Use of the arabinose p(bad) promoter for tightly regulated display of proteins on bacteriophage.
Phage display is a widely used method to optimize the binding characteristics of protein-ligand interactions. In addition, it has been used to clone genes from genomic and cDNA libraries based on their ligand-binding characteristics. One difficulty often encountered when expressing heterologous proteins by phage display is the toxicity of the protein on the Escherichia coli host. Previous studies have shown that heterologous protein expression can be tightly controlled using plasmids with the P(BAD) promoter of the arabinose operon of E. coli, and the araC gene, which is both a positive and negative regulator of the promoter. We constructed a set of phage display vectors that utilize the P(BAD) promoter to control the expression of proteins on the surface of the M13 bacteriophage. These vectors exhibit tightly controlled expression of proteins on the surface of the phage. In addition, the amount of protein displayed on the phage is modulated by the amount of arabinose present in the growth medium during phage propagation. This may be useful for altering the stringency of binding enrichment during phage display.
Huang
W
W
Department of Molecular Virology & Microbiology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA.
McKevitt
M
M
Palzkill
T
T
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D051777:AraC Transcription Factor; D001089:Arabinose / Q000378:metabolism*; D001426:Bacterial Proteins; D017104:Bacteriophage M13 / Q000235:genetics*; D001483:Base Sequence; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D005786:Gene Expression Regulation; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics; D015698:Genomic Library; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics; D014157:Transcription Factors; D014771:Virion / Q000187:drug effects / Q000235:genetics; D001618:beta-Lactamases / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D051777:AraC Transcription Factor; C497371:AraC protein, E coli; D001426:Bacterial Proteins; D004274:DNA, Recombinant; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D001089:Arabinose; D001618:beta-Lactamases