Abstract：
:We describe here the construction of Gateway-compatible vectors, pBGP1-DEST and pPICZα-DEST, for rapid and convenient preparation of expression plasmids for production of secretory proteins in Pichia pastoris. Both vectors direct the synthesis of fusion proteins consisting of the N-terminal signal and pro-sequences of Saccharomyces cerevisiae α-factor, the recognition sites for Kex2 and Ste13 processing proteases, the mature region of a foreign protein flanked by attB1- and attB2-derived sequences at N- and C-termini, respectively, and myc plus hexahistidine tags added at the extreme C-terminus. To test the usefulness of these vectors, production of endo-glucanases and xylanases from termite symbionts, as well as a fungal glucuronoyl esterase, was performed. Enzyme activities were detected in the culture supernatants, indicating that the chimeric proteins were synthesized and secreted as designed.
journal_name
Plasmidjournal_title
Plasmidauthors
Sasagawa T,Matsui M,Kobayashi Y,Otagiri M,Moriya S,Sakamoto Y,Ito Y,Lee CC,Kitamoto K,Arioka Mdoi
10.1016/j.plasmid.2010.08.004subject
Has Abstractpub_date
2011-01-01 00:00:00pages
65-9issue
1eissn
0147-619Xissn
1095-9890pii
S0147-619X(10)00071-5journal_volume
65pub_type
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