The average rosette area for the four most robust SUC2p::cSUC2 transformants was not significantly different from heterozygous plants (Fig. 2, Table 1) [ 25].
Since pINA1297 vector has a random integration mechanism into Y. lipolytica genome [ 14], transformants are usually different in terms of the integration loci and copy number, which is expected to affect protein expression level [ 16].
To test whether the morphogenetic defects of gas1Δ cells were also complemented by gas4+, the morphology of the transformants was analyzed and the different defects were quantified.
Thus, even the extreme variability in luciferase activity between different transformants was permanent.
After proteinase K treatment, green fluorescence on the cell surface of A. oryzae/GFP-CBM and A. oryzae/CBM-GFP transformants was diminished (Fig. 3c, f), whereas the A. oryzae/GFP transformant was scarcely different before and after treatment (Fig. 3h, i).
A third plasmid, pG_sDF, was then transferred into P. p/MJ_J 9, and the transformants were screened against different concentrations of hygromycin to find lovastatin-producing strains.
Although majority of the transformants were not notably different from nontransformants, we did find two transgenic tomato plants that have unexpected flower phenotypes with leaf-like sepals (named Leafy Sepals or LS1 and LS3; Figure 1).
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.
