Recombinant pasteurella multocida toxin protein and application thereof

A pasteurella, recombinant protein technology, applied in antibacterial immunoglobulin, application, complement protein and other directions, can solve the problems of reduced feed utilization efficiency, pig jaundice, high pollution rate, etc., to improve immunity and efficiency, The effect of increasing the specific immune response

Active Publication Date: 2014-09-24
维克香港贸易有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Inoculation of PMT toxin into four-week-old piglets by means of muscle, intraperitoneal and nasal inoculation, etc., can cause turbinate and interosseous atrophy of lesions, in addition, it will also affect the general bone development and cause growth retardation, even at high doses of inoculation, Can cause liver damage leading to jaundice and death in pigs
[0004] Porcine atrophic rhinitis (AR) spreads all over the pig raising areas in the world, and the affected pigs grow slowly and the feed utilization efficiency decreases
Although the mortality rate is not high when infected alone, the pollution rate is very high, and it is easy to induce other complications or infection of pathogens, resulting in high mortality and increased production costs

Method used

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  • Recombinant pasteurella multocida toxin protein and application thereof
  • Recombinant pasteurella multocida toxin protein and application thereof
  • Recombinant pasteurella multocida toxin protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1 Construction of Pasteurella septica toxin recombinant protein (rPMT)

[0060] 1. Design of the amino acid sequence of Pasteurella septica toxin recombinant protein 1 (rPMT1)

[0061] First, using the full-length amino acid sequence of Pasteurella septica toxin protein (PMT) (as shown in SEQ ID NO: 1) as a template, using an epitope prediction website (such as, but not limited to, BCPred( http: / / ailab.cs.iastate.edu / bcpreds / )) Predicted epitopes of Pasteurella septica toxin protein (PMT).

[0062] The epitopes predicted by Epi are shown in Table 1 (SEQ ID NOs:20–42). These epitopes are divided into three segments, numbered Epi1, Epi2 and Epi3 respectively, and their amino acid sequences are shown in SEQ ID NO: 2, 3, and 4; Epi1, Epi2, and Epi3 contain each epitope shown in Table 1, respectively.

[0063] Table 1 Epitope prediction results of Pasteurella septica toxin protein (PMT)

[0064] Divide into sections

sequence

SEQ ID NO.

...

Embodiment 2

[0069] Embodiment 2 The cultivation of Bordetella bronchiseptica and Pasteurella multocida

[0070] 1. Culture of Bordetella bronchiseptica (B.bronchiseptica)

[0071] In this example, Bordetella bronchiseptica (B. bronchiseptica) was distributed from the Animal Health Laboratory of the Council of Agriculture, Executive Yuan (Taiwan).

[0072]First inoculate the Bordetella bronchiseptica (B. bronchiseptica) on TSB solid medium [containing 5% (v / v) yeast extract, 10% (v / v) serum, soybean decomposition Protein casein medium (tryptic soy broth, TSB, BD Company, USA)], after culturing overnight at 37°C, a single colony was selected and inoculated in brain heart infusion (BHI) liquid medium (BD Company, USA) ), shake culture overnight at 37°C; then take the bacterial solution and inoculate it in BHI liquid medium, culture shake overnight at 37°C, and calculate the colony forming unit (CFU) value; finally add formaldehyde (formaldehyde ), shaking at room temperature for 24 to 36 h...

Embodiment 3

[0076] Example 3 Preparation of the porcine atrophic rhinitis immune composition containing Pasteurella septica toxin recombinant protein (rPMT)

[0077] The Pasteurella septica toxin recombinant protein (rPMT) (SEQ ID NO:10 or 11) (final concentration is 65 μ g / ml) obtained in embodiment one and the inactivated bronchial septic Bordet obtained in embodiment two B. bronchiseptica (final concentration 1x10 9 CFU / ml), inactivated Pasteurella septica type A (PmA) (final concentration 1x10 9 CFU / ml) and inactivated Pasteurella septica type D bacteria (PmD) (final concentration is 1x10 9 CFU / ml) was mixed evenly with phosphate buffered solution (PBS), and aluminum gel [final concentration was 30% (v / v)] was added as an adjuvant to prepare a porcine atrophic rhinitis immune composition.

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PUM

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Abstract

The present invention relates to a Pasteurella multocida toxin recombinant protein and use thereof, the recombinant protein comprising at least one epitope of Pasteurella multocida toxin recombinant protein, and when the number of the epitopes is a plurality, the epitopes can be connected by a connector therebetween. The recombinant protein can further comprise the amino acid sequence of at least one unit of a complement cleavage C3d, and the epitopes and the amino acid sequence of the complement cleavage C3d can be connected by a connector. The present invention also relates to a nucleotide sequence encoding the above-mentioned Pasteurella multocida toxin recombinant protein, and a porcine atrophic rhinitis immunogenic composition comprising the above-mentioned Pasteurella multocida toxin recombinant protein.

Description

technical field [0001] The invention relates to the application of the Pasteurella septica toxin recombinant protein in the field of animal health care, in particular to the application of the porcine atrophic rhinitis immune composition containing the Pasteurella septica toxin recombinant protein in the field of animal health care. Background technique [0002] Atrophic Rhinitis (AR) is one of the three major infectious diseases of swine respiratory system. Porcine atrophic rhinitis causes facial bone deformities and chronic suppurative rhinitis in infected pigs, resulting in atrophy of the turbinates. In severe infection, lesions will also occur in the nasal cavity, jaw, and maxilla. The downstream fossa of the inferior turbinate is most commonly infected. The superior turbinate, inferior turbinate, nasal septum, or ethmoid bone can all be infected. [0003] Porcine atrophic rhinitis (AR) is caused by Bordetella bronchiseptica and Pasteurella multocida type A (PmA) and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/285C07K16/12C12N15/31A61K39/102A61K39/116A61K39/295A61P31/04A61P11/02G01N33/68G01N33/569
CPCA61K39/116G01N33/53C07K14/285C07K19/00A61K39/102A61K39/00C07K14/472C07K16/1242C07K2319/00G01N33/56911G01N33/68G01N2333/285A61K2039/552A61K2039/6031A61P11/02A61P31/04
Inventor 杨滢臻郭村勇
Owner 维克香港贸易有限公司
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