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Fusion protein containing mycoplasma hyopneumoniae antigen, vaccine composition and application

A technology of mycoplasma hyopneumoniae and vaccine composition, which is applied to medical preparations containing active ingredients, vaccines, veterinary vaccines, etc., can solve the problems of difficult cultivation of mycoplasma hyopneumoniae, imperfect separation technology, long growth cycle, etc., and achieve Convenient for large-scale production, excellent protective effect, and good immune protection effect

Active Publication Date: 2018-04-03
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although vaccines are widely used, there are still some problems with their effects. Recent studies have shown that the development of inactivated vaccines is based entirely on adjuvanted whole-cell preparations or membrane preparations, and the immune effect is not completely effective. Although vaccine immunization can reduce lung damage, However, it cannot prevent the proliferation and spread of Mycoplasma hyopneumoniae in the lungs
On the other hand, the cultivation and isolation technology of Mycoplasma hyopneumoniae is not perfect at present in China. The cultivation of Mycoplasma hyopneumoniae is difficult and the growth cycle is long, which leads to high production cost of traditional vaccines.
[0004] The existing inactivated Mycoplasma hyopneumoniae vaccines often require two immunizations to achieve good results, and the process of strengthening immunization brings economic and procedural burdens to users

Method used

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  • Fusion protein containing mycoplasma hyopneumoniae antigen, vaccine composition and application
  • Fusion protein containing mycoplasma hyopneumoniae antigen, vaccine composition and application
  • Fusion protein containing mycoplasma hyopneumoniae antigen, vaccine composition and application

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Embodiment approach

[0037] As an embodiment of the present invention, the content of the fusion protein in the vaccine composition is ≥100 μg / ml.

[0038] As a preferred embodiment of the present invention, the content of the fusion protein in the vaccine composition is 100-400 μg / ml.

[0039]As a more preferred embodiment of the present invention, the content of the fusion protein in the vaccine composition is 250 μg / ml.

[0040] The term "mycoplasma pneumoniae vaccine composition" used in the present invention refers to a vaccine capable of preventing and / or treating diseases or diseases related to infection by Mycoplasma hyopneumoniae. The vaccine that can be used for the vaccine composition of mycoplasma pneumoniae in swine includes (but not limited to) complete or partial mycoplasma pneumoniae cell preparations, inactivated or improved live vaccines, and subunit vaccines.

[0041] The Mycoplasma hyopneumoniae vaccine composition of the present invention is preferably a subunit vaccine, the ...

Embodiment 1

[0082] Example 1, Expression, Identification and Purification of Fusion Protein LTB-P97R1

[0083] 1.1 Construction and identification of LTB-P97R1 fusion gene cloning vector

[0084] According to NCBI ( http: / / www.ncbi.nlm.nih.gov ) gene sequence (see SEQ ID No.1) of enterotoxigenic E. The P97R1 protein gene sequence (see SEQ ID No.3), the primer sequences of LTB protein and P97R1 protein were designed respectively with Primer Premier 5 software, and the upstream primer of LTB protein and the downstream primer of P97R1 protein were respectively added with EcoRI and BamHI digestion site, and the PCR method was used to amplify Escherichia coli heat-labile enterotoxin B subunit (LTB) and Mycoplasma hyopneumoniae P97R1 protein. Details are as follows:

[0085] LTB gene primers were designed as follows:

[0086] Upstream primer: 5'-CGCGAATTCGTCCCCAGACTATTACA-3'

[0087] Downstream primer: 5'-TATACCCATACTGATTG CCGCAAT TGA-3'

[0088] P97R1 gene primers were designed as follow...

Embodiment 2

[0101] Embodiment 2, the preparation of fusion protein LTB-NrdFR2

[0102] According to NCBI ( http: / / www.ncbi.nlm.nih.gov ) gene sequence (see SEQ ID No.1) of enterotoxigenic E. The NrdF protein gene sequence (see SEQ ID No.5), use Primer Premier 5 software to design the primer sequences of LTB protein and NrdF protein respectively, and add EcoRI, HindIII enzyme digestion respectively at the upstream primer of LTB protein and the downstream primer of NrdFR2 protein site, and the PCR method was used to amplify Escherichia coli heat-labile enterotoxin B subunit (LTB) and Mycoplasma hyopneumoniae NrdFR2 protein. Details are as follows:

[0103]LTB gene primers were designed as follows:

[0104] Upstream primer: 5'-CGCGAATTCGTCCCCAGACTATTACA-3'

[0105] Downstream primer: 5'-TATACCCATACTGATTG CCGCAAT TGA-3'

[0106] NrdFR2 gene primers were designed as follows:

[0107] Upstream primer: 5'-GATCTATTATATAAACTAATTG-3'

[0108] Downstream primer: 5'-CCCAAGCTTTTAAAACTCCCAATTCTT...

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Abstract

The invention relates to a fusion protein. The fusion protein comprises enterotoxigenic escherichia coil heat-labile enterotoxin B subunit and mycoplasma hyopneumoniae antigen protein in a sequence from an N end to a C end. The invention also relates to a vaccine composition prepared from the fusion protein and an application thereof. The vaccine composition provided by the invention can achieve the immune effect of the commercial inactivated vaccine or better immune effect after one-time immunization.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to fusion proteins containing mycoplasma hyopneumoniae antigens, vaccine combinations and applications. Background technique [0002] Mycoplasma pneumonia of swine (MPS) is a chronic, contagious disease caused by Mycoplasma hyopneumoniae (Mhp). Porcine respiratory disease complex (PRDC) is one of the important pathogens. The disease has endangered the pig industry for a long time and is widely distributed in countries all over the world. Clinically, it is characterized by cough, wheezing and typical "shrimp meat"-like lesions in the lungs, with high morbidity and low mortality. [0003] At present, the control of Mycoplasma pneumonia mainly depends on the application of vaccine immunization and antibiotics. Although the use of antibiotics has a certain effect in the treatment of asthma, the country has paid more and more attention to the abuse o...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K39/02A61P31/04
CPCA61K39/0241A61K2039/552A61K2039/55566A61K2039/572A61K2039/575C07K14/245C07K14/30C07K2319/55
Inventor 田克恭朱巧艳孙进忠张许科
Owner PU LIKE BIO ENG
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