Figure 1
From: Direct selection and phage display of a Gram-positive secretome
Phage display vector for selective secretome display. C-gIII, carboxy-terminal domain of gIII; CmR, chloramphenicol resistance cassette; colE1 ori, the colE1 plasmid origin of replication; ppsp, phage shock protein promoter; MCS, multiple cloning site; RBS, ribosomal binding site; C-myc, a common peptide tag followed by a single amber stop codon; f1 ori, the f1 phage origin of replication for generation of ssDNA for packaging into the phagemid particles. The stop codon is read as glutamic acid in the host strain TG1 (supE) used in the library construction and screening, allowing read-through into the in-frame gIII-coding sequence and display on the phage. Expression of the soluble secretome proteins tagged with the C-myc peptide tag (without pIII moiety) can be achieved by using a suppressor-negative E. coli host strain.