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Molecular marker C2 for identifying sex of penaeus chinensis and application of molecular marker C2

A Chinese prawn, molecular marker technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. Easy to misjudge and identify fast effects

Active Publication Date: 2021-11-26
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, juvenile Penaeus prawns, especially early-stage larvae, are difficult to distinguish between male and female by appearance. At present, there is no method for sex identification of Penaeus chinensis by molecular technology, which greatly hinders the development of parthenogenesis and aquaculture industry

Method used

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  • Molecular marker C2 for identifying sex of penaeus chinensis and application of molecular marker C2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Align the resequencing data of the female Penaeus chinensis pool with the reference genome (based on a male shrimp), and extract the unaligned short sequences for assembly. The assembled result sequence is then compared with the reference genome, and the sequence with more than 60% similarity to the reference genome is deleted. The resequencing data of female mixed ponds and male mixed ponds of Penaeus chinensis were compared with the assembly results, and the sequences with male coverage depth = 0 and female coverage depth > 30 were retained in the assembly results. This result was verified, and the molecular marker C2 was obtained.

[0030] The nucleotide sequence of the molecular marker C2 used for the sex identification of Chinese prawns is shown in SEQ ID No.1. The molecular marker is a female-specific nucleotide sequence with a full length of 1802 bp.

[0031] 2. The present invention utilizes the molecular marker C2 that identifies the female specificity of P...

Embodiment 2

[0038] Samples for testing: 20 "Huanghai No. 1" Chinese prawns with known sex and 20 "Huanghai No. 3" Chinese prawns with known sex, 10 males and 10 females in each group.

[0039] 1. Take the muscle tissue of the test sample, extract the genomic DNA, measure the concentration and dilute to 100 ng / μL.

[0040] 2. Using the genomic DNA obtained in step 1 as a template, perform PCR amplification.

[0041] The sequence of the control primer pair used in PCR amplification is as follows:

[0042] Control-F: 5'-GGGTGGGTGGTATGGAAAGT-3';

[0043] Control-R: 5'-TCCTTAATCCTGCTGCATCCA-3'.

[0044] The sequence of the female-specific primer pair used in PCR amplification is as follows:

[0045] C2-F: 5'-CTTTGGGCTGGCGGTATTTT-3';

[0046] C2-R: 5'-CCAGACTGCAGCTTCCATG-3'.

[0047] The reaction system used for PCR amplification was 15 μL, including 7.5 μL 2×Taq PCR Mix (containing dNTPs, TaqDNA polymerase, MgCl 2 , reaction buffer), PCR control or 0.3 μL of female-specific upstream and ...

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Abstract

The invention discloses a molecular marker C2 for identifying the sex of penaeus chinensis and application of the molecular marker C2. The nucleotide sequence of the molecular marker C2 is as shown in SEQ ID No.1, and the molecular marker C2 is a specific molecular marker for female penaeus chinensis. Meanwhile, a pair of amplification primers with nucleotide sequences as shown in SEQ ID No.2 and SEQ ID No.3 are also obtained by utilizing the molecular marker C2. In the invention, the genome DNA of the penaeus chinensis to be detected is used as a template, an amplification primer pair is used for PCR amplification, then a PCR amplification product is detected by electrophoresis, when a 192 bp specific amplification band appears in the amplification product, the penaeus chinensis is judged to be female, and when no amplification product appears, the penaeus chinensis is judged to be male. The molecular marker C2 and the amplification product thereof which are used for identifying the sex of the penaeus chinensis are simple to operate, accurate in result and not easy to misjudge, and are not limited to the development period of the penaeus chinensis.

Description

technical field [0001] The invention belongs to the field of sex identification of marine shrimps, and in particular relates to a molecular marker C2 for sex identification of Chinese prawns and an application thereof. Background technique [0002] Chinese prawns ( Fenneropenaeus chinensis ) belongs to Arthropoda, Crustacea, Decapoda, Penaeidae, also known as Oriental prawn, commonly known as prawn and prawn. Penaeus chinensis is a large-scale migratory shrimp with wide-temperature, wide-saline and annual warm water. It has obvious migration habits in the Yellow Sea and Bohai Sea. It is divided into winter migration and reproductive migration. The spawning period is between April and June. 300,000 to 1,000,000 grains. [0003] Chinese shrimp is one of the most representative seawater economically farmed animals in China, and it is also one of the most important breeding and export objects in my country from the 1960s to the 1990s. The body length of sexually mature female...

Claims

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Application Information

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IPC IPC(8): C12Q1/6879C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6879C12Q1/6888C12Q1/686C12Q2600/166C12Q2565/125C12Q2545/113Y02A40/81
Inventor 王琼任宪云何玉英吕建建李健
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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