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

一种噬菌体、工程化的技术，应用在文库、微生物库、组合化学等方向，能够解决多肽尚未见报道等问题

Inactive Publication Date: 2011-02-02
JANSSEN BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous reports on pIX phage display described fusions in the context of phagemid vectors; display of polypeptides on pIX from hybrid vectors or phage vectors has not been reported

Method used

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  • 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
  • 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
  • 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

Examples

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example 1

[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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Abstract

The invention relates to a compositions and methods for generating and using pIX phage display libraries for producing non-antibody peptide or protein proteins or peptides using engineered hybrid phage vectors derived from pIX of M 13 phage.

Description

technical field [0001] The present invention relates to compositions and methods for generating a pIX phage display library using engineered hybrid phage vectors derived from pIX of M13 bacteriophage and using the library to generate one or more non-antibody peptides or proteins. Background technique [0002] Phage display is a well-established tool for affinity-based peptide selection. (In a typical phage display selection, a library of polypeptides is engineered to be fused to the terminus of one of the coat proteins of the filamentous bacteriophage M13. The phage particle provides a physical association between each polypeptide member of the library and the gene it encodes. For library members that bind to the desired target molecule, perform affinity selection on the phage library, that is, panning.The library is mixed with the target molecule, unbound phage particles are washed away, and the remaining phage are eluted and cultured in E. coli cells And amplify. [0003...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/00
CPCC12N15/1037C40B40/02
Inventor B·王L·尹K·奥奈尔
Owner JANSSEN BIOTECH INC
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