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PCR Cloning and Subcloning> | pastoris and synthesized as a Genparts DNA fragment by Genscript without an internal BamHI restriction site.... In-vitro multicopy hydrophobin cloning: e l c i t r A d e t p e c c A Hydrophobin HFBI gene was codon optimized for Pichia pastoris expression with N-terminal hexahistidine (His6)– hemagglutinin (HA) tags for purification and detection and synthesized by Genscript with a conservative phenylalanine to tryptophan mutation for protein quantitation by UV-Vis, as previously reported (Sallada et al. | Get A Quote |
Hydrophobins are small highly surface-active fungal proteins with potential as biosurfactants in a wide array of applications. However, practical implementation of hydrophobins at large scale has been hindered by low recombinant yields. In this study, the effects of increasing hydrophobin gene copy number and overexpressing endoplasmic reticulum resident chaperone proteins Kar2p, Pdi1p, and Ero1p were explored as a means to enhance recombinant yields of the class II hydrophobin HFBI in the eukaryotic expression host Pichia pastoris. One-, 2-, and 3-copy-HFBI strains were attained using an in vitro multimer ligation approach, with strains displaying copy number stability following subsequent transf... More