Figure 7
Analysis of the expression of α-MeH chimeric protein containing S. cerevisiae α-factor signal sequence. Whole cell lysates of methanol induced P. pastoris cells (Mut+ phenotype) expressing α-MeH chimeric protein containing S. cerevisiae α-factor signal sequence instead of native TM anchor domain were resolved by SDS-PAGE, blotted onto nitrocellulose membrane and analysed by Western blotting using anti-His antibody. M- protein marker, C- control (pPIC9K Mut+), VK- one-copy pPIC9K-MeH transformant, 1,2,3- multicopy pPIC9K-MeH transformants. Solid arrow indicates unglycosylated MeH polypeptides, dotted arrow - glycosylated MeH forms.