Engineered hybird phage vectors for the design and the generation of a human non-antibody peptide or protein phage library via fusion to pix of m13 phage
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[0057] Example 1: Construction of an exemplary engineered phage vector
[0058] Phage construction. M13-99 and M13-99L contain the recombinant pIX gene inserted into the phage genome M13KE (a derivative of M13mp19). This recombination region was inserted into the lacZα region of M13mp19, which is located in the intergenic region of the phage genome, so that the lac promoter can drive the transcription of the recombinant gene IX fusion. insertion sequence ( figure 1 ) contains the Shine-Delgarno sequence (ribosome binding site), the signal sequence for pectate lyase B (pelB), the double BbsI restriction enzyme recognition site for subsequent cloning, the pIX coding region, and the trpA transcription terminator. FLAG tag peptide DYKDDDDK (SEQ ID NO: 2) and five amino acid linker (M13-99: GGTKT (SEQ ID NO: 7)) or nine amino acid linker (M13-99L: SGGSGGTKT (SEQ ID NO: 8)) contained in pelB and between gene IX.
[0059] Four additional peptides (Table 1 ) of various lengths a...
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