Application of mycoplasma bovis secretory protein MbovP280

A technology of Mycoplasma bovis and secreted protein, applied in the field of animal infectious disease prevention and control, can solve problems such as unclear

Active Publication Date: 2020-10-30
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the mechanism of mycoplasma bovis-induced host macrophage apoptosis is still unclear, and the signific

Method used

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  • Application of mycoplasma bovis secretory protein MbovP280
  • Application of mycoplasma bovis secretory protein MbovP280
  • Application of mycoplasma bovis secretory protein MbovP280

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: Expression of Mycoplasma bovis MbovP280 protein

[0026] 1. Cloning and expression of Mycoplasma bovis Mbov_0280 gene

[0027] Because E. coli is to the preference of codon, the codon UGA of tryptophan of encoding tryptophan in E. coli is used as terminator in E. coli in the present invention, therefore, when expressing M. bovis gene with E. coli, need carry out mycoplasma gene Mutation, the codon UGA is mutated to the codon UGG that can express tryptophan in E. coli.

[0028] The applicant isolated the local isolate of Mycoplasma bovis in June, 2008 from the lung tissue of a yellow cattle that fell ill in a cattle farm in Yingcheng City, Hubei Province, and named it Mycoplasma bovis HB0801, Mycoplasma bovis HB0801. The Mbov_0280 gene in GenBank accession number CP002058) was used as a template, and the nucleotide sequence of the gene was optimized according to the codon preference of Escherichia coli, and the optimized sequence was sent to the company fo...

Embodiment 2

[0043] Example 2: Functional verification of Mycoplasma bovis rMbovP280

[0044] The cell viability after 24h stimulation with different proteins was detected by CCK-8 method. 1 x 10 per well 4 Seed BoMac or RAW cells in a 96-well plate at 37°C, 5% CO 2 Culture until adherent, set blank wells (no cells) and control wells (only cells), set 3 replicate wells for each group; place at 37°C, 5% CO 2 Incubate overnight and observe under an inverted microscope. Take out the cell plate, add 4 μg rMbovP280 protein to each well, 37°C, 5% CO2 static co-cultivation for 24 hours, add 10 μL CCK-8 (Japanese colleagues) to each well, incubate at 37°C, 5% CO2 for 1 hour, measure the absorbance of each well at 450 nm value, calculate the cell viability of each well according to the following formula, that is, cell viability%=(OD 加蛋白细胞 -OD 空白 ) / (OD 无蛋白对照 -OD 空白 )×100%. The experimental results showed that the experimental results showed that 4μg MbovP280 protein stimulated 1×10 4 RAW264...

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Abstract

The invention discloses application of mycoplasma bovis secretory protein MbovP280 in preparation of mycoplasma bovis diagnostic reagents, drugs or vaccines, and belongs to the technical field of animal infectious disease prevention and treatment. The mycoplasma bovis secretory protein MbovP280 has an amino acid sequence shown as SEQ ID NO:2; and tests prove that the mycoplasma bovis secretory protein MbovP280 has the capacity of inducing host macrophage activity reduction and apoptosis, and has important potential in the aspects of research and development of new drugs and vaccines.

Description

technical field [0001] The invention belongs to the technical field of animal infectious disease prevention and control, and specifically relates to the application of a mycoplasma bovis secretory protein MbovP280, which has the ability to induce host macrophage viability decline and apoptosis. Background technique [0002] Mycoplasma bovis (M.bovis) belongs to the class Mollicutes, the order Mycoplasmatles, the family Mycoplasma, and the genus Mycoplasma. It is a minimal pathogenic microorganism that can replicate in vitro. Mycoplasma bovis can infect cattle of any age, and calves aged 2 to 6 months are most susceptible (Stipkovits et al 2000). The clinical symptoms after the onset of beef cattle are mainly pneumonia and arthritis, and the main manifestations of dairy cattle are mastitis. After Mycoplasma bovis infects calves, it is often difficult to cure, and it becomes a chronic disease after counterbalancing with the body's immune system. When mycoplasma bovis occurs, ...

Claims

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Application Information

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IPC IPC(8): G01N33/569A61K39/02A61P31/04C07K14/30C12N15/31C12N15/70
CPCA61K39/0241A61P31/04C07K14/30C12N15/70C12N2800/22G01N33/56933G01N2333/30
Inventor 郭爱珍赵刚伊赫萨努拉·希拉尼陈颖钰胡长敏陈曦陈建国
Owner HUAZHONG AGRI UNIV
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