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Antigen simulating epipeptide of C type para chicken blood phili bacillus and its use

A technology of haemophilus gallinaceus and mimic epitopes, which is applied in the field of differential diagnosis of chicken infectious rhinitis and vaccine development, can solve problems affecting the growth of laying hens, achieve good economic benefits and market prospects, and be easy to use The effect of the operation

Inactive Publication Date: 2007-03-14
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is necessary to use a large number of inactivated bacteria and adjuvants of chemical materials. Hpg is a Gram-negative bacterium that contains endotoxin. Mass inoculation will affect the egg production of laying hens and the growth of rearing chickens

Method used

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  • Antigen simulating epipeptide of C type para chicken blood phili bacillus and its use
  • Antigen simulating epipeptide of C type para chicken blood phili bacillus and its use
  • Antigen simulating epipeptide of C type para chicken blood phili bacillus and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Screening and sequence analysis of Haemophilus paragallinarum antigen mimotope type C

[0029] Experimental Materials:

[0030] 1. Phage Random Peptide Library Kit

[0031] Linear 12-peptide library displayed on phage surface capsid protein III (Ph.D.-12 TM Phage Peptide Library PeptideLibrary Kit) kit was purchased from New England Biolabs, USA. The titer of the peptide library was 4×10 12 pfu / ml, random diversity 1.9×10 9 ; The genotype of the recipient strain E.coli ER2537 is

[0032] F'lac q Δ(lacZ)M15proA + B + fhuA2supE thiΔ(lac-proAB)Δ(hsdMS-mcrB)

[0033] 5(r k - m k - McrBC - ). DNA automatic sequencing primer (-96gIII sequencing primer):

[0034] 5'-CCCTCATAGTTAGCGTAACG-3'

[0035] 2. One anti-Haemophilus paragallinarum monoclonal antibody HPG-C-44; one anti-Haemophilus paragallinarum monoclonal antibody HPG-A-48; preserved by the Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Scien...

Embodiment 2

[0113] Example 2 Application of Mimotope Fusion Peptides in Differential Diagnosis

[0114] Experimental Materials

[0115] Host bacteria: GI826(F - ,lacI q ,ampC::P trp cI, ΔfliC, ΔmotB, eda::Tn10), products of Invitrogen;

[0116] Plasmid: pFliTrx Invitrogen;

[0117] main reagent

[0118] Restriction endonuclease: ApaI, product of Xho I TaKaRa company;

[0119] T 4 DNA ligase: TaKaRa company product;

[0120] Taq polymerase: product of TaKaRa Company.

[0121] Related media

[0122] 10XM9 Salt: Na 2 HPO 4 60g, KH 2 PO 4 30g, NaCl 5g, NH 4 Dissolve 10g of Cl in 900ml of deionized water, and adjust the pH to 7.4 with 10M NaOH;

[0123] IMC medium: containing 1×M9 salt solution, 0.2% casein acid hydrolyzate, 0.5% glucose, 1mmol / L MgCl 2 ;

[0124] RM medium: 1XM9 salts, 2% tyrosine, 1% glycerol, 1mMMgCl 2 ;

[0125] SOC medium: tryptone 20g, yeast extract 5g, NaCl 0.5g, 2.5mmol / L KCl, 10mmol / LMgCl 2 , 20mmol / L glucose.

[0126] Rabbit anti-chicken IgG-HR...

Embodiment 3

[0163] Example 3: Application of Mimotope Fusion Peptides in Vaccines

[0164] 3.1 Experimental animals and materials

[0165] Fifty-six 45-day-old non-immune chickens were purchased from Beijing Institute of Experimental Animals; the induced recombinant strain F44; the induced host strain F0 with an empty vector; HPG-C live bacteria, etc., were preserved in our laboratory; SPF chicken anti-HPG-C antiserum, HPG oil emulsion inactivated vaccine, etc., are preserved in our laboratory.

[0166] 3.2 Induced expression of recombinant bacteria displaying mimotopes

[0167] Inoculate the bacterial liquid of the recombinant bacteria (named F44) correctly identified by sequencing into RM medium (the ampicillin concentration is 100 μg / μl) at a ratio of 1:100, and shake culture at 30°C for 15 hours (about 2.0-3.0); Inoculate 40 μl of overnight bacteria into 2ml IMC medium, induce with tryptophan (concentration: 100 μg / ml), culture with shaking at 37°C for 6 hours, collect bacteria, and...

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Abstract

The invention relates to the antigen imitated epitope peptide of Haemophilus paragallinarum serovar C in China and its application, wherein its amino acid sequence is represented by the general formula of YX1PX2QWWX3X4X5X6X7, wherein Y represents tyrosine residue, P represents proline residue, Q represents glutamine residue, W represents tryptophan residue, X1-X7 represents any amino acids. The peptide can be used for providing an easily-manipulated serological type differential diagnosis method for chicken infectious rhinitis.

Description

technical field [0001] The present invention relates to short peptides simulating antigenic epitopes of Haemophilus paragallinarum, in particular to a group of short peptides capable of simulating the serotype epitopes of Haemophilus paragallinarum type C, and the use of the group of short peptides in preparing chicken infectious Differential diagnosis of rhinitis and application in vaccine development. Background technique [0002] Haemophilus paragallinarum (Hpg) is a pathogenic bacterium that causes infectious rhinitis in chickens. It can be divided into three serotypes: A, B, and C. Types A and C are common in my country now. Chicken infectious rhinitis is an acute respiratory infectious disease. Its main hazards are the decrease in egg production of laying hens and poor growth of reared chickens. It has been reported in the literature that infectious rhinitis caused by Hpg can reduce egg production by 40%, causing serious economic losses. [0003] The identification ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08A61K39/07A61P31/04G01N33/53G01N33/569
Inventor 张培君王宏俊柳川高亚萍龚玉梅孙惠玲
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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