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Southern analysis of the transformants confirmed that pBT6 exists as an integration vector, and laeA integrated in the genome at random.
In any case, we were able to transform N. europaea with pSK2 many times, and frequent plasmid preparations and PCR amplification of vgb from these transformants confirmed that pSK2, once transformed into N. europaea, was stably maintained.
The measurement of β-galactosidase activity in these transformants confirmed that MP interacts with CP, VPg and P10 but not with P8, RdRp or Pro.
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Furthermore, induction of genes we found attenuated in sak1 was restored in these transformants, confirming that Cre17.g741300 is the SAK1 gene required for acclimation and the gene expression response to O2.
Sequence analysis of the deletion transformant confirmed that the chromosomal INU1 locus harbored a 2565-bp deletion between the gRNA-2 and gRNA-1 targeting sites, yielding Δinu1 (Km02-063) strain (Fig. 1a).
Polymerase chain reaction detection of the hph-gene from DNA of five independent putative transformants confirmed integration of hph into the genome of A95 transformants.
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Ivna confirms that assessment.
DNA gel blot analysis confirmed that the transformants contained ectopic insertions of the VdGARP1-complementing construct and the original T-DNA insertion of the vdgarp1 background (Figure 4C).
Transgene integration was verified by mPCR in all primary transformants (T0), while immunoblot analysis confirmed that the protein HrpZPsph was produced and the signal peptide was properly processed.
We obtained NsbHLH2 overexpressing transformants of N. salina CCMP1776 by particle bombardment and confirmed that these were stable transformants.
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