Displaying non-natural, functional molecules on yeast surfaces via biotin-streptavidin interaction.
Here we expand the yeast cell surface display system to display non-natural, functional molecules. The short biotin acceptor peptide (BAP) sequence of biotin ligase from E. coli (BirA) was genetically introduced to the N-terminus of the anchor protein, Flo428. Through co-expression of BAP-fused Flo428 with BirA, biotinylated BAP could be displayed on the yeast cell surface. Subsequent addition of streptavidin-FITC resulted in the display of streptavidin-FITC, and, the display of biotin-FITC was successful using streptavidin as a linker. Our strategy provides a powerful tool for displaying functional molecules on yeast cell surfaces.
Kobe University, Rokkodai-cho, Nada, Japan.
Tsutomu
T
Tanaka
Shinsuke
S
Masunari
Jun
J
Ishii
Kanako
K
Wakamura
Maiko
M
Segawa
Hideki
H
Fukuda
Akihiko
A
Kondo
D001710:Biotin / Q000378:metabolism*; D019921:Biotinylation; D015153:Blotting, Western; D019731:Carbon-Nitrogen Ligases / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D019809:Streptavidin / Q000378:metabolism*; D015003:Yeasts / Q000166:cytology / Q000378:metabolism*
D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; D012097:Repressor Proteins; D001710:Biotin; D019809:Streptavidin; D019731:Carbon-Nitrogen Ligases; C058125:birA protein, E coli