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Schwarzhans, Jan Philipp; Wibberg, Daniel; Winkler, Anika; Luttermann, Tobias; Kalinowski, Jörn; Friehs, Karl: Non-canonical integration events in Pichia pastoris encountered during standard transformation analysed with genome sequencing. In: Scientific Reports. Jg.6 H. 1. 2016
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Identification of clones with abnormal colony morphology.
Figure 1. Colony morphology of wildtype P.
Table 1. Characteristics of P.
Gene disruptions.
Figure 2. The different kinds of integration events discovered in sequenced P.
Table 2. Primers used for PCR assays and qRT-PCR.
Co-integration of E. coli DNA.
Figure 3. Proposed mechanism facilitating the relocation of the AOX1 locus as found in clone JPS014.
Figure 4. Observed modification of E.
Discussion
Figure 5. Relative transcriptional levels of E.
Methods
Strains and vector.
Cultivation conditions.
Figure 6. Schematic depiction of the proposed mechanism facilitating integration of E.
Identification of Mut-phenotype and colony morphology.
Isolation of genomic DNA and PCR assays.
Genome sequencing and bioinformatic analysis of P. pastoris strains.
Genome sequencing, annotation and sequence comparison of the E. coli KRX genome.
qRT-PCR experiments.
Acknowledgements
Author Contributions