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Recombinant bordetella bronchiseptica strain, vaccine and use

A technology of Bordetella blood and bronchi, which is applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve biological safety problems and other problems, and achieve the effect of convenient immune operation, broad market application prospects, and good immune protection

Active Publication Date: 2015-05-27
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of the research results reported so far use expression plasmids carrying resistance genes, which are not accepted by people due to biological safety issues

Method used

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  • Recombinant bordetella bronchiseptica strain, vaccine and use
  • Recombinant bordetella bronchiseptica strain, vaccine and use
  • Recombinant bordetella bronchiseptica strain, vaccine and use

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Embodiment 1

[0041] 1. Primer design and PCR amplification

[0042] According to the reported whole genome sequence of Bordetella bronchiseptica RB50 (Parkhill J., 2003, Nature) (refer to the gene sequence of Genbank accession number AF315119), two pairs of primers were designed (all primers were provided by Nanjing GenScript Biotechnology Co., Ltd. Synthesis) to amplify the upstream homology arm and downstream homology arm of the aroA gene respectively, the size of the amplified fragments are 1388bp and 1383bp respectively, design Kpn I and Sal I restriction sites at both ends of the upstream arm, and design Hind at both ends of the downstream arm III and Sac I restriction sites. According to the reported toxin-producing Pasteurella multocida strain HN-13 (Tang Xianchun. Isolation, Identification and Biological Characteristics of Pasteurella multocida. [Master's Dissertation], CNKI http: / / www .cnki.net / kcms / detail / detail.aspx?dbcode=cmfd&dbname=cmfd2005&filename=2004138308.nh&uid=WEEvREc...

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Abstract

The invention belongs to the technical field of animal bacteriology and vaccine genetic engineering preparation, and relates to a resistance mark-free recombinant bordetella bronchiseptica strain for expression of a pig toxaigenic pasteurella multocida toxA gene fragment, and its vaccine containing the resistance mark-free recombinant bordetella bronchiseptica strain, a preparation method and a use. The resistance mark-free recombinant bordetella bronchiseptica strain QH0814 delta aroA / toxA-N for expression of the pig toxaigenic pasteurella multocida toxA gene fragment does not contain a 5-enolpyrul-shikimate-3-phosphate synthase aroA gene of bordetella bronchiseptica, and contains the pig toxaigenic pasteurella multocida toxA gene fragment. The invention also discloses the preparation method of the resistance mark-free recombinant bordetella bronchiseptica strain and the vaccine use. The genetic engineering vaccine can stimulate a pig to produce a protective immune response for resisting pig toxaigenic pasteurella multocida and a wild strain of pig bordetella bronchiseptica, and can effectively prevent the double infection caused by pig toxaigenic pasteurella multocida and pig bordetella bronchiseptica.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering of animal bacteria, and in particular relates to the construction of a recombinant Bordetella bronchiseptica live vaccine strain expressing the toxA gene fragment of Pasteurella multocida without a resistance marker, and a vaccine containing the strain Vaccine, its preparation method and application. Background technique [0002] Bordetella Bronchiseptica (Bb) is a Gram-negative bacterium with a wide host range that can infect most mammals and birds. This bacterium is one of the main pathogenic bacteria that cause atrophic rhinitis (AR) in pigs. Bb alone often causes recessive infection in pigs. The clinical manifestations of diseased pigs are slow growth and fattening delay, which greatly reduces feed remuneration and Production efficiency; and pig atrophic rhinitis caused by the joint action of Bb and toxin-producing Pasteurella multocida (Pm) is clinically characterized by rhinitis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21A61K39/116A61K39/102A61K39/10A61P31/04C12R1/01
Inventor 吴斌罗芬陈焕春吴磊曾松林侯康炜汤细彪余腾张佳思
Owner HUAZHONG AGRI UNIV
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