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Molecular marker for identifying genetic sex of rana dybowskii and application method thereof

A molecular marker and gender technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of inability to identify the genetic sex of Northeast wood frogs, and no known sex-specific DNA molecules of Northeast wood frogs. Problems with labeling, inability to successfully acquire individuals with sexual reversal, etc.

Pending Publication Date: 2020-08-28
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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  • Molecular marker for identifying genetic sex of rana dybowskii and application method thereof
  • Molecular marker for identifying genetic sex of rana dybowskii and application method thereof

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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. Lanes 1, 2, 3, 5, 7, 8, 9, 10, 11, 12, 13, 14, 15, and 16 have a 222bp sex-specific band in addition to bands that appear in both female and male individuals belt, 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, lanes 18-3...

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Abstract

The invention provides a molecular marker for identifying genetic sex of rana dybowskii. The method is mainly characterized by comprising the following steps: extracting a genome of a toe tissue of rana dybowskii, and carrying out PCR (Polymerase Chain Reaction) amplification reaction by taking an extracted genome DNA (Deoxyribonucleic Acid) as a template and a primer pair containing sequences asshown in SEQ ID NO:1 and SEQ ID NO:2; and carrying out electrophoresis detection on a PCR amplification product through 14% polyacrylamide gel, wherein the individuals with bands corresponding to a nucleotide sequence (222bp) as shown in SEQ ID NO:3 are genetic male rana dybowskii, and the individuals without the bands corresponding to the nucleotide sequence as shown in the SEQ ID NO:3 are genetic female rana dybowskii. The method has the advantages that the genetic male specific DNA molecular marker of the rana dybowskii is obtained, and the genetic sex of the rana dybowskii can be quickly,simply, conveniently and accurately identified. Pseudo-male individuals, namely the individuals which are physiologically male but inherited as females, can be obtained. The pseudo-male individuals are used for mating with female individuals, and the proportion of physiologically female progenies is as high as 90% or above, so that economic benefits of rana dybowskii breeding can be remarkably improved.

Description

technical field [0001] The invention relates to a method for identifying the genetic sex of the Northeast Rana by molecular markers. Background technique [0002] Northeast Rana ( Rana Dybowskii ) belongs to the amphibian class (Amphibian) Anura (Anura) frog family (Ranidae) frog genus ( Rana ). The dried oviduct of the female Northeast Rana, the Chinese Rana described in the "Dictionary of Traditional Chinese Medicine", is called Hatoad Oil or Rana Oil, which is a long-established nourishing traditional Chinese medicine. Studies have shown that regular consumption of wood frog oil can supplement 18 essential amino acids needed by the human body, enhance physical strength and memory, and enhance immune function against various diseases. [0003] Driven by huge economic interests, people have long carried out "extinct" capture of wild Northeast Rana. As the wild resources of Rana are caught less and less, the individuals are smaller and smaller, and the market demand is ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2565/125
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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