1 construct.
Stable transformants were generated via homologous recombination between the linearized plasmid DNA and genomic yeast DNA.
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Subsequently, potato chloroplast transformants are generated at 10 30 times lower frequencies than tobacco [ 31].
Transformants that were generated through co-transformation with plasmid pPsaD-GLuc were analyzed for luciferase activity.
Transformants that were generated through co-transformation with the ars plasmid ptubar4 were analyzed for arylsulfatase activity.
Transformants that were generated through co-transformation with plasmid pHsp70A-GLuc were also analyzed qualitatively for luciferase activity, and several of these transformants showed chemiluminescence (data not shown).
However, transformants that were generated using the plasmids pPsaD-GLuc and pHsp70A-GLuc showed functional expression of luciferase through light emission, which is easily detected in bioluminescence assays.
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