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In order to characterize the extent and nature of multiple vector transformants in this important host, plasmid-born gene libraries constructed by yeast homologous recombination were analyzed by DNA sequencing.

Fig. 4 Assessment of genetic stability of transformants in the absence of selective pressure.

PCR screening using adaptor primer and VRV (CT 15 as primers identified 104 microsatellite clones from 422 transformants in the libraries.

The constructed vector was stably maintained in the transformants, in the presence or absence of butanol stress.

Figure 2 Correlation between the level of transcription of the aphII gene and the survival rates of different transformants in the presence of 100 μg.ml -1 neomycin.

The transformants in the present study produce the highest total IBE concentration reported for clostridial batch cultures without online IBE removal (24.4 g /L).

The sequence-verified constructs contained a kanamycin resistance cassette, allowing screening for positive transformants in the T1 generation.

Transformants in the irx7 and irx8 backgrounds were grown entirely with a 16 h photoperiod.

Transformants in the irx7 background harboring the pVND6 IRX7 construct had a significantly lower lignin content compared to the wild type, whereas the other transformants in irx7 and irx8 mutant backgrounds did not show significant changes in lignin content.

Moreover, growth of all 38 transformants in the presence of antibiotic already demonstrated the functional expression of the aphVIII gene.

We were only able to obtain pure, homokaryotic transformants in the case of the ΔhacA transformants (compact growing colonies), whereas propagation of the ΔireA transformants was only possible when substrate mycelium was transferred (data not shown).