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

Expression of vibrio alginolyticus outer membrane protein VA0760 and application thereof as vaccine component

A technology of outer membrane protein and vibrio alginolyticus, applied in the fields of application, medical preparations containing active ingredients, drug combinations, etc.

Inactive Publication Date: 2009-03-18
SUN YAT SEN UNIV
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, before the present invention was published, there was no publication or report on the immune regulation function of Vibrio alginolyticus outer membrane protein Pal (VA1061) mentioned in this patent application

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
  • Expression of vibrio alginolyticus outer membrane protein VA0760 and application thereof as vaccine component
  • Expression of vibrio alginolyticus outer membrane protein VA0760 and application thereof as vaccine component
  • Expression of vibrio alginolyticus outer membrane protein VA0760 and application thereof as vaccine component

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Cloning and identification of outer membrane protein Pal(VA1061) gene of Vibrio alginolyticus

[0014] 1. Amplification of Vibrio alginolyticus Pal (VA1061) gene

[0015] Since the full sequence of the Vibrio alginolyticus gene has not been determined, there is no report on the Pal (VA1061) gene sequence of this bacterium. In view of the complete sequence determination of Vibrio parahaemolyticus, which was published in the database GenBank (BA000031) in March 2006, we designed the Pal (VA1061) gene primers according to the Pal (VA1061) sequence published on the database for the lysolytic PCR amplification of Vibrio algae Pal (VA1061). The designed primers are: Upstream primer: 5'-ATA GGATCC ATGAAAAAACTAGCAGCGGT-3'; downstream primer: 5'-CGC AAGCTT TTATTGCTGAACTTGGTA-3'; the horizontal line shows the restriction site, the upstream primer introduces the BamHI restriction site, and the downstream primer introduces the EcoRI restriction site. Use bacterial genome DNA ...

Embodiment 2

[0022] Expression of Pal(VA1061) protein from Vibrio alginolyticus

[0023] 1 Predicted amino acid sequence

[0024] The gene Pal (VA1061) is a medium-length fragment, and it is analyzed by DNAssist software to obtain its complete open reading frame ORF

[0025] 2 Prokaryotic expression of recombinants

[0026] Pick a single colony of the recombinant plasmid and inoculate it into 1ml of LB liquid medium supplemented with kanamycin (50mg / ml) at a ratio of 1:500, culture overnight at 37°C until saturated, and inoculate the saturated culture to 5mL at an inoculum size of 1:100 In LB medium, culture at 37°C for about 2h to OD 600 It is about 0.6 or so. IPTG was added to the culture to a final concentration of 1 mmol / L, and culture was continued at 37° C. for 3 h. The induced culture was taken out and centrifuged at room temperature for 1 min at high speed to collect the bacteria. At the same time, a control group was set up, and the pellet was resuspended in 100 μL of 2×SDS gel...

Embodiment 3

[0028] 1. Purification of Vibrio alginolyticus Pal (VA1061) protein

[0029] The recombinant plasmid was picked and transformed into Escherichia coli BL21(DE3), spread in LB medium containing 100 μg / mL kanamycin, and cultured overnight at 37°C. Immediately pick a single colony, add 5mL LB solution and shake overnight at 37°C, transfer to 200mL liquid LB medium containing antibiotics at a ratio of 1:100, culture at 37°C for about 2.5-3h, OD 600 When = 0.6, add IPTG (final concentration: 1 mmol / L) to the bottle for induction, shake at 200 rpm / min at 30°C for 3 hours. Centrifuge at 6000g for 10 minutes at 4°C to collect the cells, wash the cells twice with normal saline, and weigh the wet weight of the cells.

[0030] Wash 3 times with buffer solution (50mmol / L Tris HCL, 1mmol / L EDTA, pH 8.0), sonicate the cells, centrifuge at 6000rpm / min for 5min, discard the precipitate; centrifuge at 12000rpm / min for 10min, and recover the precipitate. Then the inclusion body was buffered wi...

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 relates to the expression of a recombined outer membrane protein Pal (VA1061) of Vibrio alginolyticus and the action function of a vaccine component. Through cloning, expressing and purifying genes of the recombined outer membrane protein Pal (VA1061) of the Vibrio alginolyticus, a purified product can obviously improve the resistance of the fish to pathogenic bacteria such as Vibrio alginolyticus, pseudomonas fluorescens. As a target candidate of a subunit vaccine, the recombined outer membrane protein VA0760 of the Vibrio alginolyticus has great significance on the immunological prevention and treatment of controlling Vibrio infection.

Description

technical field [0001] The invention relates to the immune regulation function of an outer membrane protein (outermembrane protein, OM protein) Pal (VA1061) expressed by Vibrio alginolyticus. Background technique [0002] Vibrio alginolyticus is widely distributed in seawater and estuaries all over the world, and the number ranks first among vibrio alginolyticus. For the pathogenic microorganisms of Vibrio alginolyticus, the focus of research is immunological analysis, to clarify its pathogenic mechanism and find effective The prevention and treatment methods mainly include drugs, namely antibiotics and immune prevention and treatment. Although antibiotics have made great progress in the initial use, however, with the widespread use of antibiotics, a series of related problems have also emerged. On the one hand The irrational use of antibiotics increases the emergence of antibiotic-resistant bacteria and bacterial infections; on the other hand, artificially increasing the do...

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): C07K14/28C12N15/31A61K39/106A61P37/04
Inventor 彭宣宪熊莜鹏李惠
Owner SUN YAT SEN UNIV
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