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Subsequently, potato chloroplast transformants are generated at 10 30 times lower frequencies than tobacco [ 31].

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.

A total of 117 independent transformants were generated over a period of two years.

In an attempt to isolate a further improved itaconic acid producing strains, new transformants were generated.

Multiple overexpression transformants were generated and PCR of the target genes were performed to confirm stable transformation.

Stable transformants were generated with the CfTPS2 and CfTPS3 operon in all three sites, to generate strains TPS-P, TPS-N and TPS-S (see Table 1 ).