Porcine contagious pleuropneumonia actinobacillus serum 1-type zinc ion binding protein A deletion strain, construction method therefor and applications thereof

A technology of zinc ion binding and pleuropneumonia, applied in the biological field, can solve problems such as difficulty in disease prevention and treatment, excessive drug residues in public health, and increased risk of multi-drug resistance

Inactive Publication Date: 2014-02-19
INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
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Problems solved by technology

Although inactivated vaccines can effectively control the infection of homologous strains, due to the large number of serotypes and complex antigenic components, the control effect on heterologous strains is not satisfactory
The abuse of antibiotics has led to the...

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  • Porcine contagious pleuropneumonia actinobacillus serum 1-type zinc ion binding protein A deletion strain, construction method therefor and applications thereof
  • Porcine contagious pleuropneumonia actinobacillus serum 1-type zinc ion binding protein A deletion strain, construction method therefor and applications thereof
  • Porcine contagious pleuropneumonia actinobacillus serum 1-type zinc ion binding protein A deletion strain, construction method therefor and applications thereof

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Embodiment

[0024] 1. Materials

[0025] 1. Tool enzymes and reagents

[0026] Various restriction endonucleases, Ex / LA Taq DNA polymerase, T4 DNA ligase, Klenow fragment and other tool enzymes, DNA Marker, and RNase enzymes were purchased from Dalian Bao Biological Company.

[0027] DNA recovery kit was purchased from Tiangen Company.

[0028] Tryptic Soy Agar (TSA) and Tryptic Soy Broth (TSB) were purchased from Becton Dickinson and Company, USA.

[0029] Diaminopimelic acid (DAP), nicotinamide adenine dinucleotide (Nicotinamide Adenine Dinucleotide, NAD+), HRP-labeled goat anti-mouse / pig IgG secondary antibody and sucrose for negative screening were purchased from Sigma, USA .

[0030] Newborn bovine serum was purchased from Hangzhou Sijiqing Material Co., Ltd. and inactivated at 56°C for 30 min before use.

[0031] Ampicillin (Amp), kanamycin (Kan), chloramphenicol (Chloramphenicol, Cm), lysozyme, and proteinase K were all purchased from Invitrogen.

[0032] 2. Culture medium, an...

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Abstract

The invention relates to a porcine contagious pleuropneumonia actinobacillus serum 1-type zinc ion binding protein A (znuA) deletion strain, a construction method therefor and applications thereof. The microbial colllection number is CCTCC No: M2012476. The classification and nomenclature is inobacillus pleuropneumoniae APP-1( delta znuA). The collection time is November 23th, 2012. The collection place is China Center for Type Culture Collection. The collection address is Wuhan University, Wuhan, China. The znuA gene deletion mutantion strain is compared to the parent strain to study biological characteristics and pathogenic immunogenicity. The discoveries show that the pathogenicity of the gene deletion strain is obviously weaker than that of the parent strain APP. The gene deletion strain can be used for developing gene deletion inactivated vaccines of APP, and provides effective technologies and methods for preventing and treating porcine contagious pleuropneumonia safely and effectively.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a deletion strain of Actinobacillus pleuropneumoniae serum type 1 zinc ion binding protein A (znuA) and its construction method and application. Background technique [0002] Porcine contagious pleuropneumonia (PCP) is a highly contagious respiratory disease caused by Actinobacillus pleuropneumoniae (APP). The disease mainly causes hemorrhagic and necrotizing pneumonia accompanied by pleurisy in pigs, and most of them are the most acute or acute course and quickly die. The disease can occur in pigs of any age and is prevalent all over the world. Once an outbreak occurs, the economic loss will be huge. The current domestic control of porcine pleuropneumonia mainly relies on inactivated vaccine immunization and antibiotic treatment. Although inactivated vaccines can effectively control the infection of homologous strains, due to the large number of serotypes and complex a...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/09A61K39/02A61P31/04C12R1/01
Inventor 袁芳艳田永祥刘泽文周丹娜杨克礼段正赢郭锐
Owner INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
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