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Similarly, using the same criterion to distinguish clustered mutations in both datasets (≥5 linked mutations separated from their neighbour by <8.5 kb), 274 of the 1064 mutations observed in AID* wild type transformants are found within clusters compared to 28 of the 2088 mutations in the AID* ung1Δ transformants (Supplementary file 1B).
While transformants were found after transformation with DNA and PCR products from the BZ586 claR strain, none of the transformants were selected after transformation with DNA from the claS strain 26695 or TE.
Our TALEN approach appears to be highly efficient: targeted mutation events were detected in 50% of all transformants obtained, whereas 21% of the transformants were found to be bi-allelic knockout lines.
The Ade+ transformants were found to contain two new YACs of 490 and 100 kb, and to lack the original 590 kb YAC, consistent with the expected splitting event.
The resulted transformants were found to be capable of growing and producing ethanol on a minimal medium at 48 °C supplemented with birchwood xylan as a sole carbon source.
The isolation of these ts mutants depended both on the earlier demonstration that homologous recombination can occur between an introduced replacement vector and the genome [3], [4] and on the unusually high level of homologous recombination which was found at this site: ∼95% of isolated transformants were found to be homologous replacements [2].
All the laccase producing transformants were found to be Mut+ (methanol utilization phenotype).
About 6.5% of diploid transformants were found to carry recessive mutations unlinked to the gene deletion.
The transformants were found to grow 20%% faster than wild-type plants (Kawaoka et al. 1994).
All PaGlo1_OEx and PaGlo2_OEx transformants were found to contain significantly elevated levels of PaGLO1 and PaGLO2, respectively.
However, since the obtained transformants all had the early leaf death phenotype and no wild type transformants were found, this possibility seems unlikely.
