Unlock instant, AI-driven research and patent intelligence for your innovation.

High-density fermentation supplemented culture medium and fermentation technology of African swine fever genetic engineering vaccines

A genetic engineering vaccine, high-density fermentation technology, applied in the field of African swine fever genetic engineering vaccine high-density fermentation feed medium and fermentation technology, can solve the problem of low expression level and achieve the effect of increasing production

Active Publication Date: 2020-06-09
河南省生物工程技术研究中心 +1
View PDF23 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The invention provides a high-density fermented feed medium for African swine fever genetic engineering vaccine. In the process of fermentation and cultivation, by adding the feed medium into the basic medium, the problem of using Escherichia coli to express African swine fever genetic engineering can be solved. The problem of low expression level in vaccines

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
  • High-density fermentation supplemented culture medium and fermentation technology of African swine fever genetic engineering vaccines
  • High-density fermentation supplemented culture medium and fermentation technology of African swine fever genetic engineering vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Designing the amino acid sequence of African swine fever genetically engineered vaccine

[0027] Wherein, the African swine fever genetically engineered vaccine (fusion protein) includes at least the following amino acid fragments: at least one neutralizing antigenic epitope selected from p30 protein, at least one neutralizing antigenic epitope selected from p54 protein, and at least one neutralizing antigenic epitope selected from p72 At least one neutralizing antigenic epitope of the protein (the corresponding amino acid sequence can be the corresponding sequence listed in Chinese patent documents CN201911062920.5, CN201911052111.6 or CN201911052128.1).

[0028] In addition to the above-mentioned essential amino acid fragments, the African swine fever genetically engineered vaccine may also include T cell epitope peptides, such as p30 protein T cell epitope peptides, T lymphocyte activation epitope peptides (including but not limited to influenza virus NP pro...

Embodiment 2

[0032] Example 2 Construction of recombinant Escherichia coli

[0033] Submit the amino acid sequence of the African swine fever genetically engineered vaccine to be expressed to Sangon Bio (Shanghai) Co., Ltd., and the company will synthesize the nucleotide sequence encoding the African swine fever genetically engineered vaccine to be expressed according to the provided amino acid sequence (gene of interest), and the synthesized nucleotide sequence was cloned into a suitable plasmid vector pET28a. The plasmid containing the target gene was obtained and transferred to BL21 (DE3) to obtain recombinant Escherichia coli and placed in a -70°C strain tube (containing 75% glycerol) for future use.

[0034] Among them, the amino acid sequence of the African swine fever genetically engineered vaccine is:

[0035] TKPRKK MDSEFFQPVYPRHYGECLSPVTTPSFFSTHMYTIL GGGSGGG RRNIRFKPWFIPGVINEISLT NNELYINNLFVTPEIHNLFVKRVRFSLIRVHKTQ GGGSGGG NETNECTSSFETLFEQEPSSE KKTYQRTRALVGGGSGGGG

Embodiment 3

[0036] Example 3 High Density Fermentation of Recombinant Escherichia coli

[0037] 1.1 Recombinant E. coli culture

[0038] It is mainly divided into four stages: strain activation, seed culture, shake flask culture and fermenter culture.

[0039] 1.1.1 Activation of strains

[0040] Recombinant Escherichia coli strains are stored in -70°C strain tubes (containing 75% glycerol), and due to their low activity, they need to be transferred to slant medium and cultured overnight in a constant temperature incubator (37°C). Then, under the condition of 37°C and 225r / min, the strains were activated for 10 hours by shaking the flask twice, and then inoculated on a plate containing kanamycin and stored at 4°C.

[0041] 1.1.2 Seed Cultivation

[0042] Pick a single colony on the plate containing kanamycin with an inoculating loop, and transfer the strain to a sterilized 50 mL seed medium (LB liquid medium, each liter containing 10 g NaCl, 10 g peptone, 5g yeast powder, 0.02M pH7.4P...

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 microorganism fermentation, and particularly relates to a high-density fermentation supplemented culture medium and fermentation technology of an African swine fever genetic engineering vaccines. The high-density fermentation supplemented culture medium of African swine fever genetic engineering vaccines comprises 40-280g / L glucose, 50-300g / L glycine, 20g / L MgSO4 and 20g / L (NH4)2SO4. The amino acid sequence of the African swine fever genetic engineering vaccines comprises the following fragments of one or more neutralization antigenic epitopes selected from p72 protein, one or more neutralization antigenic epitopes selected from p54 protein and one or more neutralization antigenic epitopes selected from p30 protein. Through the adoption of the fermentation technology disclosed by the invention, the yield of the African swine fever genetic engineering vaccines can be notably increased.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a high-density fermentation feed medium and a fermentation process for an African swine fever genetic engineering vaccine. Background technique [0002] Genetically engineered subunit vaccines use DNA recombination and DNA sequence synthesis of polypeptides to produce vaccines that contain only protective proteins and epitopes identified from pathogens. Usually, the first step in its production is to identify specific antigenic DNA fragments with immunogenicity in the pathogen, and the second step is to select a suitable vector for expression and amplification. At present, Escherichia coli is commonly used as an expression system for prokaryotic genes in production. Compared with conventional vaccines, subunit vaccines have the following advantages: greatly reduce the number of antigens while retaining specificity, reducing side reactions caused by irr...

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): C12N1/21C12P21/02C12R1/19
CPCC12P21/02C12N1/20C07K14/005C12N2710/12022Y02A50/30
Inventor 王云龙李玉林毕胜利伊瑶范雪亭张怡青程蕾王继创王国强
Owner 河南省生物工程技术研究中心