Cynoglossus semilaevis sex chromosome-linked DNA segments and application thereof

A semi-smooth tongue sole, chromosome technology, applied in DNA/RNA fragments, recombinant DNA technology, microbial determination/inspection, etc., can solve the problems of high requirements for experimental equipment and reagents, increased costs, long operation time, etc., to save detection time and Cost, increased female ratio, effect of shortened time

Active Publication Date: 2016-02-17
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

Although co-dominant microsatellite sex-specific markers can accurately identify ZZ males and ZW females, because there is only a 12bp difference between the two DNA fragment bands, they cannot be resolved by agarose gel electrophoresis. However, the operation of PAGE gel e...

Method used

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  • Cynoglossus semilaevis sex chromosome-linked DNA segments and application thereof
  • Cynoglossus semilaevis sex chromosome-linked DNA segments and application thereof
  • Cynoglossus semilaevis sex chromosome-linked DNA segments and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Example 1 Screening of Sex Chromosome-linked DNA Fragments of Semi-smooth Tongue Sole

[0021] The sex chromosome-linked DNA fragment sequence used in the present invention is derived from the genome sequencing results of the half-smooth tongue sole completed by the Aquatic Genome and Cell Engineering Laboratory of the Yellow Sea Fisheries Research Institute of the Chinese Academy of Fishery Sciences. Bioinformatics analysis of the half-smooth tongue sole genome sequence screened out two Z, W chromosome homologous differential DNA fragments. Multiple pairs of primers were designed, and a pair of primers (SEQZW:1, SEQZW:2) with stable results were screened out.

[0022] Select the DNA of female and male half smooth tongue sole with known genetic sex, and use primers SEQZW: 1 and SEQZW: 2 for PCR amplification. The reaction conditions and procedures are as follows: PCR reaction system includes 35.5 μL ddH2O; 5.0 μL 10× Buffer; 4.0 μL dNTP (2.5 mmol / L); 1.0 μL SEQZW:1 (1...

Embodiment 2

[0023] Example 2: Application of Sex Chromosomal Linked DNA Fragment of Semi-smooth Tongue Sole

[0024] 1. Genetic sex identification of tongue sole under laboratory conditions

[0025] The sex chromosome-linked DNA fragment primers SEQZW:1 and SEQZW:2 of tongue sole were used to detect the genetic sex of tongue sole by PCR method. The PCR reaction system is 15 μL, including 1.5 μL of 10×buffer, 0.8 μL of dNTP, 0.3 μL of upstream and downstream primers, 1 μL of template DNA (50 ng / μL), 11.0 μL of ddH2O, and 0.1 μL of rTaq enzyme. PCR amplification program: 10min at 95°C; 35 cycles of 95°C for 30s, 60°C for 30s, and 72°C for 30s; 10min at 72°C, and store at 4°C. Add 10×LoadingBuffer2.0μL, 1.5% agarose gel electrophoresis, 150V, 20 minutes, gel imaging can distinguish genetic gender. ZW females contain two DNA bands of 366bp and 253bp, and ZZ males only have one DNA band of 366bp, such as figure 2 shown.

[0026] 2. Genetic sex identification and pseudo-male screening of h...

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Abstract

The invention provides a cynoglossus semilaevis sex chromosome-linked DNA segment and application thereof. A sequence of a DNA segment on a Z chromosome is SEQ ID NO:1; a sequence of a DNA segment on a W chromosome is SEQ ID NO:2. The DNA segment is used for designing a molecular marker for identifying the genetic sex of cynoglossus semilaevis. Two Z-W chromosome homologous and different DNA segments are screened from cynoglossus semilaevis whole genome information, a primer is designed for identifying the genetic sex of the cynoglossus semilaevis, and the genetic sex of the cynoglossus semilaevis can be distinguished quickly, accurately and effectively. By adopting a method, specific purpose stripes can be amplified in male and female individuals and can be distinguished by agarose electrophoresis, the time for accurately identifying the genetic sex of the cynoglossus semilaevis is shortened, the method is suitable for identifying the genetic sex of the cynoglossus semilaevis in a simple environment of a farm, and detection time and cost are reduced.

Description

technical field [0001] The invention belongs to the technical field of fish genetic sex identification and sex control in aquatic biology technology, and in particular relates to a sex chromosome linked DNA fragment of tongue sole and its application in rapid identification of genetic sex of tongue sole. Background technique [0002] Cynoglossus semilaevis (Cynoglossus semilaevis), commonly known as "sole rice" and "sole order", belongs to the order Pleuriformes, Cynoglossus semilaevis, and the genus Cynoglossus semilaevis. The Bohai Sea is the most, followed by the Yellow Sea (Liu Xuezhou et al. 2005; Meng Qingwen et al. 1995). Semi-smooth tongue sole has tender and delicious meat, high nutritional value, and is increasingly popular among people. It has low trophic level, mild food habits, small activity range, and is convenient for factory farming and management. It has broad breeding prospects. Excellent breeding objects (Liu Xuezhou et al. 2005). [0003] tongue sole h...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124
Inventor 陈松林董忠典张宁刘洋邵长伟徐文腾
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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