Streptococcus suis vaccine recombinant protein GSE and preparation method and application thereof

A recombinant protein, Streptococcus suis technology, which is applied in the biological field to achieve the effect of good practicability and efficient preparation

Inactive Publication Date: 2021-02-26
TIANJIN AGRICULTURE COLLEGE
View PDF4 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have confirmed that these protective antigens have good immune protection, but no researchers have combined these thr

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
  • Streptococcus suis vaccine recombinant protein GSE and preparation method and application thereof
  • Streptococcus suis vaccine recombinant protein GSE and preparation method and application thereof
  • Streptococcus suis vaccine recombinant protein GSE and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0055] The preparation method of the above-mentioned recombinant protein comprises the following steps:

[0056] (1) B cell and helper T cell epitope screening;

[0057] (2) B cell and helper T cell epitope tandem gene design and plasmid construction;

[0058] (3) Prokaryotic expression and purification of the recombinant protein to obtain the recombinant protein GSE of the Streptococcus suis vaccine.

[0059] Preferably, the specific method of step (1) is: according to the amino acid sequence of Streptococcus suis glyceraldehyde triphosphate dehydrogenase (GAPDH), enolase (Enolase), DNA nuclease (SsnA), apply bioinformatics software , analyze the primary structure of the protein, exclude the transmembrane region and the signal peptide region during epitope screening, then predict the B cell and helper T cell epitope, and finally analyze the secondary structure of the protein to further verify the screened epitope Whether it is in the area of ​​antigenicity and hydrophilicit...

Embodiment 1

[0076] Antigen epitope screening:

[0077] 1. Protein primary structure analysis

[0078] The protein amino acid sequences of Streptococcus suis GAPDH, SsnA, and Enolase published by NCBI are respectively AKG39592.1, ABP90934.1, and ACS66679.1, and the sequence lengths are 336aa, 1059aa, and 435aa, respectively. According to the protein amino acid sequences of GAPDH, SsnA and enolase, the transmembrane region of the protein was analyzed by TMHMM tool. The SignalP tool was used to analyze the protein signal peptide region.

[0079] The results of analysis using the TMHMM tool showed that there was no transmembrane region in the GAPDH protein, and its 336 amino acids were all extracellular fragments (such as figure 1 shown). The SsnA protein has two transmembrane regions, the N-terminal amino acid 1-27 is an intracellular fragment, the 28-50 amino acid is a transmembrane fragment, the C-terminal amino acid 1054-1059 is an intracellular fragment, and the 1036-1053 amino acid i...

Embodiment 2

[0096] Recombinant protein design and plasmid construction:

[0097] The predicted B cell and helper T cell epitopes were spliced ​​sequentially in the order of GAPDH-SsnA-Enolase, and a dendritic cell targeting peptide sequence (sequence FYPSYHSTPQRP) was added to the N-terminus of the recombinant polypeptide. Each peptide segment Four glycine (GGGG) short peptides were used to connect them. After the concatenation is completed, the amino acid sequence of the recombinant protein is obtained, with a total of 373 amino acids, as shown in SEQ ID NO.1.

[0098] The amino acid sequence is translated into a nucleotide sequence, and then the obtained sequence is optimized according to the codon preference of E. coli, and a restriction site BamH1 is added to the N-terminal of the sequence, and a restriction site XhoI is added to the C-terminal of the sequence to finally obtain the recombinant The nucleotide sequence of the protein GSE has a total of 1131 nucleotides, as shown in SEQ...

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 a recombinant protein GSE used as a streptococcus suis vaccine. The amino acid sequence of the recombinant protein is SEQ ID NO. 1. Aiming at the problems that the cross protection effect between an inactivated vaccine and a low virulent vaccine of streptococcus suis is poor, and certain biosecurity exists, the recombinant protein designed on the basis of screening of streptococcus suis protective antigen epitopes can be used for preparing the streptococcus suis vaccine. The recombinant protein can be efficiently prepared under the safe condition and has good practicability.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a Streptococcus suis vaccine recombinant protein GSE and its preparation method and application. Background technique [0002] Streptococcus suis (Streptococcus suis) is a very important bacterial infectious disease pathogen for the pig industry. If an epidemic occurs, it may cause huge economic losses. Streptococcus suis can cause meningitis, arthritis, sepsis, Diseases such as endocarditis, encephalitis, miscarriage, multiple serositis, and bronchopneumonia have a mortality rate as high as 80%. Large-scale outbreaks occurred in my country in 1990 and 2005, which once again reminded people to pay more attention to the disease. [0003] Due to the large number of serotypes of Streptococcus suis, the inter-type cross-protection effect of inactivated vaccines is not good in the preventive stage of this disease. The current vaccine research is more inclined to the development of subun...

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): C12N9/02C12N9/88C12N9/22C12N15/62C12N15/70A61K39/09A61P31/04
CPCA61K39/092A61K2039/552A61P31/04C07K2319/33C07K2319/50C12N9/0008C12N9/22C12N9/88C12N15/70C12N2800/101C12Y102/01009C12Y402/01011
Inventor 赵瑞利潘晨浩金天明姜轩张欣李留安于恩远宋淇淇于晓雪
Owner TIANJIN AGRICULTURE COLLEGE
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