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SNP locus related to rapid growth of takifugu obscurus and application thereof

A fugu obscura, site technology, applied in the field of fish genetic selection and breeding, can solve the problem of lack of auxiliary molecular marker technology

Active Publication Date: 2019-07-12
DALIAN OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of auxiliary molecular marker technology that can be applied in the field of puffer obscurus breeding

Method used

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  • SNP locus related to rapid growth of takifugu obscurus and application thereof
  • SNP locus related to rapid growth of takifugu obscurus and application thereof
  • SNP locus related to rapid growth of takifugu obscurus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Cloning of Fugu obscurus MSTN gene nucleotide sequence

[0018] Myostatin gene (Myostatin, MSTN), also known as growth and differentiation factor 8 (growth and differentiation factor 8), belongs to the superfamily of transforming growth factor beta (transforming growth factor beta), and is a negative regulator of muscle growth and development.

[0019] The step of cloning MSTN gene of the present invention is as follows:

[0020] a) Extraction of Fugu obscurus genome. The genomic DNA in the fin rays of Puffer puffer obscura was extracted by conventional phenol imitation method. Take 120mg of pufferfish obscura tail fin tissue in a 1.5ml centrifuge tube, and cut it into pieces with ophthalmic scissors; add 700 μL of DNA extraction solution to the centrifuge tube that has cut the tail fin; add 5 μL of proteinase K (20 mg / ml), gently Gently mix and digest in a 55°C water bath for 2 hours; cool the solution to room temperature, add an equal volume of phenol, ext...

Embodiment 2

[0026] Example 2: Screening to obtain SNP sites

[0027] Determining the location of the SNP associated with the puffer puffer MSTN gene and growth traits comprises the following steps:

[0028] a) Acquisition of Fugu obscurus samples: The population used was a family of Fugu obscurus hatched by the farm of Dalian Tianzheng Industrial Co., Ltd., and 109 Fugu obscurus individuals were randomly selected from this family.

[0029] b) Data collection and genomic DNA extraction: The body weight, body length and body length phenotype values ​​of 109 individuals were measured and recorded, and part of the caudal fin rays were cut for genomic DNA extraction.

[0030] c) PCR reaction and sequencing. The reaction system is 25 μl: 10×buffer 2.5 μl, dNTP 2 μl, SNP-f primer (SEQ ID NO.2) 1 μl, SNP-r primer (SEQ ID NO.3) 1 μl, genomic DNA 1 μl, EX Taq enzyme 0.25 μl, double Make up to 25 μl with distilled water. The PCR reaction program was: 98°C for 30s; 30 cycles of 98°C for 10s, 65°C ...

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Abstract

The invention provides an SNP locus related to rapid growth of takifugu obscurus and application. The SNP locus is located at position 724 of an MSTN gene with a nucleotide sequence shown as SEQ ID NO:1, and the base of the SNP locus is C or T. The SNP locus is used for breeding takifugu obscurus individuals with rapid growth potential. According to the method, by analyzing the correlation betweenthe locus genotype frequency and the growth traits of the takifugu obscurus, the SNP locus related to the growth traits is found at 724 base position of the MSTN gene of the takifugu obscurus, and the body weight, the body length and the body full length of individuals with TT homozygotic genotypes are obviously greater than phenotypic values of the growth traits of individuals with CC and CT genotypes (p smaller than 0.05). Thus, individuals with TT genotype at the locus can be preferentially selected as parents or for large-scale breeding in production.

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 puffer puffer obscurus and its application. Background technique [0002] Fugu obscurus (Takifugu obscurus) belongs to Fuguformes, Fuguidae, Fugu genus, mainly distributed in the middle and lower reaches of the Yangtze River in my country, Bohai Sea, Yellow Sea and other places. Pufferfish obscurus has delicious meat and rich nutritional value. There is a saying in ancient folks that "eat pufferfish desperately". Since October 2016, my country has conditionally released the production and breeding of red-finned puffer and puffer obscurus. Fugu obscurus grows very slowly, significantly smaller than puffer red fin. The average body length of the 2-year-old cultured red-finned puffer is about 30.4cm, and the average weight is about 1062g, while the average body length of the cultured puffer obscurus ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156Y02A40/81
Inventor 仇雪梅窦冬雨王秀利
Owner DALIAN OCEAN UNIV
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