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Application of protein Com1 in immunoprotection of coxiella burnetii

A protein and protective antigen technology, applied in blood/immune system cells, antibacterial drugs, vertebrate cells, etc., can solve the problems of low reproduction amount of chicken embryos, high protection requirements, high cost, etc., to overcome preparation difficulties, Simple operation and low cost effect

Inactive Publication Date: 2013-12-04
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

Examples

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Effect test

Embodiment 1

[0032] Example 1. Preparation of antigen protein Com1

[0033] The vector pET-32a(+) was purchased from Novagen, the product catalog number is 69015. pQE30 was purchased from Qiagen Company, and the product catalog number was 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 performed with primer pairs to obtain PCR amplification products; restriction enzymes BamHI and SacI were used The PCR amplification product is digested to recover the target gene fragment; the vector pQE30 is digested with restriction enzymes BamHI and SacI to recover the large vector fragment; the target gene fragment and the large vector fragment are connected to obtain a recombinant vector; conditions for PCR amplification : Pre-denatu...

Embodiment 2

[0055] Example 2. Application of antigen protein Com1

[0056] 1. Preparation of protective antigens 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 BALB / c mouse femur bone marrow: Take BALB / c mouse femur, cut off both femoral heads with ophthalmological scissors, insert the syringe needle filled with 1ml 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 complete 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 day of culture, each well was replaced with 3ml of RPMI1640 complete medium. After 7 days of culture, the immature dendritic c...

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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 invention relates to the application of the protein Com1 in the immune protection of Coxiella peterii. Background technique [0002] Coxiella burnetii (Coxiella burnetii) is a gram-negative small bacterium that parasitizes in obligate phagocytes. The bacteria is very resistant to the external environment and has a strong infectivity to humans and animals. It can spread through aerosols and enter the body through the respiratory tract to cause infections in humans and animals and cause Q fever. [0003] Q fever is a zoonotic disease with worldwide distribution, which is widely distributed in my country. The clinical manifestations of human Q fever are diverse. Acute Q fever manifests as fever, headache, and muscle aches, often accompanied by pneumonia, hepatitis, etc.; chronic Q fever manifests as endocarditis, granulomatous hepatitis, and osteomyelitis. In recent years, Q fever meningitis, glomerulonephritis, abortion in pregnant women have also been r...

Claims

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Application Information

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Patent Type & Authority Patents(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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