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Salmonellapullorum attenuated mutant strain deltaS6702(SpiA) and construction method thereof

A technology of S6702 and Salmonella, which is applied in the attenuated Salmonella pullorum mutant strain △S6702 (SpiA) and its construction field, can solve the problems of inconvenient immunization route, endangering economic loss, poor immunization effect and the like

Inactive Publication Date: 2011-09-21
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease can be transmitted both vertically and seriously horizontally, so the damage and economic loss are huge
[0003] At present, the use of vaccines to prevent infectious diseases in modern medicine is the most effective measure. The research on the prevention of pullorum by vaccination has never been interrupted. replaced by live vaccines

Method used

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  • Salmonellapullorum attenuated mutant strain deltaS6702(SpiA) and construction method thereof
  • Salmonellapullorum attenuated mutant strain deltaS6702(SpiA) and construction method thereof
  • Salmonellapullorum attenuated mutant strain deltaS6702(SpiA) and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Construction of S6702 Gene Mutant

[0022] 1. Identification of S6702:

[0023] 1.1 Bacterial isolation

[0024] Use an inoculation loop to aseptically take a sample from the liver of a dead chicken suspected of pullorum disease, and inoculate it on a MacConkey agar plate, a gallbladder milk (DHL) agar plate, and a S.S. colony morphology. After culturing at 37°C for 24 hours, a colorless, translucent, neat-edged, smooth type was formed on MacConkey medium, gallic acid milk (DHL) agar medium and S.S medium (Qingdao High-tech Park Haibo Biotechnology Co., Ltd.) Small circular colonies, about 1.5mm in diameter. Pick suspicious colonies for further pure culture.

[0025] 1.2. Bacteria identification

[0026] 1.2.1 Microscopic examination of staining After Gram staining, observe the shape of the bacteria and measure the size of the bacteria under an ordinary optical microscope. The results of Gram staining showed that microscopic examination could observe sc...

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Abstract

The invention relates to a strain with the deletion of wild type Salmonellapullorum virulence genes, and a construction method of the strain. The Salmonellapullorum attenuated mutant strain deltaS6702(SpiA) is Salmonellapullorum CGMCC NO:4377. The strain is obtained by knocking out Salmonellapullorum SpiA genes by using a Red homologous recombination method. The Salmonellapullorum CGMCC NO:4377 can be used as a Salmonellapullorum attenuated candidate live vaccine in application.

Description

technical field [0001] The invention relates to a construction method for deleting the virulence gene of wild-type Salmonella pullorum, in particular to SpiA Gene as the target gene, and a primer design for gene deletion, electroporation method and method for eliminating resistance gene by expressing FLP recombinase auxiliary group plasmid, to obtain Salmonella pullorum attenuated mutant strain △S6702(SpiA) , deposit number CGMCC NO:4377. Background technique [0002] Pullorum pullorum is a septic infectious disease of chickens and turkeys caused by Salmonella pullorum pullorum. Salmonella pullorum mostly infects young chicks within 20 days of age, causing white diarrhea, and the mortality rate is extremely high. Adult chickens can carry the bacteria without clinical symptoms. The disease can be transmitted both vertically and seriously horizontally, so the harm and economic loss caused are huge. [0003] At present, the use of vaccines to prevent infectious diseases in m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/74A61K39/112A61P31/04A61P1/12C12R1/42
Inventor 彭大新陈素娟刘秀梵张小荣卢艳董洪燕
Owner YANGZHOU UNIV
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