Yersinia pestis protein and coding gene with immune protection function and its application

An immune protection and bacterial protein technology is applied in the application field of the preparation of plague vaccine, and achieves the effects of wide application prospect, enhanced immune protection range, and enhanced immune protection effect.

Inactive Publication Date: 2012-02-08
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on cellular immunity after plague infection. Recent studies have found that the T cell immunity produced by Y. May play a role (Philipovskiy AV, Smiley ST, Vaccination with live Yersinia pestis primesCD4 and CD8 T cells that synergistically protect against lethal pulmonary Y. pestis infection. Infect Immun 2007, 75(2): 878-885)

Method used

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  • Yersinia pestis protein and coding gene with immune protection function and its application
  • Yersinia pestis protein and coding gene with immune protection function and its application
  • Yersinia pestis protein and coding gene with immune protection function and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Example 1. Screening of Yersinia pestis protein with immune protection function

[0057] The following method is used to screen the Yersinia pestis protein with immune protection function, and the specific process includes the following steps:

[0058] 1. Selection of candidate genes and cloning and expression in Escherichia coli

[0059] Using PSORTb, SignalP and ProtcompB software to analyze the cellular location of 4016 genes encoding proteins on the chromosome of Yersinia pestis CO92 strain (the first sequenced strain of Yersinia pestis), the results showed that 1006 genes encoded proteins were membrane-associated , secreted proteins; the transmembrane domains of these proteins were analyzed by TopPred software. Since proteins with multiple transmembrane domains are difficult to express in vitro, all proteins with more than 3 transmembrane domains were removed. Results 430 genes encoded proteins with less than 4 transmembrane regions; among these 430 genes, combine...

Embodiment 2

[0070] Example 2. Prokaryotic expression of Yersinia pestis protein with immune protection function and purification of expression product

[0071] The following prokaryotic expression method is used to obtain the Yersinia pestis protein with cellular immune protection function. The specific method is: using site-specific recombination and Gateway cloning technology, the corresponding gene coding sequence of the Yersinia pestis protein with immune protection function (YPO0606, YPO0612, YPO1914, YPO3119, YPO3047, YPO0420, YPO1377 and YPO3720) were cloned into the expression vector pDEST17 (invitrogen company), and the prokaryotic expression vectors of Yersinia pestis protein with immune protection function were obtained (respectively named as pDEST17-YPO0606, pDEST17-YPO0612, pDEST17 -YPO1914, pDEST17-YPO3119, pDEST17-YPO3047, pDEST17-YPO0420, pDEST17-YPO1377 and pDEST17-YPO3720), the Yersinia pestis protein YPCD1.05c coding sequence with immune protection function was cloned in...

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Abstract

The invention discloses a plague bacteria protein with cellular immune protection function, an encoding gene and an application. The purpose is to provide the protein and the encoding gene which can stimulate T cell immune reaction during plague bacteria infection and immune processes and can generate cellular immune protection function, as well as the application in the preparation of plague vaccines. The experiments prove that the protein of the invention can stimulate T cell immune reaction during the infection and the immune processes of plague bacteria and can generate immune protection function. The protein or gene of the invention can be taken as an active ingredient for preparing the plague vaccines, and can enhance the immune protection effect and the immune protection region of the existing plague vaccines through mutual immunization or the combined immunization with the existing F1 and LcrV antigen or gene with immune protection function, or the preparation of a recombinant fusion multivalent subunit vaccine or DNA vaccine at the same time. The invention provides a new means for searching the protein of the plague bacteria with cellular immune protection function, which can play the important role in the prevention and the treatment of plague and have broad application prospect.

Description

technical field [0001] The present invention relates to proteins and corresponding coding genes and their applications, in particular to proteins and corresponding coding genes capable of stimulating T cell immune responses and producing immune protection functions during Yersinia pestis infection and immunization and their applications in the preparation of plague vaccines . Background technique [0002] Plague is a class A infectious disease caused by Yersinia pestis, which has caused three worldwide pandemics in history. In 2000, the World Health Organization listed plague as a re-emerging infectious disease. The emergence of multidrug-resistant plague strains and the possibility that Y. pestis could be used as biological weapons and bioterrorism brought difficulties to the prevention and treatment of Y. pestis. Effective vaccination can block the spread of plague and achieve the ultimate goal of eradicating plague. However, so far, there is no plague vaccine that can b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/24C12N15/31C12N15/63C12N15/70C12N5/10C12N1/15C12N1/19C12N1/21A61K39/02A61K31/7088A61K48/00A61P31/04
CPCY02A50/30
Inventor 李蓓郭景玉王效义杨瑞馥
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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