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SNP molecular markers related to traits of grass carp and their application

A technology of molecular markers and grass carp, applied in the direction of recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of typing cost and high cost of measuring traits, so as to save breeding time, high accuracy, The effect of speeding up the breeding process

Active Publication Date: 2021-11-23
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The larger the quantity, the higher the cost of typing and measuring traits

Method used

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  • SNP molecular markers related to traits of grass carp and their application
  • SNP molecular markers related to traits of grass carp and their application
  • SNP molecular markers related to traits of grass carp and their application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Obtaining of SNP markers

[0044]1. The grass carp gene sequence obtained by the sequencing of the present invention is as SEQ ID NO.1~SEQ ID NO.4, in the 159th base of SEQ ID NO.1, the 410th base of SEQ ID NO.1, the 410th base of SEQ ID NO.1 One SNP site was found at the 370th base and the 374th base of SEQ ID NO.1, the 159th base of SEQ ID NO.1, the 410th base of SEQ ID NO.1, and the 410th base of SEQ ID NO. 1 There are alleles C and T at the 370th base and the 374th base of SEQ ID NO.1, forming CC, TC, and TT genotypes.

[0045] The samples used in the analysis of growth traits (body weight) in the present invention are 25-month-old grass carp populations bred in the same batch and reared in the same pond. 298 grass carp were randomly selected for correlation analysis of growth traits (body weight), and 20 individuals with a very large body weight were selected. (average body weight is 2659±126.40g), and 20 extremely small individuals (average body weight ...

Embodiment 2

[0055] Example 2 Screening of Growth Difference Markers

[0056] The 84 differential SNPs markers screened out were preliminarily verified by constructing 20 samples of extremely large individuals and 20 samples of extremely small individuals. Among the 84 selected SNPs loci, no SNP mutations were detected in 40 of them, and SNP mutations were detected in 44. The approach analysis of the difference between the 44 SNPs that detected the mutant type in the extremely large individual group and the extremely small individual group showed that there was a significant difference between SNP24 and SNP36 (P<0.05), and a very significant difference between SNP37 and SNP38 (P<0.01) (see Table 2).

[0057] Table 2 Markers in extreme group typing

[0058]

[0059]

[0060]

[0061] Note: "_" means P<0.05 for this site.

Embodiment 3

[0062] Example 3 Verification of SNP markers associated with body weight in grass carp and body weight

[0063] 1. Sample DNA Extraction

[0064] (1) Take 100ul of blood from the fish to be tested or 3mg of shredded fin ray tissue, add 0.5mL of lysate (10mmol / LTris-HCl; 0.1mol / L EDTA; 0.5% SDS; 30mg / L RNase; 100mg / L Proteinase K, pH 8.0), digested at 55°C for 1 hour with gentle shaking from time to time.

[0065] (2) Add an equal volume of phenol / chloroform / isoamyl alcohol (25:24:1), mix upside down, let stand at room temperature for 5 minutes, centrifuge at 12,000 rpm for 10 minutes, take the supernatant, and then wash with chloroform Extract once, let stand at room temperature for 5 minutes, centrifuge at 12,000 rpm for 10 minutes, and take the supernatant.

[0066] (3) Add 2 times the volume of absolute ethanol, let stand at room temperature for 10 minutes to precipitate DNA, and centrifuge at 12,000 rpm for 10 minutes.

[0067] (4) Wash once with 70% ethanol, centrifuge...

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Abstract

The invention discloses SNP molecular markers related to grass carp traits and their applications. The SNP sites related to the growth status of grass carp are located at the 159th base of grass carp gene SNP24, the 410th base of gene SNP25, and the 370th base of gene SNP26 base, the 374th base of gene SNP27; if the 159th base of gene SNP24, the 410th base of gene SNP25, and the 370th base of gene SNP26 are CC genotype, the body weight is obviously better than that of TT genotype individuals, If the 374th base of gene SNP27 is TT genotype, the body weight is obviously better than that of CC and TC genotype individuals. The invention can be effectively used for the molecular marker assisted breeding of grass carp, select suitable genotype grass carp parents for breeding, can obtain grass carp fry with faster growth, and accelerate the process of grass carp breeding.

Description

technical field [0001] The invention belongs to the field of DNA marker technology and application of molecular biology, in particular to SNP molecular markers related to grass carp traits and applications thereof. Background technique [0002] Grass carp (Ctenopharyngodon idella) belongs to the subfamily Leuciscinae (Leuciscinae) and the genus Ctenopharyngodon. It has the advantages of fast growth, delicious meat, and high nutritional value. From 2007 to 2016, the output of grass carp increased from 3.56 million tons to 5.9 million tons year by year, and the output increased by 65.73% in ten years. It can be seen that the healthy and sound development of the grass carp farming industry not only provides a large amount of high-quality animal protein for the people of our country and even developing countries, but also drives the development and growth of related industries such as feed processing, transportation and sales industries, and solves the employment of a large numb...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/124C12Q1/6869C12Q2531/113C12Q2533/101C12Q2535/122Y02A40/81
Inventor 樊佳佳马冬梅姜鹏白俊杰
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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