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SNP molecular marker related to siniperca chuatsi infectious spleen and kidney necrosis disease resistance, detection method and application of SNP molecular marker

A molecular marker, necrosis technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, recombinant DNA technology, etc., to achieve the effect of improving economic value

Active Publication Date: 2021-06-29
SUZHOU UNIV
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there are few researches on fish IFN-α3 gene related to disease resistance, so it is of great research significance to obtain the full-length gDNA sequence of mandarin fish IFN-α3 and explore the correlation of this gene with disease resistance

Method used

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  • SNP molecular marker related to siniperca chuatsi infectious spleen and kidney necrosis disease resistance, detection method and application of SNP molecular marker
  • SNP molecular marker related to siniperca chuatsi infectious spleen and kidney necrosis disease resistance, detection method and application of SNP molecular marker
  • SNP molecular marker related to siniperca chuatsi infectious spleen and kidney necrosis disease resistance, detection method and application of SNP molecular marker

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Experimental program
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Embodiment 1

[0039] Embodiment 1 Siniperca IFN-α3 gene full-length sequence amplification

[0040] 1. Amplification of intron of mandarin fish IFN-α3 gene

[0041] Design specific primers to amplify the intron of the IFN-α3 gene with reference to the complete fragment of the known mandarin fish IFN-α3 cDNA (see Table 1), and use the mandarin fish genome DNA as a template to carry out fragmented PCR amplification of the gene. The amplification procedure is shown in Table 2. The PCR products were sequenced. The obtained sequences were spliced ​​using manual alignment and DNAMAN software to obtain the full length of the mandarin fish IFN-α3 gDNA gene, as shown in SEQ ID NO.1. The gel electrophoresis results after adding the primers used for the intron to carry out the PCR amplification of the mandarin fish IFN-α3 gene are as follows: figure 1 As shown, where a is the electrophoresis pattern of IFN-a3-4F, 4R amplification products, b is the electrophoresis pattern of IFN-a3-3F, 3R amplifica...

Embodiment 2

[0057] Example 2 Detection results of disease-resistant SNP markers based on mandarin fish IFN-α3 gene

[0058] The mandarin fish of the same group were all cultured under the same feeding conditions. After about two months of breeding, 100 fish were randomly selected from the cultured group for the challenge test. Each fish in each group was intraperitoneally injected with infectious spleen and kidney necrosis virus. (ISKNV, also known as iridescent virus)10 3.68 TCID50 / ml, 200μl, continuous observation for 10 days, the symptoms observed within 10 days are the same as the symptoms of ISKNV infection in the natural environment, the infected fish swim slowly on the water surface or even directly float on the water surface, the body surface is not damaged, and the body color is white , the gills of the fish showed ischemia-like whitishness, and the dissection found: there was more ascites in the abdominal cavity, hyperemia in the liver, stomach wall, and intestinal wall, and yel...

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Abstract

The invention relates to an SNP molecular marker located on a siniperca chuatsi IFN-alpha 3 gene and related to siniperca chuatsi infectious spleen and kidney necrosis disease resistance. According to the invention, on the basis of known cDNA, cloning is further carried out, an intron and a gDNA sequence of the siniperca chuatsi IFN-alpha 3 gene are obtained, a primer is designed according to the IFN-alpha 3 gDNA sequence, through product amplification and sequencing comparison, SNP sites related to the siniperca chuatsi germplasm disease resistance or susceptibility character are determined, the disease resistance or susceptibility siniperca chuatsi germplasm can be rapidly screened out, and the method is a novel method for disease resistance germplasm detection. According to the method, the problem of lack of the fish IFN-alpha 3 gene is solved; and meanwhile, a theoretical basis and an operation method are provided for disease resistance germplasm breeding by utilizing the heredity of the SNP sites and the correlation between the SNP sites and disease resistance characters, so that ISNKNV disease resistance populations are accurately bred, the breeding work efficiency is improved, diseases are reduced, economic losses are reduced, and the breeding economic value is increased.

Description

technical field [0001] The invention relates to the fields of aquaculture and biotechnology, in particular to a SNP molecular marker related to the resistance of mandarin fish infectious spleen and kidney necrosis, a detection method and an application thereof. Background technique [0002] Mandarin fish (Sinipercachuatsi) belongs to Perciformes, Siniperidae, and Siniperca genus. Its meat is delicious, rich in high protein, various vitamins and trace elements, and it is an important economic fish species in my country. However, due to the sharp decline of wild original germplasm resources, weak disease resistance, and germplasm degradation, infectious spleen and kidney necrosis (ISKNV) has become the most serious fulminant viral disease in the culture of mandarin fish, causing huge losses. Economic losses. [0003] Traditional breeding techniques mainly rely on naked eye observation, but this method of breeding is difficult, time-consuming and labor-intensive, and is prone ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156Y02A40/81C12Q1/6869C12Q1/701
Inventor 黄鹤忠薛源陆仲逸刘苗苗
Owner SUZHOU UNIV
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