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Display of heterologous proteins on the Saccharomyces cerevisiae surface display system using a single constitutive expression vector
Author(s) -
Sun Hui,
Wang Tiantian,
Zhang Jie,
Liu Qing,
Wang Longjiang,
Chen Peipei,
Wang Fangkun,
Li Hongmei,
Xiao Yihong,
Zhao Xiaomin
Publication year - 2013
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.1846
Subject(s) - heterologous , saccharomyces cerevisiae , biology , expression vector , mutant , heterologous expression , green fluorescent protein , gene , genetics , recombinant dna
In this study, we constructed a novel and simple yeast surface display system with a single expression vector. The newly established system uses a bidirectional expression vector carrying the AGA 1 gene driven by the PGK 1 promoter in one direction and the AGA 2‐expression cassette driven by the TEF 1 promoter in the reverse direction, and uses the geneticin, a G418‐resistant gene, as the selection marker for transformants. Because all the display elements are put into one expression vector, the new system is much simpler to use, and there is no need for any genetic modification of the host strains; therefore, the new system can be used in wild type as well as laboratory strains of Saccharomyces cerevisiae . The display efficiency of heterologous proteins using the new system has been confirmed by displaying enhanced green fluorescent protein and Eimeria tenella (a chicken protozoan parasite) microneme protein2 (EtMic2) on several S. cerevisiae strains. We also tested the new system with an aga 2 mutant strain of S. cerevisiae . The results indicate that the native expressed Aga2 protein has no effect on the display efficiency of heterologous proteins. © 2013 American Institute of Chemical Engineers Biotechnol. Prog ., 30:443–450, 2014