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Figure 6 | Microbial Cell Factories

Figure 6

From: Application of a wide-range yeast vector (CoMedâ„¢) system to recombinant protein production in dimorphic Arxula adeninivorans, methylotrophic Hansenula polymorpha and other yeasts

Figure 6

Glycosylation analysis of secreted IFNγ. A strain of the pMrL-IFNg/pMrL-CNE collection was cultured on a 500 ml scale. The proteins of the supernatant were precipitated with (NH4)2SO4. An aliquot of the precipitate was digested with PNGase F as described in Materials and methods. Untreated and PNGase F-treated protein samples were compared to isolates from a previous strain collection. The various samples were separated by SDS-PAGE; proteins were visualized by immunoblotting as described before. (1,5,9,13) IFNγ standard (E.coli), (2,6,10,14) H. polymorpha, (3,4) H. polymorpha/CoMed8-MATα-IFNγ, (7,8) H. polymorpha/CoMed8-MATα-IFNγ – deglycosylated, (11,12) H. polymorpha/CoMed8-MATα-IFNγ – CoMed14-CNE, (15,16) H. polymorpha/CoMed8-MATα-IFNγ – CoMed14-CNE – deglycosylated.

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Microbial Cell Factories

ISSN: 1475-2859

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