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

Application of koi Serum amyloid protein A5 or coding gene thereof in regulation of koi pathogenic bacteria infection resistance

A technology of serum starch and anti-pathogenic bacteria, applied in the field of genetic engineering, can solve the problems of less research and achieve the effect of improving the ability of anti-pathogenic bacteria infection

Pending Publication Date: 2021-11-30
BEIJING FISHERIES RES INST
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But for fish, there are few studies involving the function of SAA

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
  • Application of koi Serum amyloid protein A5 or coding gene thereof in regulation of koi pathogenic bacteria infection resistance
  • Application of koi Serum amyloid protein A5 or coding gene thereof in regulation of koi pathogenic bacteria infection resistance
  • Application of koi Serum amyloid protein A5 or coding gene thereof in regulation of koi pathogenic bacteria infection resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Obtaining of the Encoding Gene of Koi Serum Amyloid A5

[0036] In order to study the activation and up-regulation of immune-related genes in koi carp infected by pathogenic bacteria, this example performed transcriptome sequencing on the spleen of koi carp infected with Aeromonas victorii. According to the sequencing results, a significantly increased expression level was found. (about 189 times) differentially expressed genes. Further, according to homologous sequence alignment and protein structure simulation analysis, it is speculated in this embodiment that the protein encoded by the gene is Serum amyloid A-5 protein (koi serum amyloid A5).

[0037] In this embodiment, the cDNA of the koi spleen is further used as a template to amplify by PCR to obtain the open reading frame ORF of the koi Serum amyloidA-5 gene, which has a total of 372 nucleotides and encodes a protein consisting of 123 amino acids. Contains signal peptide. The amino acid sequence of K...

Embodiment 2

[0048] Example 2 Expression changes of koi carp Serum amyloid A-5 gene after Aeromonas verkis infection

[0049]Aeromonas veronii (A.v) was purchased from China General Microorganism Culture Collection Center (CGMCC, strain number: 1.927). After taking it out from the ultra-low temperature refrigerator, streak on the LB plate, and cultivate it in a 28°C incubator, recover the strain, pick a single colony and expand it in the LB liquid medium, collect the bacteria by centrifugation, resuspend with PBS, and adjust to 5 ×10 8 CFU / mL.

[0050] Randomly select 60 healthy koi carps (about 20 g) and divide them into 2 groups on average, the challenge group and the control group, with 3 parallel tanks in each group. Each koi in the challenge group was intraperitoneally injected with 100 μL of Aeromonas victorii suspension, and each koi in the control group was injected with 100 μL of PBS buffer intraperitoneally. Before infection (0h) and 6h, 12h, 24h, 48h, 96h, and 7d, 6 koi carp ...

Embodiment 3

[0052] Example 3 Verification of anti-pathogen infection ability of koi serum amyloid A5

[0053] 1. Construction of Koi Serum amyloid A-5 eukaryotic expression plasmid

[0054] (1) According to the mRNA sequence of the Koi Serum amyloid A-5 gene in the sequencing results, the amplification primers carrying KpnI and XhoI restriction sites were designed respectively:

[0055] F: 5'-GGGGTACCGGGATGGAGCTTATTCTTGCT-3',

[0056] R: 5'-CCCTCGAGTCAGTGGTGGTGGTGGTGGTGGTACTT-3'.

[0057] (2) Carry out PCR amplification with koi spleen cDNA as a template and primers F and R to obtain a cDNA fragment containing the ORF of the SAA-5 gene.

[0058] The PCR amplification reaction system (25 μL) contains 2.5 μL 10×Ex buffer, 2 μL dNTP, 0.3 μmol each of F and R primers, 1U Ex taq enzyme (Takara), 500 ng cDNA template;

[0059] The PCR reaction program was: 95°C for 5min; 92°C for 15s, 53°C for 25s, 72°C for 30s (10 cycles); 95°C for 15s, 60°C for 25s, 72°C for 30s (25 cycles); 72°C for 7min....

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 technical field of genetic engineering, and particularly relates to application of koi Serum amyloid protein A5 or a coding gene thereof in regulation of koi pathogenic bacteria infection resistance. A differential expression gene, namely a koi Serum amyloid A-5 gene, is obtained through research on koi infected with pathogenic bacteria, and the expression level of the gene in different tissues is remarkably changed after the koi is infected with the pathogenic bacteria. The koi Serum amyloid A-5 gene is overexpressed in a healthy koi body, the pathogenic bacteria infection resistance of the gene is remarkably improved, and the discovery has important significance in the field of resistance to pathogenic bacteria infection of koi.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the application of koi serum amyloid A5 or its coding gene in regulating koi anti-pathogen infection. Background technique [0002] Koi (Cyprinus carpio haematopterus) is an existing common ornamental fish. In the field of Koi culture, various diseases caused by pathogenic bacteria are a major problem in the process of Koi culture. In the prior art, the adjustment of the breeding environment, daily water quality management, feed management and other methods are usually used to avoid the disease of the koi carp by enhancing the resistance of the koi carp; or by raising the water temperature and using antibiotics after signs of disease appear for treatment. However, these methods are only effective against some pathogenic bacteria. [0003] Serum amyloid A (SAA) is an acute phase protein, secreted and synthesized by the liver, and involved in pathogen defense and immu...

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): C07K14/46C12N15/12C12N15/85A01K67/027
CPCC07K14/461C12N15/8509A01K67/0278A01K2207/15A01K2217/072A01K2227/40A01K2267/02Y02A40/81
Inventor 王晓雯朱华刘丽丽伍广涛
Owner BEIJING FISHERIES RES INST
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