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Method for identifying genetic sex of rana dybowskii by TRAP molecular marker technology

A molecular marker and gender technology, applied in recombinant DNA technology, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problem of inability to identify the genetic sex of Northeast wood frog, unable to successfully obtain sexual reversal individuals, and no Northeast wood frog yet. Sex-specific DNA molecular markers and other issues

Pending Publication Date: 2020-04-07
NORTHEAST AGRICULTURAL UNIVERSITY
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Problems solved by technology

The physiological sex of the Northeast Rana can be identified through external morphological observation, but because the heteromorphic sex chromosomes were not observed in the karyotype analysis of the Northeast Rana, the genetic sex of the Northeast Rana cannot be identified, so that the sex-reversed individuals cannot be successfully obtained
However, there have been no reports of sex-specific DNA molecular markers in Rana sinensis

Method used

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  • Method for identifying genetic sex of rana dybowskii by TRAP molecular marker technology
  • Method for identifying genetic sex of rana dybowskii by TRAP molecular marker technology

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Embodiment Construction

[0019] The present invention will be further described below in conjunction with embodiment

[0020] Depend on figure 1 It can be seen that the agarose electrophoresis test results of part of the DNA extracted in this experiment show that the DNA fragments are intact, there is no large amount of protein in the sample wells, and the DNA is not significantly degraded, indicating that the amount of DNA is relatively high, which can meet the needs of further analysis.

[0021] Depend on figure 2 It can be seen that lane 17 is a marker of 2000bp; lanes 1-16 are physiological male Rana, and lanes 18-33 are physiological female Rana. In addition to bands that appear in both female and male individuals, lanes 1, 2, 3, 5, 7, 8, 9, 10, 11, 12, 13, 14, 15, and 16 have a sex-specific band of 261 bp strip, ie figure 2 The band shown in the box, the individual with this band is a genetic male individual. This band does not appear in all physiological female individuals, that is, lane...

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Abstract

The invention, which relates to the field of molecular markers, discloses a molecular marker for identifying genetic sex of rana dybowskii and application thereof, thereby solving a problem that the genetic sex of the rana dybowskii cannot be identified at present. The genome of the toe tissue of the rana dybowskii is extracted and PCR amplification reaction is performed by using the extracted genome DNA as a template and a primer pair containing sequences shown in SEQ ID NO: 1 and SEQ ID NO: 2; electrophoresis detection is performed on a PCR amplification product by using 14% of polyacrylamide gel; and the amplification result shows that the individual with a band corresponding to a nucleotide sequence (261 bp) shown in SEQ ID NO: 3 is a genetic male northeast rana dybowskii and the individual without a band corresponding to the nucleotide sequence shown in SEQ ID NO: 3 in the amplification result is a genetic female northeast rana dybowskii. Therefore, pseudo-male individuals, namelyindividuals which are physiologically male but inherited as female, can be identified by the method. Pseudo-male individuals and female individuals are used for mating, and the physiological female proportion of the generated progeny reaches up to over 90%. The sex of the rana dybowskii breeding group can be controlled, and the economic benefit of rana dybowskii breeding is improved.

Description

technical field [0001] The invention relates to the field of molecular markers, in particular to a molecular marker for identifying the genetic sex of the Northeast Rana and its application. Background technique [0002] Northeast Rana ( Rana Dybowskii ) belongs to the amphibian class (Amphibian) Anura (Anura) frog family (Ranidae) frog genus ( Rana ), mainly distributed in the three northeastern provinces of my country and parts of Inner Mongolia, and is one of the important economic animals in Northeast my country. The economic value of Northeast Rana is mainly reflected in the oviduct of the female frog. "Chinese Pharmacopoeia" records that the oviduct of Northeast Rana Rana can be dried to obtain Rana oil. Rana oil is sweet, salty and flat, and returns to the lung and kidney meridian. It has the effects of nourishing kidney and essence, nourishing yin and moistening lung. Weakness, fatigue, palpitations, insomnia, night sweats, tuberculosis and coughing up blood. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6879C12Q1/6888C12N15/11
CPCC12Q1/6879C12Q1/6888
Inventor 许愿狄生伟宁方勇白秀娟杜智恒刘珈羽苏杭徐逸男吕晶
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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