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SNP site related to rapid growth of large-sized takifugu rubripes fries and application of SNP site

A redfin puffer and site technology, applied in the field of fish genetic selection and breeding, can solve the problems of natural resource decline, germplasm degradation, etc.

Active Publication Date: 2020-10-20
DALIAN OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the amount of natural resources of pufferfish has declined seriously. Although the scale of breeding has gradually increased, germplasm degradation has occurred in the process of artificial breeding.

Method used

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  • SNP site related to rapid growth of large-sized takifugu rubripes fries and application of SNP site
  • SNP site related to rapid growth of large-sized takifugu rubripes fries and application of SNP site
  • SNP site related to rapid growth of large-sized takifugu rubripes fries and application of SNP site

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Screening SNP sites

[0024] The screening steps of the SNP site of the present invention are as follows:

[0025] a) Extraction of the redfin fugu genome: Genomic DNA was extracted from the redfin fugu muscle by the phenol-imitation extraction method. Take 100mg of its muscle and put it into a 1.5ml centrifuge tube, and use ophthalmic scissors to cut it into pieces as much as possible, add 500ul extract and 3μl proteinase K, and mix well; after mixing, put the centrifuge tube into a 55°C water bath for water bath Heat and turn over every 15 minutes. After the sample is cracked into a clear viscous liquid, take it out and add an equal volume of saturated phenol, and mix gently for 10 minutes. Centrifuge at 12000rpm for 10min; take the supernatant, add an equal volume of phenol / chloroform (1:1) mixture and mix gently for 10 minutes, put it in a centrifuge, and centrifuge at 12000rpm for 10min; take the supernatant, add an equal volume of Chloroform was mixed ...

Embodiment 2

[0032] Example 2: The PCR-SSCP method proves the correlation between SNP loci and growth traits

[0033] In order to determine the SNP site associated with the growth traits of the red fin puffer fragment whose sequence is SEQ ID NO: 1, the PCR-SSCP method detection is carried out, including the following steps:

[0034] a) Acquisition of red fin puffer samples: All the experimental sample fish used were collected from Dalian Fugu Aquaculture Co., Ltd., and were grown into randomly selected 102-day-old red fin puffer under the same breeding management and nutritional conditions There are 296 individual fry.

[0035] b) Data collection and genomic DNA extraction: the phenotype values ​​of body weight and body length of 296 individuals were measured and recorded, and muscle tissue was collected for genomic DNA extraction.

[0036] c) PCR reaction and sequencing. The reaction system is 25 μl: 10×buffer 2.5 μl, dNTP 2 μl, F primer (SEQ ID NO: 2) 1 μl, R primer (SEQ ID NO: 3) 1 μl,...

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Abstract

The invention provides an SNP site related to rapid growth of takifugu rubripes and a fry screening application capable of reducing fry selection cost and guranteeing good filial generation characters. The SNP site is located at the 162nd site of a nucleotide sequence as shown in SEQ ID NO: 1, wherein the base of the SNP site is C or G. The SNP site provided by the invention can be used for breeding takifugu rubripes individuals with rapid growth potential. According to the invention, through analysis of the correlation between the allelic SNP sit genotype and the takifugu rubripes growth characters, the SNP site related to the growth characters is found to exists at the 162 base of the nucleotide sequence, as shown in the SEQ ID NO: 1, of the takifugu rubripes, wherein the values of the weight and the body length of individuals with a CC homozygous genotype are significantly higher than the phenotype values (p (0.05)) of the growth characters of the individuals with a GG genotype. Meanwhile, through screening of an SSCP technology, excellent individuals with the CC genotype can be screened out through different electrophoretic banding patterns. Thus, the method provided by the invention can be used in production to screen the individuals with the site genotype of CC type as parents for breeding and large-scale breeding of large-sized fries.

Description

technical field [0001] The invention belongs to the technical field of fish genetic selection and breeding, and in particular relates to a SNP site related to the rapid growth of large-sized puffer fish and its application. Background technique [0002] The red-finned puffer (Takifugu rubripes) belongs to the order Telraodontiformes and the genus Fugu. It is mainly distributed in the coast of Japan, and its natural population is small in the coast of my country; and its wild population is in the Therefore, its annual catch is gradually decreasing. The red-finned pufferfish has delicate meat, delicious taste and high protein content. It is known as the "king of fish". Although pufferfish is a poisonous fish, the toxin contained in its liver and ovary is very strong neurotoxic, but its toxin preparation can be used clinically to treat arthritis, rheumatic pain, headache, etc. Therefore, this kind of fish has high comprehensive utilization value. At present, the natural resou...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/156C12Q2600/124C12Q2565/125Y02A40/81
Inventor 王秀利王娟胡子文张峰刘鹰仇雪梅
Owner DALIAN OCEAN UNIV
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