Method for preparing actinobacillus pleuropneumoniae (App) bacterial ghost and method for preparing subunit vaccine by loading pasteurella antigen with App bacterial ghost

A subunit vaccine, Pasteurella technology, applied in bacterial antigen components, botanical equipment and methods, biochemical equipment and methods, etc., can solve problems such as poor cross-protection ability, incomplete cleavage, and lack of antigenic epitopes

Inactive Publication Date: 2011-01-05
TIANJIN AGRICULTURE COLLEGE
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  • Summary
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  • Description
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Problems solved by technology

All types of vaccines have advantages and disadvantages: inactivated vaccines are low in cost, safe, and convenient, but have poor cross-protection ability; attenuated vaccines have good cross-protection and immune effects, but there is a risk of reversion; subunit vaccines are safe and easy to prepare, but they need to be compared with Appropriate adjuvant can exert the best effect, and the cross-protectio

Method used

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Embodiment Construction

[0043] Below in conjunction with specific embodiment the present invention is described in further detail:

[0044] 1 Materials and methods

[0045] 1.1 Materials

[0046] Restriction endonucleases EcoRI, SalI, ApaI and BamHI, PhiX174 phage RF I plasmid, pMC-Express and pBV220 plasmid were purchased from Fermentas MBI Company, USA. The cloning vector pMD18-T and E.coli DH5α were purchased from Dalian TaKaRa Company. Staphylococcus aureus standard strain ATCC25923 and App 1 Shope 4074 strain were purchased from China Veterinary Drug Control Institute.

[0047] 1.2 Method

[0048] 1.2.1 Design and synthesis of primers

[0049] Two pairs of primers were designed according to the coding sequences of phage PhiX174 cleavage gene E (sequence number: 9626372) and Staphylococcus aureus nuclease A gene SN (sequence number: 21281729) in GenBank:

[0050] E1: 5'-CAG GAATTC ATGGTACGCTGGACTTTGTGG-3' (EcoRI)

[0051] E2: 5'-GGC GTC GAC TTACTCCTTCCGCACGTAAT-3'(SalI)

[0052] SN1: ...

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Abstract

The invention discloses a method for preparing an actinobacillus pleuropneumoniae (App) bacterial ghost and a method for preparing a subunit vaccine by loading a pasteurella antigen with the App bacterial ghost. A recombinant swine App bacterial ghost is prepared by controllable double-cracking technology and a pasteurella protection gene is introduced into an App bacterial ghost carrier, so that swine pleuropneumonia and a pasteurella bigeminal gene vaccine for preventing and treating swine pasteurellosis and swine pleuropneumonia are obtained. The preparation of the bacterial ghost carrier and the application of the bacterial ghost carrier to the prevention and treatment of important animal epidemic diseases are realized and a method is provided for the research of a multi-geminal gene vaccine at the same time. An animal experiment indicates that the protection rates of the bigeminal vaccine on infectious swine pleuropneumonia and pasteurellosis are up to 99 percent and 99.2 percent respectively.

Description

technical field [0001] The invention relates to the field of biotechnology, specifically, a method for preparing App bacteria ghosts by using E.coli-App shuttle carrier pMC-Express and double gene cleavage, and using App bacteria ghosts to load Pasteurella antigen OmpH outer membrane protein to prepare sub The unit vaccine approach. Background technique [0002] Actinobacillus pleuropneumoniae (App) and Pasteurella multocida are common and important pathogenic bacteria in the respiratory tract of pigs. The clinical symptoms caused by them are difficult to distinguish, and they are often mixed infections. A large number of epidemiological survey data show that due to the extensive use of antibacterial drugs, drug resistance of Actinobacillus pleuropneumoniae and Pasteurella bacteria is widespread, especially the emergence of highly resistant strains, resulting in the clinical prevalence of most drugs on drug-resistant bacteria. Decreased or ineffective efficacy. Currently c...

Claims

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Application Information

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IPC IPC(8): A61K39/116A61K39/102A61K39/02A61K39/39A61P31/04C12N15/33C12N15/31C12N15/55C12N15/74C12N1/21
Inventor 金天明孙颖刘田生高静
Owner TIANJIN AGRICULTURE COLLEGE
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