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SNP (single-nucleotide polymorphism) marker related to Crassostrea gigas glycogen content character and application thereof

A long oyster and glycogen technology, which is applied in the fields of molecular biology and genetics and breeding, can solve the problems such as rare research on the quality traits of the long oyster, and achieves the effects of avoiding the use of concentrated acid and alkali, and being safe and easy to operate.

Active Publication Date: 2014-11-19
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few studies on the quality traits of long oyster

Method used

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  • SNP (single-nucleotide polymorphism) marker related to Crassostrea gigas glycogen content character and application thereof
  • SNP (single-nucleotide polymorphism) marker related to Crassostrea gigas glycogen content character and application thereof
  • SNP (single-nucleotide polymorphism) marker related to Crassostrea gigas glycogen content character and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Screening of SNP sites related to glycogen content

[0034] a) Collection of samples: A total of 144 wild oysters were collected from wild oyster populations in Jiaonan, Qingdao. They were dissected, and samples were taken by tissue (adductor muscle, gill tissue, etc.), and frozen in liquid nitrogen at -80°C. Save for later.

[0035] b) Detection of glycogen content: the relative content of glycogen in the adductor muscle of each individual was detected with the muscle glycogen and liver glycogen determination kit of Nanjing Jiancheng Bioengineering Institute, and the detection method was carried out according to the operation steps in the manual.

[0036] 1) Sampling: Take the frozen muscle sample, rinse it with normal saline, blot it dry with filter paper, and weigh it (sample weight ≤ 100 mg).

[0037] 2) Hydrolysis: according to the weight of the sample (mg): the volume of lye in the kit (μl) = 1:3, add them together to the test tube, cook in a boiling water bath f...

Embodiment 2

[0070] Application of a SNP marker associated with glycogen content of long oyster

[0071] a) Collection of samples: A total of 96 wild oysters were collected from Shentanggou, Qingdao. They were dissected, gills and adductor muscles were taken, and they were quickly frozen with liquid nitrogen and stored at -80°C for later use.

[0072] b) DNA extraction: the extraction method is the same as c) in Example 1.

[0073] c) SNP site TY202 genotype detection:

[0074] 1) Using the DNA of 96 wild long oysters in Shentanggou as a template, PCR amplification was carried out with specific primers of TY202, and the amplification reaction was carried out in a skirted 96-well PCR reaction plate. Seal with mineral oil, and the reaction conditions are shown in Table 2.

[0075] 2) After the reaction, add 1 μl of internal standard (internal standard as above) and 1 μl of LC-green dye, denature at 95°C for 10 minutes after transient centrifugation, and cool to room temperature.

[0076] ...

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Abstract

The invention relates to an SNP (single-nucleotide polymorphism) marker related to Crassostrea gigas glycogen content character and application thereof. The site of the SNP marker is named TY202 and positioned on the 3rd exon of the glycogen debranching enzyme gene, and the genotype is Y. The TY202 is positioned on the 7022nd base at the 5' end of the glycogen debranching enzyme gene. The 3rd exon is positioned on the 6446-7043 base pair at the 5' end of the glycogen debranching enzyme gene. The glycogen content of the individual with the SNP marker site genotype of T / T or T / C is higher than that of the individual with the genotype of C / C. The method for detecting the individual genotype can be utilized to predict the glycogen content of the individual. When being used in breeding, the SNP marker can be used for selecting the target individual and for molecular marker assisted selective breeding. The SNP marker avoids using concentrated acid or concentrated alkali in the glycogen detection process, thereby being safer and more convenient to operate.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and genetic breeding, in particular to a SNP marker related to the trait of glycogen content in long oyster. Background technique [0002] Long oysters belong to the clambranchia (or bivalves) in the mollusk phylum. The meat is tender and nutritious. Glycogen is an important flavor nutrient in oysters, which can be directly absorbed by the body, thereby reducing the burden on the pancreas. Therefore, it is very effective in the prevention and treatment of diabetes. [0003] Glycogen debranching enzyme is an enzyme in the process of glycogen degradation in oysters. Its main function is to hydrolyze α-1,6 glycosidic bonds, remove branches of glycogen, and make branched glycogen into linear glycogen. Deficiency of human glycogen debranching enzyme can cause glycogen storage syndrome-type 111 disease. At present, the full-length DNA of glycogen debranching enzyme is available. Mutation sc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 佘智彩李莉张国范
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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