Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

APEC double-gene rpoS and arcA deletion strain and attenuated vaccine

A kind of attenuated vaccine and double-gene technology, applied in the field of microorganisms, can solve the problems of poor immune effect and inconvenient immune route

Inactive Publication Date: 2021-04-23
NANTONG UNIVERSITY
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although traditional inactivated vaccines and subunit vaccines have been used, they are gradually replaced by attenuated live vaccines due to poor immune effects and inconvenient immunization routes.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • APEC double-gene rpoS and arcA deletion strain and attenuated vaccine
  • APEC double-gene rpoS and arcA deletion strain and attenuated vaccine
  • APEC double-gene rpoS and arcA deletion strain and attenuated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Construction of APEC double gene rpoS and arcA deletion strain

[0018] The rpoS and arcA double gene deletion strain of avian pathogenic Escherichia coli in the present invention is derived from a wild-type virulent strain of avian pathogenic Escherichia coli, the strain name is FY26, which is isolated from chickens suffering from colibacillosis. (Zhuge X, Sun Y, JiangM, Wang J, Tang F, Xue F, Ren J, Zhu W, Dai J. Acetate metabolic requirement of avian pathogenic Escherichia coli promotes its intracellular proliferation within macrophage. Veterinary research. 2019 Dec 1; 50(1) : 31.) The present invention mainly adopts the method of Red homologous recombination to knock out rpoS and arcA double gene in described avian pathogenic Escherichia coli one by one, construct a kind of rpoS and arcA double gene deletion avian pathogenic Escherichia coli .

[0019] The sequence of the deleted rpoS gene is shown in SEQ ID NO.1.

[0020] The sequence of the deleted ar...

Embodiment 2

[0032] Example 2: The effect of deletion of rpoS and arcA genes on the pathogenicity of avian pathogenic Escherichia coli

[0033] In order to verify the attenuation effect of the deletion strain of the present invention on avian pathogenic Escherichia coli infection, through a variety of animal infection tests, including chicken embryo infection test, chick infection test and duckling infection test, etc., the wild strain and the deletion strain were determined to chicken. The effect of lethal dose. The effect of the wild strain and the deletion strain on the ability to infect DF-1 cells was tested, as well as the level of bacterial colonization in various viscera when infected chicks were tested. The results showed that the deletion of rpoS and arcA genes significantly reduced the pathogenicity of avian pathogenic Escherichia coli, and the deletion strain FY26ΔrpoS / arcA was an attenuated strain.

[0034] 1 Chick embryo lethal test

[0035] The chicken embryo lethality test...

Embodiment 3

[0063] Embodiment 3: Avian pathogenic Escherichia coli rpoS and arcA double gene deletion vaccine

[0064]The obtained avian pathogenic Escherichia coli rfaH gene deletion strain FY26ΔrpoS / arcA was identified, and each generation was inoculated on LB agar medium to observe the colony color, and the gene of avian pathogenic Escherichia coli was used for detection to identify the missing bacteria. genetic stability. After sub-passaging, it was found that FY26ΔrpoS / arcA maintained the phenotypic characteristics of gene deletion, and the identified gene was still missing, and heredity was stable. The rpoS and arcA double gene deletion strain FY26ΔrpoS / arcA was cultured on solid medium, and a single colony was picked and cultured in liquid medium until the concentration of viable bacteria reached 5.0×10 9 CFU / mL. The preparation method of the gelatin protectant is as follows: add 40g of sucrose and 9g of gelatin to every 100mL of deionized water, after fully melting, sterilize at...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of microorganisms, and discloses an APEC double-gene rpoS and arcA deletion strain and an attenuated vaccine. A construction method of the APEC double-gene rpoS and arcA deletion strain is to knock out rpoS and arcA double genes by a Red homologous recombination method to obtain avian pathogenic Escherichia coli FY26 delta rpoS / arcA which is preserved in China Center for Type Culture Collection with the preservation number of CCTCC M 2020613. The deletion strain has good biological safety and immunogenicity, is low in toxicity, and can be used for preparing avian pathogenic escherichia coli attenuated vaccines to prevent avian pathogenic escherichia coli infection. The attenuated vaccine prepared from the avian pathogenic escherichia coli FY26 delta rpoS / arcA disclosed by the invention has immune protection on avian pathogenic escherichia coli infection and is good in toxin attacking immune protection effect.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to an APEC double-gene rpoS and arcA deletion strain and an attenuated vaccine. Background technique [0002] Avian pathogenic Escherichia coli (Avian pathogenic E.coli, APEC) is an important bacterial pathogen that endangers the poultry industry, and it belongs to extraenteric pathogenic Escherichia coli (ExPEC). Avian pathogenic Escherichia coli mainly infects poultry through the respiratory tract, leading to multi-system mixed infection of poultry, and even acute death of poultry, causing serious losses to the poultry industry and posing a threat to food safety. The pathogenic subtypes of extraintestinal Escherichia coli are named as urethral pathogenic Escherichia coli (UPEC), sepsis Escherichia coli (SEPEC), neonatal meninges coli (NMEC) and avian pathogenic Escherichia coli. The evolution relationship between avian pathogenic Escherichia coli and clinical isolates o...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N1/21C12N15/70A61K39/108A61P31/04C12R1/19
CPCC07K14/245C12N15/70A61K39/0258A61P31/04A61K2039/522A61K2039/552
Inventor 诸葛祥凯戴建君姜敏周洲王忠星
Owner NANTONG UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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