Vaccine for differentiating porcine actinobacillus pleuropneumonia serum 7-type double-gene deletion mutant of vaccine immunity and virus infection animal and application thereof

A technology of porcine pleuropneumoniae and Actinobacillus, which is applied in the fields of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problems of no relevant reports in in-depth research, late discovery and research start of toxin ApxIV, etc. force reduction effect

Inactive Publication Date: 2008-09-17
HUAZHONG AGRI UNIV
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the three toxins ApxI, ApxII and ApxIII, the discovery and research of the toxin ApxIV started relatively late. Most of the current research on ApxIV is based

Method used

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  • Vaccine for differentiating porcine actinobacillus pleuropneumonia serum 7-type double-gene deletion mutant of vaccine immunity and virus infection animal and application thereof
  • Vaccine for differentiating porcine actinobacillus pleuropneumonia serum 7-type double-gene deletion mutant of vaccine immunity and virus infection animal and application thereof
  • Vaccine for differentiating porcine actinobacillus pleuropneumonia serum 7-type double-gene deletion mutant of vaccine immunity and virus infection animal and application thereof

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Embodiment 1

[0045] 1. Toxin apxIIC primer design and PCR amplification

[0046] According to the reported APP-7 strain sequence (with reference to the gene sequence of GenBank accession number AF315119), two pairs of primers were designed to respectively amplify the upstream arm and downstream arm of the activation gene apxIIC of the toxin apxII, and the sizes of the amplified fragments were 1600bp and 1800bp respectively. PstI and EcoRI restriction sites were designed at both ends of the upstream arm, and KpnI and XhoI restriction sites were designed at both ends of the downstream arm. The above primers were synthesized by Shanghai Bioengineering Company. The primer sequences are as follows:

[0047] pII-1: 5'GCCTGCAGATTAAACAGCACCCTAC-3' (PstI)

[0048] pII-2: 5'-GTGAATTCGTAGCATCATCCCTCCC-3'(EcoRI) upper arm 1600bp

[0049] pII-3: 5'-TCCTCGAGGGCAATTAGAATCTATC-3' (EcoRI)

[0050] pII-4: 5'-GCGGTACCTTCACCTGGAGTTAGT-3'(KpnI) lower arm 1800bp

[0051] 2. Cloning of the upstream arm and ...

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Abstract

The invention belongs to the animal bacteria genetic engineering field, and relates particularly to construction of a vaccine for a serotype-7 double gene deletion mutation strain for differentiating actinobacillus pleuropneumoniae of swine of a vaccine immunized animal and a wild virus infected animal, and the application of the strain. In the invention, a serotype-7 double gene deletion mutation strain APP-7-mut01 for differentiating actinobacillus pleuropneumoniae of swine of a vaccine immunized animal and a wild virus infected animal (with accession number of CCTCC NO:M207004) is obtained. The strain deletes two main virulence factors of apxIIC and apxIVAN of APP serotype-7. The mutant strain is safe for swines, and has toxins of ApxIIA and ApxIVAC protein, thereby ensuring the immunity efficacy of the vaccine. The vaccine of the mutant strain is used to immunize animals, so that antibodies aiming at Apx IVAN protein are no longer produced inside the animals, however wild virus infection can produce antibodies aiming at ApxIVAN. The invention further discloses the construction, the vaccine preparation and the application of the double gene deletion mutation strain.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering of animal bacteria, and in particular relates to the construction, vaccine preparation and application of an actinobacillus pleuropneumoniae serum type 7 gene deletion mutant strain for distinguishing between vaccine-immunized and wild virus-infected animals. Background technique [0002] Porcine contagious pleuropneumonia (PCP) is a porcine contagious respiratory disease caused by Actinobacillus pleuropneumoniae (APP), which has caused serious economic losses to the world's pig industry. Since the prevalence of PCP was discovered in our country, the vast number of veterinary workers have conducted a series of researches on its etiology, epidemiology, diagnosis and control, and achieved certain achievements. But at present, the incidence of this disease in our country is still increasing year by year, and the positive rate of some pig farms has reached more than 70%, which has become o...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/02A61P31/04
Inventor 贝为成陈焕春刘金林周锐何启盖金梅林方六荣吴斌肖少波曹胜波
Owner HUAZHONG AGRI UNIV
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