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

Edwardsiella tarda immunogenic protective antigen, and related expression vector, vaccine and application

A technology of protective antigen and expression vector, applied in the field of immune protective antigen and antigen, can solve the problems of increasing the difficulty of antibiotic treatment and application limitations, and achieve the effect of being suitable for large-scale promotion and application.

Active Publication Date: 2011-10-05
EAST CHINA UNIV OF SCI & TECH
View PDF1 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As antibiotics are widely recognized, their use in aquaculture is increasingly limited
In addition, pathogenic Edwardsiella tarda is often found to be naturally resistant to multiple antimicrobial complexes, increasing the difficulty of antibiotic-based therapy

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
  • Edwardsiella tarda immunogenic protective antigen, and related expression vector, vaccine and application
  • Edwardsiella tarda immunogenic protective antigen, and related expression vector, vaccine and application
  • Edwardsiella tarda immunogenic protective antigen, and related expression vector, vaccine and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The preparation of embodiment 1 recombinant protein vaccine

[0026] 1. Experimental materials

[0027] 1. Strains and plasmids

[0028] Escherichia coli TOP10 and E. coli BL21 (DE3) were purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.; Edwardsiella tarda E. tarda EIB202, the preservation number is: CCTCC M 208068; pET 28a (+) vector Purchased from Bao Biological Engineering (Dalian) Co., Ltd.

[0029] 2. Medium and culture conditions

[0030] Medium:

[0031]LB: 1% Tryptone, 0.5% Yeast extract, 1% NaCl. Add 2% ager to the solid medium. Autoclave at 121°C for 20 minutes. For the cultivation of Escherichia coli.

[0032] TSB: 3% TSB. Add 2% ager to the solid medium. Autoclave at 121°C for 20 minutes. For EIB202 culture.

[0033] DHL medium: 60g of gallic acid milk agar medium was dissolved in 1L of deionized water, boiled repeatedly to dissolve until clear, and made a plate after cooling.

[0034] Training conditions:

[0035] Escherichia c...

Embodiment 2

[0056] Embodiment 2: Taking zebrafish as the experimental animal's immune protection test by injection

[0057] Protein Concentration Determination

[0058] The OD280 of the nucleic acid quantification instrument NanoDrop ND-1000 spectrophotometer was detected to determine the concentration of the recombinant protein, and the protein was diluted to a predetermined concentration with PBS buffer.

[0059] Immunization of zebrafish

[0060] The purified FD recombinant protein was mixed with the adjuvant ISA 763A at a ratio of 7:3, and the final protein concentration was 0.3 μg / μl. Zebrafish were cultured in random groups, 30 fish / group, and each zebrafish was immunized by tail intramuscular injection. Then feed them normally, observe the activity and death of the fish. Immunization time is one month. The negative control was the mixed group of PBS and adjuvant.

[0061] Before the injection operation, the zebrafish was soaked in 100ng / ml MS-222 for anesthesia. After the end...

Embodiment 3

[0070] Embodiment 3: Taking turbot as the experimental animal's immune protection test by injection

[0071] Immunization of turbot

[0072] Turbot were cultured in random groups, 30 fish / group, and each fish was intramuscularly injected into the tail at a dose of 100 μl / fish. After the purified protein was dialyzed overnight, according to its concentration, it was mixed with the adjuvant ISA 763A at a ratio of 7:3 until the protein concentration reached 0.3 μg / μl. PBS and adjuvant ISA 763A immunization group were negative controls. Then feed them normally, observe the fish's activities and whether they die. The immunization time is 10 weeks.

[0073] Before the injection operation, soak the turbot in 100ng / ml MS-222 for anesthesia. After the experimental period, the remaining turbot were euthanized by soaking in 300 ng / ml MS-222 for more than 10 minutes.

[0074] Turbot attack

[0075] The challenge method of turbot is the same as that of zebrafish. After the turbot is ...

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 provides an Edwardsiella tarda immunogenic protective antigen which is an Edwardsiella tarda flagellar related protein with an amino acid sequence expressed in SEQ ID NO:2 and a preferred nucleotide sequence expressed in SEQ ID NO:1, a recombination expression vector corresponding to the Edwardsiella tarda immunogenic protective antigen, an Edwardsiella tarda subunit vaccine prepared by the Edwardsiella tarda immunogenic protective antigen, and an application of the Edwardsiella tarda immunogenic protective antigen in the preparation of the Edwardsiella tarda subunit vaccine. The Edwardsiella tarda immunogenic protective antigen provided by the invention has a good immunogenicity and can be used for preparing a safe, effective and economic vaccine, thus has a high commercialvalue and is suitable for large scale popularization and application.

Description

technical field [0001] The present invention relates to the technical field of antigens, more specifically, to the technical field of immune protective antigens, in particular to an Edwardsiella tarda immune protective antigen, related expression vectors, vaccines and applications. Background technique [0002] With the continuous and steady development of fish farming, various disease problems have become increasingly prominent, which have seriously affected the production and growth of aquaculture. In my country, sudden and explosive diseases of seawater cage culture and factory culture fish developed in recent years occur frequently. At present, the average mortality loss rate of mariculture in my country is over 30%, and the annual economic loss is as high as 16 billion yuan. The disease problem has become an important factor restricting the healthy development of mariculture industry. [0003] For the occurrence of various diseases, chemotherapy represented by antibiot...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/195C12N15/31C12N15/63A61K39/02A61P31/04
Inventor 吴海珍张萌李小勇沈斌兵王启要张元兴
Owner EAST CHINA UNIV OF SCI & TECH
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