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

Recombinant bovine nodule rash virus for expressing bovine herpesvirus I type gB gene and application of recombinant bovine nodule rash virus

A technology of bovine herpes virus, nodules, applied in the field of recombinant bovine nodular herpes virus

Active Publication Date: 2021-09-14
天康制药股份有限公司
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the vaccines of these schemes all need to be emulsified after inactivation, and can only prevent a single disease after immunization

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
  • Recombinant bovine nodule rash virus for expressing bovine herpesvirus I type gB gene and application of recombinant bovine nodule rash virus
  • Recombinant bovine nodule rash virus for expressing bovine herpesvirus I type gB gene and application of recombinant bovine nodule rash virus
  • Recombinant bovine nodule rash virus for expressing bovine herpesvirus I type gB gene and application of recombinant bovine nodule rash virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The preparation of embodiment 1 transfer vector plasmid

[0055] Construction of pSP72-p7.5-gB-TK1 plasmid

[0056] According to the gene sequence of P7.5 and TK1, a restriction site HindIII (AAGCT) was added at the 5' end of the P7.5 promoter, and a restriction site BamH I (GGATC) was added at the 3' end; at the 5' end of TK1 The enzyme cutting site SalII (CCGCGG) is added, and the enzyme cutting site SacII (CCGCGG) is added at the 3' end; P7.5 and TK1 are linked together and artificially synthesized.

[0057] The restriction site BamHI (GGATC) was added to the 5' end of the gB gene, and the restriction site SalII (CCGCGG) was added to the 3' end, which was artificially synthesized.

[0058] The vector pSP72 and P7.5-TK1 were double-enzyme-digested with HindIII and SacII respectively. After the enzyme-digestion was completed, the digestion buffer was removed with a gel recovery kit to obtain linear pSP72 and P7.5-TK1 with restriction sites. Ligate with T4 DNA ligase,...

Embodiment 2

[0068] The preparation of embodiment 2 recombinant bovine nodular rash virus

[0069] Spread LT cells (sheep testicular cells) on a 6-well plate. After the cells grow into a single layer, inoculate the LT cells on the 6-well plate with the attenuated bovine nodular rash strain at a multiplicity of infection (M.O.I.) of 0.1, at 37°C Adsorbed for 2 hours, the prepared recombinant transfer plasmid pB-TK1-gB-p7.5-P11-EGFP-TK2 was transfected by liposome, according to Lipofectamine TM 2000Transfection Reagent kit instruction manual for transfection respectively. Use 2-3 μg of pB-TK1-gB-p7.5-P11-EGFP-TK2 recombinant plasmid per well of cells,

[0070] The amount of liposome used is about 3 times that of DNA (mass volume ratio), and diluted with OPTI-MEM about 100-200 μL. After 48 hours of transfection, observe whether there is fluorescence under a fluorescence microscope, and observe the gradual development of cell lesions at the same time. When 90% of the cells were diseased and...

Embodiment 3

[0073] Domestication of embodiment 3 recombinant bovine nodular herpes virus

[0074] Vero adherent cells were cultured in serum-containing medium after resuscitation, and the amount of serum added was 8%. After 2-3 generations of continuous passage, the serum content began to decrease after the cell proliferation was stable, and 2% serum was reduced for each passage. When the serum concentration After it drops to 5%, add 2% yeast extract and 2% plant hydrolyzed protein to the medium. If the growth is slow, continue to maintain the original serum concentration until the serum content drops to 1%. After the cells grow stably, culture for 1-2 After passage, Vero adherent cells were directly transferred from square flasks to shake flasks for suspension culture. Using serum-free suspension medium, adding 1% yeast extract and 1% plant hydrolyzed protein to the medium, Vero suspension cells were cultured in shake flasks at 1.0×10 6 cells / mL is the initial seeding density, calculate...

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

PropertyMeasurementUnit
Titeraaaaaaaaaa
Login to View More

Abstract

The invention relates to the technical field of biology, and particularly provides a recombinant bovine nodule rash virus for expressing a bovine herpesvirus I type gB gene and application of the recombinant bovine nodule rash virus. The recombinant bovine nodule rash virus contains the bovine herpes virus I type gB gene as shown in SEQ ID NO.1. The recombinant bovine nodule rash virus has antigen epitopes of the two viruses, so that direct inoculation can be realized by using the virus as a live vector vaccine, operations such as inactivation and adjuvant emulsification are avoided, and the recombinant bovine nodule rash virus has good immunogenicity on bovine nodule rash virus and infectious bovine rhinotracheitis virus, and two prevention by one injection is achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant bovine nodular herpes virus expressing bovine herpes virus type I gB gene and application thereof. Background technique [0002] Bovine herpesvirus type Ⅰ (BHV-Ⅰ) belongs to the α-herpes virus and causes infectious bovine rhinotracheitis (IBR), which manifests as clinical symptoms such as respiratory disease, reproductive tract damage, abortion and even fatal systemic infection. There are at least 10 kinds of glycoproteins on the envelope surface of BHV-I, among which glycoprotein gB is an important molecule involved in the mechanism of BHV-I invading host cells and is necessary for virus replication. At the same time, gB is also one of the main antigenic proteins that stimulate the host's immune response, and can induce the host to produce high levels of humoral and cellular immune responses. [0003] Lumpy skin disease (LSD), also known as bovine nodular rash, bovine...

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): C12N7/01C12N15/38C12N15/63A61K39/275A61K39/265A61K9/19A61K47/42A61K47/26A61K47/10A61K47/18A61P31/20A61P31/22
CPCC12N7/00C07K14/005A61K39/12A61K9/19A61K47/42A61K47/26A61K47/10A61K47/183A61P31/20A61P31/22C12N2710/24021C12N2710/16722C12N2710/24034C12N2710/16734A61K2039/552A61K2039/70A61K2039/5156Y02A50/30
Inventor 贺笋李俊辉潘毅平赵毅程兰玲张伟唐慧芬张慧敏张志鹏
Owner 天康制药股份有限公司
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