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Genetic marker-related with immunity traits of hemorragic disease of grass carp

A grass carp hemorrhagic disease and genetic marker technology is applied in the field of genetic markers related to the immune traits of grass carp hemorrhagic disease, which can solve the problems of time-consuming and labor-consuming, and achieve the effects of accurate detection, high throughput and reliable results

Active Publication Date: 2016-12-28
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional breeding method is time-consuming and labor-intensive, so the breeding method based on genetic markers is gradually adopted by researchers.

Method used

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  • Genetic marker-related with immunity traits of hemorragic disease of grass carp
  • Genetic marker-related with immunity traits of hemorragic disease of grass carp
  • Genetic marker-related with immunity traits of hemorragic disease of grass carp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Transcriptome sequencing of grass carp spleen, head kidney and monoclonal CIK cells

[0048] (1) Infection test

[0049] The aquarium used in this example was soaked in 0.05% potassium permanganate solution for 24 hours, then cleaned, injected with clear water, installed the oxygenation pump and heating rod, set the temperature to 28°C-30°C, and started the oxygenation pump Then prepare the grass carp for temporary breeding test. Before Grass carp reovirus (GCRV) infection, the grass carp were randomly divided into five groups (60 / group) and kept in the prepared aquarium, and the palatable particles were fed twice a day at a fixed point, time, and quantity. Feed, and clean up residual bait and metabolic waste in time. For about a week, prepare for the virus infection test after the daily physiological activities of the fish school are normal. The stock solution of GCRV-097 strain was donated by the Institute of Hydrobiology, Chinese Academy of Sciences. T...

Embodiment 2

[0074] Example 2: C7N1 Gene Verification and Association Analysis

[0075] (1) DNA extraction of grass carp spleen tissue

[0076] Genomic DNA was extracted by conventional chloroform method. The specific operation method is as follows:

[0077]1) Take 0.05g of tissue (the size of mung bean) and put it in a 1.5mL EP tube, add 200μL of tissue extract, and grind it thoroughly. Add 400 μL of tissue extract and 60 μL of 1 mg / mL proteinase K, put in a 55°C water bath (for about 3 hours) or overnight at 37°C, and digest until the liquid is clear;

[0078] 2) Add 600 μL Tris-saturated phenol, mix thoroughly for 10 minutes, and centrifuge at 12,000 rpm for 10 minutes;

[0079] 3) Transfer the supernatant to a new EP tube, add 500 μL chloroform for extraction, mix thoroughly for 10 minutes, and centrifuge at 12,000 rpm for 10 minutes;

[0080] 4) Pipette the supernatant into a new EP tube, add 2 times the volume of ice ethanol (-20°C), precipitate DNA at -20°C for at least 30 minut...

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Abstract

The invention belongs to the technical field of fish genetic marker screening and preparation, and more specifically relates to a genetic marker-related with immunity traits of hemorragic disease of grass carp. The genetic marker is obtained by screening C7N1 gene. The genetic marker is obtained by steps of grass carp individual level resistant colony and sensitive colony transcriptome sequencing, cell level resistance and sensitive cell transcriptome sequencing, correlation analysis, and infection validation, the genetic marker is shown as SEQ ID NO:1, a base substitution mutation of C7N1 gene at 2145th bit base, and the change of 715th amino acid of grass carp C7N1 gene is generated. The grass carp reovirus infection test shows that heterozygous gene-type grass carp mortality is obviously lower than the homozygotic-type grass carp mortality, and shows that the grass carp C7N1 gene base mutation is related with the grass carp hemorragic disease property. The novel genetic marker resource can be provided for seed selection for immunity traits of hemorragic disease of grass carp.

Description

technical field [0001] The invention belongs to the technical field of fish genetic marker screening and preparation, and in particular relates to a genetic marker related to immune traits of grass carp hemorrhagic disease. The genetic markers are obtained from the C7N1 gene. The present invention also relates to the application of the genetic marker in the association analysis of the genetic marker of the immune character of grass carp hemorrhagic disease. Background technique [0002] Grass carp (Ctenopharyngodon idella, English name grass carp) is an important freshwater aquaculture fish in my country, and its output accounts for about 18% of the total freshwater aquaculture output (Rao YL et al. 2015. Insights into the antiviral immunity against grass carp (Ctenopharyngodon idella) reovirus (GCRV ) in grasscarp. J Immunol Res, 670437). Grass carp haemorrhagic disease is the most serious disease of grass carp species. The disease has a long onset season, a wide range of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 苏建国廖志伟万全元尚雪萤
Owner HUAZHONG AGRI UNIV
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