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SNP site related to stress resistance of lateolabrax japonicus and application of SNP site

A stress-resistant and site-based technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as germplasm degradation, lack of excellent species of perch, destruction of wild perch resources, etc. To achieve the effect of good immune performance

Active Publication Date: 2021-08-17
HUZHOU TEACHERS COLLEGE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, my country's perch industry is currently facing unprecedented challenges, mainly in the degradation of germplasm and the lack of good species of perch. The northern region still uses wild fish fry as seedlings, resulting in the destruction of wild perch resources, and artificially cultivated seedlings survive. The rate is extremely low; no targeted breeding has been carried out, scientific breeding techniques have not been established, and long-term inbreeding leads to reduced growth and disease resistance

Method used

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  • SNP site related to stress resistance of lateolabrax japonicus and application of SNP site
  • SNP site related to stress resistance of lateolabrax japonicus and application of SNP site
  • SNP site related to stress resistance of lateolabrax japonicus and application of SNP site

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Screening SNP sites

[0019] 1. Extraction of total RNA from sea bass:

[0020] Total RNA was extracted from the liver of sea bass according to the Trizol method.

[0021] 1) Tissue: Take 5mg of liver tissue and place it in a 1.5ml centrifuge tube, add 1ml Trizol to fully homogenate, and let stand at room temperature for 5 minutes;

[0022] 2) Add 0.2ml chloroform, shake for 15s, let stand for 2min; then centrifuge at 4°C, 12000g×15min, take the supernatant;

[0023] 3) Add 0.5ml of isopropanol to the supernatant, mix the liquid in the tube gently, and let it stand at room temperature for 10 minutes; then centrifuge at 4°C, 12000g×10min, and discard the supernatant.

[0024] 4) Add 1 ml of 75% ethanol to the precipitate, and gently wash the precipitate. 4°C, 7500g×5min, discard the supernatant, add appropriate amount of DEPC H after drying 2 O dissolved.

[0025] 2. Synthesize cDNA:

[0026] Using 2μg total RNA as a template, add 1μl 20μM Oligo d(T) 18...

Embodiment 2

[0045] Example 2: Homozygous individuals with genotype 264CC carry out infection experiments

[0046] Selected genotype 264CC Chinese perch individuals were selected for the injection infection experiment of Edwardsiella tarda, and the intraperitoneal injection concentration of each tail was 4.0×10 6 50 μL of CFU / mL bacterial solution; regularly observe and record the number of dead fish in each experimental group every day, and calculate the mortality rate of individuals with different genotypes.

[0047] The results showed that individuals with genotype 264GG had a higher incidence in injected infected individuals than individuals with the other two genotypes (Table 2).

[0048] Table 2: Injection infection incidence of three genotypes of Chinese perch

[0049] genotype number of cases Morbidity Fatalities lethal dose rate CC 2 / 18 11％ 0 0％ CG 9 / 30 30％ 3 10％ GG 23 / 30 77％ 19 63％

[0050] Note: Indicates survival, number of deat...

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Abstract

The invention provides an SNP (Single Nucleotide Polymorphism) site related to stress resistance of lateolabrax japonicus. The site is positioned at the 264th site of a nucleotide fragment with a sequence of SEQ ID NO: 1, the basic group of the site is G or C, and the coding protein is replaced by Asn from Lys. The SNP site provided by the invention is used for breeding a lateolabrax japonicus individual with better stress resistance. By analyzing the genes of the Chinese lateolabrax japonicus with different disease resistance properties, it is found that the stress resistance related SNP sites exist on the CD4-1 gene of the Chinese lateolabrax japonicus, and a homozygous individual with the genotype of 264CC has better immune performance. Therefore, the genotype individual can be selected as a parent for genetic breeding in production.

Description

technical field [0001] The invention belongs to the technical field of fish genetics and breeding, and specifically relates to a SNP site related to the stress resistance of Chinese sea bass and an application thereof. Background technique [0002] Lateolabrax maculatus belongs to the order Perciformes, Lateolabrax family, and the genus Lateolabrax. It is distributed in my country, Japan, and the coastal areas and estuaries of the Korean Peninsula. It breeds in seawater and can grow in various types of waters. Lateolabrax maculatus is an important seafood in my country. Economic fish can survive and grow in seawater, brackish water, freshwater or estuary areas. [0003] my country's perch breeding area is relatively wide, mainly concentrated in the coastal areas of Guangdong, Shandong, Fujian, Zhejiang, Guangxi and other provinces. Moreover, the demand for sea bass fry in inland aquaculture waters is also increasing. But at present, my country's perch industry is facing unp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/124
Inventor 朱俊杰陆国富叶金云
Owner HUZHOU TEACHERS COLLEGE
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