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Molecular marker related to bacterial septicemia resistance character of silurus meridionalis and application of molecular marker

A technology of molecular markers and sepsis, which is applied in the direction of climate change adaptation, microbial measurement/inspection, biochemical equipment and methods, etc., to achieve the effect of promoting sustainable and healthy development and shortening the intergenerational interval

Pending Publication Date: 2022-05-31
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the molecular markers related to the resistance of aquatic animals to Aeromonas hydrophila infection, and most of them focus on the detection of Aeromonas hydrophila and the preparation of corresponding antibacterial vaccines.
However, there is no report on the molecular markers related to the disease resistance of the catfish.

Method used

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  • Molecular marker related to bacterial septicemia resistance character of silurus meridionalis and application of molecular marker
  • Molecular marker related to bacterial septicemia resistance character of silurus meridionalis and application of molecular marker
  • Molecular marker related to bacterial septicemia resistance character of silurus meridionalis and application of molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The processing of embodiment 1 Su's round belly catfish

[0031] The used catfish samples of the present invention are all taken from Fangcun Market, Guangzhou City, Guangdong Province. The weight of the catfish as samples is about 100g, and 500 tails are taken altogether.

[0032] After salvaged from the culture tanks, the samples of the catfish were raised temporarily until they were stable, and then injected with Aeromonas hydrophila to artificially induce infection. Among them, the injection concentration of Aeromonas hydrophila was 1×10 7 cfu / mL, the injection volume is 0.2 mL, and the water temperature is kept at 27-29 °C to continue culturing and observing. Collect the catfish that died first within 48 hours as the susceptible group; collect the catfish that had no symptoms of infection after 5 days as the resistant group; among them, 121 catfish were collected in the susceptible group For belly catfish, 124 were collected in the disease-resistant group.

[00...

Embodiment 2

[0034] Example 2 Cloning of CDS region of interleukin 4 gene

[0035] The present invention refers to the interleukin-4 (IL-4R) gene sequence published in the GenBank database, and utilizes Primer Premier 5 software to design primers for amplifying the CDS region of the interleukin-4 gene, respectively Named IL-4R-F and IL-4R-R. See Table 1 for primer information:

[0036] Table 1 Primers required for cloning of IL-4R gene CDS region and SNP screening

[0037]

[0038] From the susceptibility group preserved in embodiment 1 and the disease-resistant group Su Shi's round belly catfish, each randomly selects 10 Su Shi's round belly catfish samples and extracts genomic DNA, and the extracted genomic DNA is detected its concentration by ultraviolet spectrophotometer, Diluted to 100ng / μL for PCR amplification of CDS region of IL-4R gene.

[0039] The CDS region of the IL-4R gene was amplified using the primers shown in Table 1, using the diluted C. schneidi genomic DNA as a t...

Embodiment 3

[0045] Example 3 SNP site screening and typing

[0046] The software (DNAstar) was used to compare and analyze the sequenced 20 sequences of Catfish schneidi and the reference sequence (NCBI ReferenceSequence: CM018563.1). The ratio of different bases at the same site was greater than 1 / 3 as the SNP site, and one SNP site was initially obtained, which was named SNP1 G>A. The SNP1 site is located in the exon region of the CDS region of the IL-4R gene of the catfish sioglii, and the mutation of the SNP1 site is a non-synonymous mutation, as shown in Table 3.

[0047] Table 3 Statistics of single nucleotide polymorphism (SNP) sites in CDS region of IL-4R gene

[0048]

[0049] The invention determines the genotype of the SNP site by counting the bases of the SNP site of the sample and analyzing the sequencing peak. SNP1 C>T site sequencing information (three genotypes) such as figure 1 shown.

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Abstract

The invention discloses a molecular marker related to a bacterial septicemia resistant character of clarias fuscus. The molecular marker is located in an exon region of a CDS region of an interleukin 4 gene of the clarias suddenbergii, is related to antibacterial septicemia traits of the clarias suddenbergii, and can be applied to disease-resistant molecular marker-assisted breeding of the clarias suddenbergii, so that clarias suddenbergii individuals and families with relatively high disease resistance are cultured, and the breeding efficiency of the clarias suddenbergii is improved. And sustainable and healthy development of the silurus meridionalis breeding industry is promoted.

Description

technical field [0001] The invention belongs to the technical field of fish molecular marker assisted breeding and selection. More specifically, it relates to molecular markers related to anti-bacterial sepsis traits of catfish and their application. Background technique [0002] Pangasianodon hypophthalmus, also known as freshwater tiger shark, tiger head shark or pangasius, belongs to the Siluriformes, belongs to the genus Miscanthus, is a scaleless fish, and is an important freshwater aquaculture species in Southeast Asian countries. Sushi's round-bellied catfish has the characteristics of fast growth, low oxygen resistance, and no intermuscular spines, and is favored by the market. In recent years, with the continuous growth of the consumption demand of Sushi's round belly catfish in my country, the domestic breeding scale is also expanding day by day. However, with the improvement of the degree of intensive breeding, the degradation of economic traits caused by inbree...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156Y02A40/81
Inventor 刘文生阮灼豪蒋亮森卢志强黎遗富罗媛媛张细权
Owner SOUTH CHINA AGRI UNIV
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