Fig. 1

Expression construct pPICZαA/BLIP for production of secretory BLIP in P. pastoris. a Plasmid map of pPICZαA/BLIP (AOX1 promoter alcohol oxidase 1 promoter that permits the methanol-inducible expression of BLIP in Pichia, BLIP terminator blip transcription terminator that allows 3′mRNA processing of blip gene, TEF1 promoter and EM7 promoter transcription elongation factor 1 gene from Saccharomyces cerevisiae and a synthetic prokaryotic promoter that drive the expression of the Zeocin resistance gene, Zeocin resistance marker Sh ble gene1 whose product confers resistance to Zeocin in Pichia cells for selection, CYC terminator 3′ end of the Saccharomyces cerevisiae cytochrome c1 gene that allows efficient 3′ mRNA processing of the Zeocin resistance gene, ori origin of replication); b Sequence that encodes the mature protein of BLIP was placed downstream the AOX1 promoter and α-factor mating signal sequence (sequence encoding mature BLIP and the blip transcription terminator were shaded; EcoRI and XbaI restriction sites were underlined; Kex2 and Ste13 signal cleavage sites were indicated by arrow head and arrow respectively)