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Enhancement of display efficiency in yeast display system by vector engineering and gene disruption

Abstract

Vector engineering and gene disruption in host cells were attempted for the enhancement of α-agglutinin-based display of proteins on the cell surface in yeast. To evaluate the display efficiency by flow cytometric analysis, DsRed-monomer fused with FLAG-tag was displayed and immunostained as a model protein. The use of leu2-d in the expression vector resulted in the enhanced efficiency and ratio of the accessible display of proteins. Moreover, the amount of displayed proteins in SED1-disrupted cells increased particularly during the stationary growth phase. The combination of these improvements resulted in the quantitatively enhanced accessible display of DsRed-monomer on the yeast cell surface. The improved yeast display system would be useful in a wider range of its applications in biotechnology.

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Correspondence to Mitsuyoshi Ueda.

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Kuroda, K., Matsui, K., Higuchi, S. et al. Enhancement of display efficiency in yeast display system by vector engineering and gene disruption. Appl Microbiol Biotechnol 82, 713–719 (2009). https://doi.org/10.1007/s00253-008-1808-4

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Keywords

  • Cell surface engineering
  • SED1
  • leu2-d
  • DsRed-monomer
  • Display efficiency