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Phage vector for dual-epitope display and construction method thereof

A phage, double epitope technology, applied in the field of genetic engineering, can solve the problems of unable to display fragments, affecting phage assembly, etc., to achieve the effect of enriching species

Inactive Publication Date: 2015-11-04
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The pⅧ protein is the main coat protein of filamentous phage, located on the back of the phage, with about 2,700 copies. It can display a large number of foreign polypeptides, but it cannot display too long fragments, otherwise it will affect the assembly of the phage

Method used

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  • Phage vector for dual-epitope display and construction method thereof
  • Phage vector for dual-epitope display and construction method thereof
  • Phage vector for dual-epitope display and construction method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1. Construction of phage vectors for double-epitope display

[0040] In this example, the gene encoding the recombinant pVIII protein (denoted as rgVIII gene) on the phage vector f88-4 is cut out and inserted into the phage vector fUSE55, thereby constructing a phage vector for double-epitope display.

[0041]Design ideas: On the phage vector f88-4, find a specific restriction enzyme site at both ends of the rgⅧ gene. These two restriction enzyme sites must not only cut the rgⅧ gene, but also It is guaranteed that there is only a corresponding single site in the phage vector fUSE55, and other structural or functional elements of the phage vector fUSE55 will not be cut off. Through analysis, it was found that the genome of phage f88-4 contained many different restriction enzyme cutting sites, but only Msc Ⅰ and Sac Ⅱ restriction enzymes met the above conditions. Therefore, the inventors of the present invention decided to use these two restriction endonucleases ...

Embodiment 2

[0070] Examples of the application of the phage vectors for double-epitope display constructed in Example 2 and Example 1

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Abstract

The invention disclsoes a phage vector for dual-epitope display and a construction method thereof. The phage vector for dual-epitope display is a filobactivirus-originated annular vector used for displaying coding gene of pIII protein and pVIII protein of exogenous peptides. By integrating a phage pIII display system and a pVIII display system for the first time, a phage pIII and pVIII dual-epitope display system is formed. Different exogenous peptides can be displayed on pIII protein and pVIII protein of a same phage. Therefore, types of the phage display system are enriched, and new tools and methods are provided for biological and medical application of the system.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and relates to a phage carrier used for double-epitope display and a construction method thereof. Background technique [0002] Phage display technology refers to the use of phage as a carrier to insert foreign genes into the phage genome and express them along with the expression of the phage genome. When the phage self-assembles, the protein or polypeptide expressed by the foreign gene is used as the coat protein. A part is displayed on the surface of phage in the form of fusion protein. [0003] Since Gorge P. Smith first proposed phage display in 1985, phage display technology has been widely used, and a variety of phage display systems have been formed, including pIII display system, pVIII display system and pVI display system. The pIII protein is the secondary coat protein of the virus, which is located at the tail of the virus particle. Since the pIII protein has a small number of copie...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/66C12N7/01
Inventor 王丽鞠志刚高翔
Owner NORTHEAST NORMAL UNIVERSITY
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