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Analysis of the transformants obtained via I- SceI-mediated gene targeting showed that about two thirds of the transformants resulted from a homologous recombination event at the predetermined locus.
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After selection on the appropriate selective plates, transformation frequencies were determined by colony counting and the calculated transformation efficiencies are shown in Tables 1 and 2. Transformants resulting from chromosomal DNA or integration vectors that were supposed to recombine at the amylose degradation amyE locus were checked for correct integration by testing for amylase activity.
Same type of correlation existed for transformants resulting from re-ligated plasmid vectors.
Transformants resulting from gene replacement at the AOX1 locus have methanol utilization slow phenotype (MutS) and can be easily identified by replica-plating on minimal methanol plates.
As shown in Figure 2A, PCR amplification of the genomic DNA obtained from primary transformants resulted in the synthesis of a product with expected 1.65 kb size.
Filament elongation was reduced in the transformants, resulting in the heterostyly type of floral architecture.
Instability of transformants usually results from a lack of integration of a given DNA cassette into the genome and has been previously described to be a major bottleneck for T. reesei transformation (Jørgensen et al. 2014).
The mRNA sequence of the transformant resulted in 100% homology with the G. graminis lox gene.
The variation observed resulted from fermentation conditions rather than differences in the genetic background of the transformants.
Transformation resulted in 44 Ferrara and 23 Ahlburg transformants.
Meiotic offspring resulting from crosses between transformants and the wild type were isolated.
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