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Bioresources and Bioprocessing
Fig. 1 | Bioresources and Bioprocessing

Fig. 1

From: High-efficiency secretory expression and characterization of the recombinant type III human-like collagen in Pichia pastoris

Fig. 1

A The construction process of hlCOLIII high-yielding strain. The target gene after hydrophobic amino acid substitution was inserted into the pPIC9K expression vector and introduced into GS115. The transformants grown on the MD screening plate were subjected to screening for multicopy strains on plates containing high concentrations of G418. B SDS-PAGE analysis of 9 recombinant strains selected from the plate containing high concentration of G418 (5 mg mL−1). Lanes 1–9 were the expression levels of recombinant strains 1#–9# (loading volume 20 μL, 3 times concentrated by trichloroacetic acid), respectively, as well as lanes M1 and M2 were standard molecular weights. C Western blotting analysis of the expression products of 2# recombinant strain. Lane 1 was the blank control group (GS115/pPIC9K), lane 2 was the experimental group (2# GS115/pPIC9K-col), and lane M was standard molecular weights. D The expression levels of hlCOLIII were driven by 7 different promoters in cooperation with PAOX1. Taking the recombinant strain GS115/pPIC9K-col (2#) as the starting strain (control)

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