A versatile vector was developed for heterologous proteins display on the cell surface of Pichia pastoris using the C-terminal half of α-agglutinin from Saccharomyces cerevisiae as a membrane anchor under the control of the alcohol oxidase 1 promoter (pAOX1). Multiple cloning sites and the sequence encoding the Xpress epitope (-Asp-Leu-Tyr-Asp-Asp-Asp-Asp-Lys-) were introduced into the vector for insertion of heterologous genes and selective cleavage of target proteins. Enhanced green fluorescence protein (EGFP) was used as a model protein to check the function of this vector. The expression of EGFP on the P. pastoris surface was confirmed by confocal laser scanning microscopy. Fluorescence microscopy and western blot analysis confirmed that EGFP can be successfully cleaved from the cell surface by treating with enterokinase.
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The project was supported by a key fund from Chinese Education Ministry (2005) and a Grant from National Natural Science Foundation of China (No. 30470399).
Qingjie Wang and Lei Li contributed equally to this work.
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Wang, Q., Li, L., Chen, M. et al. Construction of a novel system for cell surface display of heterologous proteins on Pichia pastoris . Biotechnol Lett 29, 1561–1566 (2007). https://doi.org/10.1007/s10529-007-9430-6
- Enhanced green fluorescence protein
- Pichia pastoris
- Surface display