Application of protein to preparation of medicine for preventing trueperella pyogenes infection

A technology of cryptobacillus pyogenes and protein, applied in application, antibacterial drugs, bacterial peptides, etc., can solve the problem of lack of new protective antigens

Active Publication Date: 2020-01-03
CHONGQING ACAD OF ANIMAL SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] The identified virulence factors of C. pyogenes include hemolysin (Pyolysin, PLO), adhesion protein collagen-binding protein (collage-binding protein, CbpA), neuraminidases (Nan) NanH and NanP, etc., and PLO is the main one. protective antigens, no new protective antigens have been discovered so far

Method used

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  • Application of protein to preparation of medicine for preventing trueperella pyogenes infection
  • Application of protein to preparation of medicine for preventing trueperella pyogenes infection
  • Application of protein to preparation of medicine for preventing trueperella pyogenes infection

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Experimental program
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Embodiment

[0032] 1. Analysis of immunoreactive membrane proteins

[0033] 1. Antiserum preparation of C. pyogenes

[0034] The strain is the Chongqing isolate of Cryptobacterium pyogenes in the patent application number CN201410072433.8 (biological deposit number: CCTCC NO: M2014037, (GenBank NO: CP012649).

[0035]A single colony of C. pyogenes was picked to inoculate TSB medium, and cultured at 37°C for 48 hours. Add 0.2% formaldehyde solution to the culture, inactivate it at 37°C for 24 hours, and shake it 5 times during the period. Mix and emulsify the inactivated culture with Freund's adjuvant to prepare the inactivated antigen. Leg intramuscular injection, 0.2mL / mouse, immunized once every 2 weeks, immunized 3 times in total. Fifteen days after the third immunization, the blood of the mice was collected by enucleating the eyeballs, and centrifuged to collect the antiserum of C. pyogenes after blood coagulation.

[0036] 2. Membrane protein separation and mass spectrometry anal...

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Abstract

The invention belongs to the field of a veterinary biological product, and discloses an antigen protein of trueperella pyogenes, preparation of the antigen protein, application of the antigen proteinto a vaccine, and an antibody detection method. A gene of a highly conservative trueperella pyogenes Fe ABC (ATP Binding Cassette) transporter SBP (Substrate-Binding Protein) is cloned to a pGEX-4T-1plasmid; a BL21(DE3) recombinant strain is built; IPTG is used for induction expression of the recombinant protein SBP (GST-SBP); the soluble expression of the target protein is realized; and the recombinant protein is subjected to affinity purification by a Gluthathione-Sepharose 4B chromatography method. The recombinant SBP (GST-SBP) protein provided by the invention has high reactionogenicity and immunogenicity, and can react with the blood serum of trueperella pyogenes immunized or infected animals (mice and goats); the animals (mice and goats) can be stimulated to generate high-level specific antibodies; and the immune protective rate of the mice is 90 percent. The novel antigen protein is provided for the preparation of medicine for preventing the trueperella pyogenes infection.

Description

technical field [0001] The invention belongs to veterinary biological products, in particular to an antigen protein for preparing and preventing infection of Cryptobacter pyogenes. Background technique [0002] The goat industry has developed rapidly in the Three Gorges reservoir area in recent years, and has become one of the important industries that help farmers get rid of poverty and become rich and the healthy development of rural areas. However, the impact of epidemic diseases on sheep farming in the region is becoming more and more apparent. Among them, suppurative pneumonia is more harmful. Because of its long latent infection period and no obvious clinical symptoms, suppurative lesions cause irreversible damage to lung tissue, which seriously affects the survival rate of lambs. Trueperella pyogenes is one of the important pathogens of goat suppurative pneumonia (Zhang Suhui et al., 2015; Fu Lizhi et al., 2018; Feng Feng, 2010.). Cryptobacterium pyogenes is a Gram-p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/195C12N15/31C12N15/70G01N33/68A61K39/02A61P31/04
CPCC07K14/195C12N15/70G01N33/68A61K39/0208A61P31/04G01N2333/195A61K2039/552Y02A50/30
Inventor 沈克飞张素辉闫志强徐登峰许国洋付利芝
Owner CHONGQING ACAD OF ANIMAL SCI
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