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Fig. 2 | Microbial Cell Factories

Fig. 2

From: Combining manipulation of integration loci and secretory pathway on expression of an Aspergillus niger glucose oxidase gene in Trichoderma reesei

Fig. 2

The integration loci were important for expression of AnGOx in T. reesei. A: The plasmid maps of pGOx_cbh1 and pGOx_cel3c. B: Schematic diagram showing the DNA fragments that were amplified in diagnostic PCR verification of transformants with integration in the cbh1 and cel3c loci, respectively. The primer pairs used were cbh1-VF/VR and cel3c-VF/VR for integration in cbh1 and cel3c, respectively. C. PCR verification of targeted integration of the AnGOx-expressing cassette in the cbh1 and cel3c loci. The primer pair cbh1-VF/VR was used for lane 1 and 2, while the primer pair cel3c-VF/VR was used for lane 3 and 4. Lane 1: QM9414; 2: QM9414-cbh1; 3: QM9414-cel3c; 4: QM9414. M: DNA molecular mass marker. D: SDS-PAGE of the extracellular proteins of the T. reesei parent strain QM9414 and the cbh1- and cel3c-integrated AnGOx-expressing strains. Lane M: protein molecular mass marker; 1: QM9414; 2: QM9414-cbh1; 3:QM9414-cel3c. E: Time course analysis of the AnGOx activity in the supernatant of the cbh1 (QM9414-cbh1) and cel3c-integrated (QM9414-cel3c) AnGOx-expressing strains. F: Time course analysis of the AnGOx activity in the strains of QM9414-cel3c and QM9414-cel3c-cbh1 (with AnGOx integrated in both cbh1 and cel3c) strains

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Microbial Cell Factories

ISSN: 1475-2859

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