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Application of protein Com1 in immunoprotection of coxiella burnetii

A technology of coxe bodies and proteins, which is applied in blood/immune system cells, antibacterial drugs, vertebrate cells, etc., can solve the problems of low reproductive capacity of chicken embryos, high protection requirements, and high cost, and achieve the goal of overcoming Difficult to prepare, easy to operate and low cost

Inactive Publication Date: 2011-08-03
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

Both inactivated vaccines and chemically extracted subunit vaccines require a large number of chicken embryos to multiply Coxiella beinii for preparation, but the number of chicken embryos is low, and the protection requirements for extracting and purifying Coxiella beinii are high, the process is complicated, and the cost is expensive , so it is difficult to prepare and promote the use of large quantities

Method used

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  • Application of protein Com1 in immunoprotection of coxiella burnetii
  • Application of protein Com1 in immunoprotection of coxiella burnetii
  • Application of protein Com1 in immunoprotection of coxiella burnetii

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Embodiment 1

[0032] Embodiment 1, the preparation of antigenic protein Com1

[0033] The vector pET-32a(+) was purchased from Novagen, the product catalog number is 69015. pQE30 was purchased from Qiagen Company, the product catalog number is 33203.

[0034] Escherichia coli BL21 was purchased from Novagen, the product catalog number is 69450. Escherichia coli M15 was purchased from Novagen, the product catalog number is 34210.

[0035] Preparation of the coding gene of protein Com1: using the genomic DNA of Coxiella burnetii as a template, PCR amplification was carried out with primer pairs to obtain the PCR amplification product; restriction enzymes BamHI and SacI were used Digest the PCR amplification product and recover the target gene fragment; digest the vector pQE30 with restriction enzymes BamHI and SacI to recover the large vector fragment; connect the target gene fragment and the large vector fragment to obtain the recombinant vector; PCR amplification conditions : Pre-denatur...

Embodiment 2

[0055] Embodiment 2, the application of antigenic protein Com1

[0056] 1. Preparation of protective antigen against Q fever

[0057] 1. Preparation of immature dendritic cells

[0058] BALB / c mice were purchased from the Experimental Animal Center of the Academy of Military Medical Sciences.

[0059] Isolate bone marrow cells from the femoral bone marrow of BALB / c mice: Take the femur of BALB / c mouse, cut off the femoral heads on both sides with ophthalmic scissors, insert the syringe needle filled with 1ml of PBS buffer into the bone cavity, and slowly push the syringe to rinse , Flush a single bone marrow cell suspension from the bone cavity.

[0060] Adjust the number of bone marrow cells to 1×10 with RPMI1640 full medium 6 / ml, and transferred to a six-well culture plate (9.6×6cm 2 ), cultured at a temperature of 37°C and a carbon dioxide volume concentration of 5%, on the 3rd and 5th days of culture, each well was replaced with 3ml of RPMI1640 full medium, and cultur...

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Abstract

The invention discloses the application of a protein Com1 in the immunoprotection of coxiella burnetii, which is the application of the protein as shown in SEQ ID NO:1 in the preparation of a Q hot vaccine. The application proves that the recombinant protein Com1 of the coxiella burnetii can serve as an antigen protein, can stimulate an organism to produce an immune response to fight against the attack on virus strains of the coxiella burnetii, and is a protective antigen which has the functions of the Q hot vaccine. The protein Com1 is prepared through a gene engineering method, and the protein Com1 has the advantages of being simple and convenient to operate, needing low cost, and having high yield. As the protein Com1 serves as the Q hot vaccine, the defect of the difficult preparation of vaccines in the prior art is overcome.

Description

technical field [0001] The present invention relates to the application of protein Com1 in the immune protection against Coxella beinerii. Background technique [0002] Coxiella burnetii is a small Gram-negative bacillus parasitic in obligate phagocytic cells. The bacterium has strong resistance to the external environment and has strong infectivity to humans and animals. It can enter the body through aerosol diffusion through the respiratory tract, causing infection in humans and animals and Q fever. [0003] Q fever is a zoonotic disease with worldwide distribution and is widely distributed in my country. The clinical manifestations of Q fever in humans are diverse. Acute Q fever manifests as fever, headache, muscle aches, often accompanied by pneumonia, hepatitis, etc.; chronic Q fever manifests as endocarditis, granulomatous hepatitis, osteomyelitis, etc. In recent years, Q fever meningitis, glomerulonephritis, and abortion in pregnant women have also been reported. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/02C12N5/0784A61P31/04
Inventor 熊小路温博海王锡乐
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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