Avian pathogenic escherichia coli strain and application thereof in vaccine preparation
A technology of avian pathogenic Escherichia coli, which is applied in the field of avian pathogenic Escherichia coli strains and its application in vaccine preparation, can solve problems such as drug residues in poultry products and affect the safety of poultry products, and achieve good public health significance, The effect of reduced use
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0023] Example 1 Construction of avian pathogenic Escherichia coli luxS and aroA deletion strains
[0024] Using the genome of avian pathogenic Escherichia coli DE17 as a template and luxS-UF / luxS-UR as primers, the upstream homology arm fragment of the luxS gene (775bp) was amplified, and luxS-DF / luxS-DR was used as primers to amplify Amplification of the homology arm fragment downstream of the luxS gene (1023bp). Use aroA-UF / aroA-UR as primers to amplify the upstream homology arm fragment of aroA gene (886bp), and use aroA-DF / aroA-DR as primers to amplify the downstream homology arm fragment of aroA gene (957bp ).
[0025] The upper and lower homology arm fragments of the luxS and aroA genes were cloned into the pUC19 vector to construct pUC19-luxS-up-down and pUC19-aroA-up-down recombinant vectors, respectively.
[0026] The pKD3 plasmid was used as a template and pkD3-F / pkD3-R were used as primers to amplify the chloramphenicol resistance gene (cat). After the PCR produc...
Embodiment 2
[0035] Embodiment 2 median lethal dose (LD 50 ) determination
[0036] In this example, the wild strain DE17, the deletion strains DE17ΔaroA and DE17ΔaroAΔluxS were cultured to the logarithmic phase, the bacteria were collected, washed three times with sterile PBS, the number of bacteria was adjusted, and the ratio was diluted. Each strain of bacteria is divided into five groups, with 8 cherry valley ducks in each group, so that the number of bacteria in each strain of bacteria and each group is 10 7 CFU / piece, 10 6 CFU / piece, 10 5 CFU / piece, 10 4 CFU / piece, 10 3 CFU / only and 10 2 CFU / only was injected into the thigh muscle, observed for 7 days after the challenge, and recorded the death of Cherry Valley duck. Data statistics using Reed-Muench method to calculate LD 50 .
[0037] The detection results showed that the LD of DE17, the deletion strain DE17ΔaroA and the double deletion strain DE17ΔluxSΔaroA 50 1.2×10 4 CFU, 3.8×10 5 CFU and 2.4×10 6 CFU. After the del...
Embodiment 3
[0038] Example 3 Effect of aroA and luxS deletion on APEC motility
[0039] In this example, the wild strain DE17, the deletion strain DE17ΔaroA and the double deletion strain DE17ΔluxSΔaroA were cultivated to the logarithmic phase, the bacteria were washed three times with sterile PBS, and the OD 600 Adjust to 1.0. In the middle of the motility medium, add 10 μl of bacteria, and culture overnight in a 37°C incubator. Use a vernier caliper to record the size of the bacterial circle for comparative analysis. The results of motility analysis showed that the deletion of aroA gene alone did not affect the motility of APEC, while the simultaneous deletion of luxS and aroA genes could significantly reduce the motility of APEC (pfigure 1 shown.
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com