Bacterium-delivered double cross protection vaccine

A cross-protection and vaccine technology, applied in the field of vaccinology, can solve problems such as cross-protection vaccines, and achieve the effect of protecting infection and simple preparation

Inactive Publication Date: 2012-09-12
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there have been many reports of single vaccines against Edwardsiella tarda and Streptococcus iniae respectively in the world, but there is no cross-protective vaccine against these two pathogens at the same time

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
  • Bacterium-delivered double cross protection vaccine
  • Bacterium-delivered double cross protection vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] 1) Construction of plasmid pEADnaK: Edwardsiella tarda TX1 was used as a template, and F1 / R1 was used as primers for PCR amplification. The PCR conditions were: 94°C for 60s to pre-denature the template DNA, then 94°C for 40s, 51°C for 60s, and 72°C 60s, after 5 cycles, change to 94°C for 40s, 61°C for 60s, 72°C for 60s, and after 25 cycles, extend the reaction at 72°C for 10min. The PCR product was purified with the "Tiangen Biochemical Technology (Beijing) Co., Ltd." DNA product purification kit, and then connected to the carrier pEASY-E2 (purchased from "Beijing Quanshijin Biotechnology Co., Ltd.") at room temperature for 2-4 hours. After the mixture was transformed into Escherichia coli DH5α, it was cultured on LB solid medium containing 100 ug / ml anka penicillin (Ap) for 18 hours, a transformant was picked, and the plasmid was extracted, which was pEADnaK.

[0013] The LB composition is calculated by weight percentage: 1.0% peptone, 0.5% yeast powder, 1.0% sodium c...

Embodiment 2

[0021] Construction of the vaccine: Transform the plasmid pDKS10 into Escherichia coli DH5 according to the Hanahan method (Sambrook and Russell: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press 2001) to obtain strain DH5α / pDKS10; / ml) on the LB medium for 18-24 hours, the plasmid was extracted, and the plasmid was found to be plasmid pDKS10 through sequencing, indicating that the bacterial strain DH5α / pDKS10 did contain pDKS10. Strain DH5α / pDKS10 is a vaccine strain with cross-protection against Edwardsiella lentus and Streptococcus iniae. As a control, the plasmid pBT3 was transformed into Escherichia coli DH5 in the same way to obtain strain DH5α / pBT3, which was the control strain.

Embodiment 3

[0023] Application of the vaccine: Step 1) Preparation of the vaccine preparation solution. The vaccine strain DH5α / pDKS10 obtained above and the control strain DH5α / pBT3 were cultured in LB liquid medium to OD 600 0.8-1, then centrifuge the culture solution (5000g, 4°C, 10min), collect the bacteria, and suspend them in PBS to a final concentration of 1×10 8 cfu / ml is the vaccine preparation solution and control solution.

[0024] Step 2) Vaccination. 120 flounder (each weighing about 12.2g) were randomly divided into 4 groups, 30 in each group. These 4 groups are named A, B, C and D respectively. Each fish in groups A and C was injected intraperitoneally with 100 ul of the vaccine preparation solution in step 1) above, and each fish in groups B and D (control group) was injected intraperitoneally with 100 ul of the control solution in step 1) above.

[0025] Step 3) Preparation of Edwardsiella tarda and Streptococcus iniae suspensions. Cultivate Edwardsiella tarda TX1 an...

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 present invention relates to a field of vaccinology, and especially relates to a bacteria-delivered cross protection vaccine against Edwardsiella tarda and Streptococcus iniae. The cross protection vaccine is a recombinant strain obtained by plasmid [rho]DKS10 transforming escherichia coli DH5[alpha]. The vaccine obtained in the present invention has 83% and 75% protection efficiency respectively for Edwardsiella tarda and Streptococcus iniae infection.

Description

technical field [0001] The invention relates to the field of vaccinology, in particular to a cross-protection vaccine of Edwardsiella tarda and Streptococcus iniae transmitted by bacteria. Background technique [0002] Edwardsiella tarda and Streptococcus iniae are Gram-negative and Gram-positive bacteria, respectively, both of which are important pathogens of farmed fish, and their host range covers a variety of marine and freshwater Fish, including flounder, turbot, tilapia, redfish, etc. In my country, outbreaks of diseases caused by Edwardsiella tarda and Streptococcus iniae have caused serious economic losses to a variety of cultured fish industries. In addition to fish, Edwardsiella tarda and Streptococcus iniae are zoonotic pathogens that can infect humans and other terrestrial animals. At present, there have been many reports of single vaccines against Edwardsiella tarda and Streptococcus iniae respectively in the world, but there is no cross-protective vaccine tha...

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): A61K39/116C12N1/21A61P31/04A61K39/02A61K39/09
Inventor 孙黎胡永华党伟
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap