Document Detail

Construction of a novel Pichia pastoris cell-surface display system based on the cell wall protein Pir1.
MedLine Citation:
PMID:  18188644     Owner:  NLM     Status:  MEDLINE    
A novel system based on Pir1 from Saccharomyces cerevisiae was developed for cell-surface display of heterologous proteins in Pichia pastoris with the alpha-factor secretion signal sequence. As a model protein, enhanced green fluorescence protein (EGFP) was fused to the N-terminal of the mature peptide of Pir1 (Pir1-a). The expression of fusion protein EGFP-Pir1-a was irregular throughout the P. pastoris cell surface per detection by confocal laser scanning microscopy. A truncated sequence containing only the internal repetitive sequences of Pir1-a (Pir1-b) was used as a new anchor protein in further study. The fusion protein EGFP-Pir1-b was expressed uniformly on the cell surface. The fluorescence intensity of the whole yeast was measured by spectrofluorometer. Western blot confirmed that the fusion proteins were released from cell walls after mild alkaline treatment. The results indicate that a Pir1-based system can express proteins on the surface of P. pastoris and that the fusion proteins do not affect the manner in which Pir1 attaches to the cell wall. The repetitive sequences of Pir1 are required for cell wall retention, and the C-terminal sequence contributes to the irregular distribution of fusion proteins in P. pastoris.
Qingjie Wang; Lei Li; Min Chen; Qingsheng Qi; Peng George Wang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-01-09
Journal Detail:
Title:  Current microbiology     Volume:  56     ISSN:  0343-8651     ISO Abbreviation:  Curr. Microbiol.     Publication Date:  2008 Apr 
Date Detail:
Created Date:  2008-02-28     Completed Date:  2008-08-15     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  7808448     Medline TA:  Curr Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  352-7     Citation Subset:  IM    
Life Science School, Shandong University, Jinan, 250100, People's Republic of China.
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MeSH Terms
Blotting, Western
Cell Wall / genetics,  metabolism
Fungal Proteins / genetics*,  metabolism*
Genes, Reporter
Genetic Vectors
Green Fluorescent Proteins / genetics,  metabolism
Microscopy, Confocal
Peptides / genetics
Pichia / genetics*,  metabolism*
Protein Sorting Signals
Recombinant Fusion Proteins / genetics*,  metabolism*
Repetitive Sequences, Amino Acid / genetics
Saccharomyces cerevisiae Proteins
Spectrometry, Fluorescence
Reg. No./Substance:
0/Fungal Proteins; 0/Glycoproteins; 0/PIR1 protein, S cerevisiae; 0/Peptides; 0/Protein Sorting Signals; 0/Recombinant Fusion Proteins; 0/Saccharomyces cerevisiae Proteins; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins; 61194-02-3/mating factor

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