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After transcript analysis in these transformants, it was concluded that there was a clear correlation between transcript level and GFP accumulation.
Upon analysis of the GFP-expressing transformants, it was discovered when the fluorescent signal was present in a hyphen, it was not localized in a particular organelle or at the hyphal tip but evenly distributed.
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For transformants stabilization it was used YPD medium: 1% yeast extract, 2% peptone, 2% glucose.
Instead of using the antibiotic Cefotaxim, Timentin was used in transformant selection plates as it was found to be more efficient for removal of residual Agrobacterium tumefaciens after transformation.
(Only 1600 transformants screened here, is it meaningful?) 5) How will screening of very large numbers of transformants be achieved?
(Only 1600 transformants screened here, is it meaningful?) We demonstrated a large improvement with a small library, which we would argue is a positive.
Genomic PCR on independent transformants was performed, and 15 independent lines of T0 tobacco transformants were obtained.
The plasmid complement of transformants was verified by PCR and double restriction digestion.
Selection for double transformants was performed on –Trp Leu SD media.
One of the Ura+ transformants was named strain TR2-7.
Gene integration into prototrophic transformants was verified by genomic PCR.
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