Figure 5
Expression of His-tagged MeH in P. pastoris transformants induced with methanol. (A) SDS-PAGE of whole cell lysates from P. pastoris transformants carrying empty vector or multicopy MeH expression cassetes. (B) Western blotting of the same samples using anti-tetraHis antibody. M- prestained protein marker, C- Control (pPIC3.5K MutS), 1,2,3- multicopy pPIC3.5K-MeH transformants for intracellular expression of MeH, resistant to various concentrations of antibiotic G418. Solid arrows indicate unglycosylated MeH polypeptide precursor, dotted arrows point to glycosylated MeH form and dashed arrows show cellular proteins, upregulated in response to MeH expression.