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. 2018 Sep 27;9:2300. doi: 10.3389/fmicb.2018.02300

FIGURE 1.

FIGURE 1

Schematic construct of open reading frames (OFRs), expression cassette, and PCR amplicons. (A) ORF construct for proform PG-1, cathelin, and mature PG-1 expression. MeOH-inducible AOX1 promoter and α-mating factor secretion signal sequence was used in all expression constructs. Asterisk (∗) indicate enterokinase (EK) cleavage site. (B) Schematic representation of the integration of the expression cassette into Pichia pastoris genome. (C) PCR products amplified from gDNA to confirm integration of constructs for the expression of rProPG-1 (lane 1, 2 from two representative positive clones, respectively, at 760 bp) and rPG-1 (lane 4, 5 from two representative positive clones, respectively, at 458 bp). Expression plasmids for rProPG-1 and rPG-1 were used as an assay control (lane 3 and 6, respectively). (D) PCR products amplified from gDNA to confirm integration of constructs for the expression of rCath (lane 1–5). Expression plasmids for rCath were used as an assay control (lane 6). Wild-type AOXI gene yields a 2.2 kb product (Lane 7). Scales are not proportional to the sizes of the elements in (A) and (B).