Bordetella bronchiseptica gene deleted vaccine and application

A gene deletion vaccine, Bordetella blood technology, applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problems of no product coming out, achieve the effect of reducing virulence, protecting against attacks, and broad market application prospects

Inactive Publication Date: 2009-11-11
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, foreign scholars have begun to study Bordetella bronchiseptica as a live attenuated vaccine, but so far there is no relatively mature product.

Method used

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  • Bordetella bronchiseptica gene deleted vaccine and application
  • Bordetella bronchiseptica gene deleted vaccine and application
  • Bordetella bronchiseptica gene deleted vaccine and application

Examples

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Embodiment 1

[0033] 1. aroA primer design and PCR amplification

[0034] According to the reported whole genome sequence of Bordetella bronchiseptica RB50 (refer to the gene sequence of GenBank accession number AF315119), two pairs of primers were designed (all primers were synthesized by Shanghai Sangon Bioengineering Technology Co., Ltd.) to amplify the upstream of aroA gene respectively. Homology arm (SEQ NO.3) and downstream homology arm (SEQ NO.1), the amplified fragment sizes are 863bp and 900bp respectively, KpnI and BamHI restriction sites are designed at both ends of the upstream arm, and respectively designed at both ends of the downstream arm BamHI and SacI restriction sites. The primer sequences are as follows:

[0035] C1:5`-TGC GGTACC TAGGTTGGAATCCATTTGGC-3`(Kpn I)

[0036] C2: 5`-TAA GGATCC CCATGGCTATTTCCGCATCC-3`(BamH I)

[0037] Primers C1 and C2 amplify the upstream homology arm 863bp

[0038] T1: 5`-TGT GGATCC TGGAATCCTTGCGCCAATTG-3`(BamH I)

[0039] T2: 5`-TA...

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Abstract

The invention belongs to the technical field of vaccine preparation of animal gene engineering and particularly relates to the construction of a Bordetella bronchiseptica aroA (5-enolpyruvyl-shikimate-3-phosphate synthase) gene deleted bacterial strain, and the preparation and application of a gene deleted vaccine. The Bordetella bronchiseptica gene deleted bacterial strain QH0814DeltaaroA is preserved in the China Center for Type Culture Collection with the collection number of CCTCCNO: M208018. The deletion of 5-enolpyruvyl-shikimate-3-phosphate synthase (aroA) gene with the full length of 1340bp causes an obstacle when the bacterial strain metabolizes aromatic amino acid. The invention prepares the Bordetella bronchiseptica gene deleted vaccine by using the gene deleted bacterial strain. The invention also discloses the application of the gene deleted bacterial strain in preparing the Bordetella bronchiseptica gene deleted vaccine (attenuated live vaccine).

Description

technical field [0001] The invention belongs to the technical field of genetic engineering of animal bacteria, and in particular relates to a Bordetella bronchiseptica 5-enolpyruvylshikimate-3-phosphate synthase gene (aroA) deletion strain, a gene deletion vaccine and application thereof. Background technique [0002] Atrophic rhinitis (Atropic Rhinitis, AR) is a porcine respiratory infectious disease caused by the joint action of Bordetella Bronchiseptica (Bb) and toxin-producing Pasteurella multocida (Pm). The disease has caused serious economic losses to the pig industry in the world, and the World Organization for Animal Health (OIE) and my country have listed it as a B or B animal disease. Porcine atrophic rhinitis is clinically characterized by rhinitis, turbinate atrophy, and decreased growth performance. Studies have shown that Bordetella bronchiseptica is the primary pathogen that causes atrophic rhinitis in pigs. Bb alone often causes recessive infection. The disea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/10A61P31/04C12R1/01
Inventor 吴斌胡睿铭何华李伦勇汤细彪卢顺陈焕春金梅林何启盖
Owner HUAZHONG AGRI UNIV
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