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Molecular marker for identifying genetic sex of hucho taimen and identifying method

A technology of molecular markers and salmon, which is applied in the field of genetic sex identification of fish, can solve the problems of crisis, poor wild resources, and the inability to identify sex of fish, and achieve the effect of convenient operation and small sample size

Active Publication Date: 2019-08-30
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Jerome salmon is a rare and precious fish, so it is often necessary to know the sex ratio of wild resources during field investigations, but Jerome salmon below the age of 3 cannot be sexed, and usually fish over the age of 3 need to kill the fish and take the gonads. Only slices can be used to identify males and females, which will endanger the already poor wild resources, so there is an urgent need for a method that can be used to identify males and females without killing fish

Method used

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  • Molecular marker for identifying genetic sex of hucho taimen and identifying method

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Experimental program
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specific Embodiment approach 1

[0016] Specific implementation mode one: the method for identifying the genetic sex of salmon in this implementation mode:

[0017] 1. Extract the genomic DNA of the sample to be identified;

[0018] 2. Perform PCR amplification using genomic DNA as a template in step 1. In the PCR amplification system, the upstream primer of the molecular marker is 5'-ATCAATCATGGCTGGAGGTG-3', and the downstream primer of the molecular marker is 5'-CACCCACACACGTACCAGTC-3';

[0019] 3. Gel electrophoresis of the PCR product, which shows that one band appears to be the female Jerome salmon, and that which shows two bands is the Jerra salmon male.

specific Embodiment approach 2

[0020] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the PCR amplification reaction system in step 2 is 20 μl: 2×Taq PCR mix 10 μl, the genomic DNA of the sample to be identified is 2 μl, and the concentration of molecular marker primers is 10 μM, 1 μl each of the upstream and downstream primers of the molecular marker, and the rest was supplemented with enzyme-free ultrapure water; the PCR reaction was: pre-denaturation at 95°C for 3 minutes, followed by 30 cycles of denaturation at 95°C for 30 sec, annealing at 60°C for 30 sec, and extension at 72°C for 30 sec; Extend at 72°C for 5 min. Others are the same as the first embodiment.

[0021] Under the PCR amplification reaction conditions of this embodiment, the target sequence is completely denatured without affecting the activity of the Taq enzyme, and the primers are completely extended to achieve an effective amplification amount.

specific Embodiment approach 3

[0022] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is:

[0023] Step 1: Take the 0.4cm jelly salmon to be identified 2 The fin ray samples were lysed with 100μl lysate, and then digested with a PCR instrument. The digestion process was 55°C for 2h and 98°C for 10min; after that, mix well, and then centrifuge at 1000-2000rpm for 1-2min, and take the supernatant to get Sample genomic DNA. Others are the same as the first or second embodiment.

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Abstract

The invention relates to a molecular marker for identifying genetic sex of fishes and an identifying method, in particular to a molecular marker for identifying genetic sex of hucho taimen and an identifying method. A nucleotide sequence of specific stripes of male hucho taimen in the molecular marker for identifying genetic sex of hucho taimen is shown in SEQ ID NO:1, and the nucleotide sequenceof common stripes of male hucho taimen is shown in SEQ ID NO:2. The indentifying method comprises the steps as follows: 1, extraction of genomic DNA; 2, PCR amplification; 3, gel electrophoresis: hucho taimen showing one stripe is female while hucho taimen showing two stripes is male. By use of the method, the sex of juvenile hucho taimen can be rapidly and efficiently identified, only a pair of primers is needed for amplification, the sex of the hucho taimen can be identified without sequencing according to the number of stripes of an agarose gel electrophoresis amplified product.

Description

technical field [0001] The invention relates to a molecular marker for identifying genetic sex of fish and an identification method. Background technique [0002] Hucho taimen (pallas) belongs to the order Salmoniformes, the family Salmonidae, and the genus Hucho. It is a rare and valuable cold-water fish in my country. Maturity is late, females reach sexual maturity in 5-6 years, males reach sexual maturity in 4-5 years, without secondary sexual characteristics. Sexually mature salmon have marriage color only during the breeding season, and the tail of the fish body is red, and the marriage color of some parent fish is not obvious, so it is difficult to identify gender by marriage color. Although there is not much difference in growth rate between male and female salmon, the roe of salmon is rich in nutrients and can be used to make caviar, and the commodity price is far higher than that of male fish; and during artificial insemination, the semen of a male fish can be compa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6879C12N15/11
CPCC12Q1/6888C12Q1/6879Y02A40/81
Inventor 匡友谊唐国盘董乐杨笑星佟广香尹家胜张庆渔
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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