Method for preparing phase granule for displaying polypeptide

A technology of phagemids and particles, which is applied in the field of preparing phagemid particles displaying polypeptides, which can solve the problems of high production cost and low yield of helper phages

Inactive Publication Date: 2006-12-27
SHANGHAI INST OF ONCOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, we found in our research that if you want to directly use helper phages that express some relatively large pro

Method used

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  • Method for preparing phase granule for displaying polypeptide
  • Method for preparing phase granule for displaying polypeptide
  • Method for preparing phase granule for displaying polypeptide

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Embodiment Construction

[0036] The present invention provides a method for preparing phagemid particles displaying polypeptides. The method adopts a genomic plasmid encoding a helper phage of a fusion protein of exogenous polypeptide and pIII coat protein to co-transform with phagemid or successively transform F factor positive strains , Thereby obtaining phagemid particles displaying exogenous polypeptides. The phagemid particles obtained by this method have the characteristics of high display level of exogenous polypeptides, large capacity of accommodating exogenous target genes, and small virus particles. This method can be used to prepare phage gene transfer vectors and prepare antigen-displaying vectors as antigens.

[0037] The method can be to transform the helper phage genome plasmid and phagemid particles into F factor positive strains at the same time; it can also be to transform the helper phage genome plasmid into F factor positive strains, and then transform the phagemid to contain the helpe...

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Abstract

This invention involves the phagemid in molecule biology field and medicine field. It is a preparation method of display peptide's phagemid. This method adopts the gene group plasmid of assistant phage fusion protein of coding foreign peptide and pIII coat protein, to co-transformation or successive-transformation F factor positive Isolates together with phagemid, so to obtain phagemid that display foreign peptide. The phagemid prepared by this method has high foreign peptide display level, and it is not necessarily prepare assistant phage so to simplified the process of preparation of assistant phage.

Description

Technical field: [0001] The invention relates to the preparation of phagemid particles in the fields of molecular biology and medicine, in particular to a method for preparing phagemid particles that display polypeptides. Background technique: [0002] Wild-type filamentous phage (filamentous phage) contains a single-stranded closed-loop DNA molecule of approximately 6.4 kb. The virus particle has a diameter of about 6.5 nm and a length of about 900 nm. The outer shell is mainly composed of the main coat protein (pVIII, 2700 copies). There are only 3 to 5 copies of the minor coat protein (pIII). Virus particles can only infect bacterial strains with sexual cilia. When the virus passes through the sexual cilia, the main outer coat protein (pVIII) falls off, and the viral DNA enters the cell and is transformed into double-stranded DNA under the action of intracellular enzymes. [0003] So far, pIII, pVIII, and pVI have all been used to display peptide ligands, protein ligands, anti...

Claims

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Application Information

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IPC IPC(8): C12N15/86C12N7/01C12N15/62C07K19/00
Inventor 李宗海顾健人杨胜利张捷石必枝
Owner SHANGHAI INST OF ONCOLOGY
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