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A kind of semi-smooth tongue sole sex-specific scar marker and application method

A semi-smooth tongue sole and sex technology is applied to the semi-smooth tongue sole sex-specific SCAR marker and its application in the rapid identification of hereditary sex, which can solve the problem of high requirements for experimental equipment and reagents, increased cost of genetic sex identification, and inability to distinguish ZW females and WW superfemales. individual problems, to achieve the effect of improving work efficiency and operability, improving breeding output and economic benefits, and promoting sustainable and healthy development

Active Publication Date: 2016-04-20
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in practical applications, both of these tags have their own shortcomings
Due to its dominant genetic characteristics, AFLP markers cannot distinguish ZW females from WW superfemale individuals, and in actual identification, there may be misjudgment of ZZ males and ZW females due to false negatives
Although codominant microsatellite sex-specific markers can accurately identify ZZ males, ZW females, and WW superfemale individuals, they cannot be distinguished by agarose gel electrophoresis because of the small difference between the two DNA fragment bands. It can be identified by the method of polyacrylamide gel (PAGE gel) electrophoresis with higher resolution, but the operation of PAGE gel electrophoresis is difficult, the operation time is long, the requirements for experimental equipment and reagents are high, and the genetic sex identification is increased. Cost, difficulty and time consuming, it is difficult to conduct on-site testing in tongue sole farms

Method used

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  • A kind of semi-smooth tongue sole sex-specific scar marker and application method
  • A kind of semi-smooth tongue sole sex-specific scar marker and application method
  • A kind of semi-smooth tongue sole sex-specific scar marker and application method

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Embodiment Construction

[0020] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0021] 1. Screening of sex-specific microsatellite markers in tongue sole

[0022] The microsatellite marker sequence used in the present invention is derived from the whole genome sequencing of 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, and then 159 sex-specific microsatellites were screened from 3000 microsatellite sites. Satellite markers. It contains 122 Type B loci (one band in females, none in males) and 37 Type A loci (two bands in females and one band in males).

[0023] Four DNA samples of female and male half smooth tongue soles with known sex were taken, and 37 Type A loci were genotyped by polyacrylamide gel electrophoresis to distinguish the difference between male and female bands. The...

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Abstract

The invention first provides a Cynoglossus semilaevis gender specific microsatellite marker, and comprises a microsatellite marker located in the Z chromosome, and the nucleotide sequence is SEQ ID NO:1. The invention also provides primers designed based on the above microsatellite marker, and the sequences of the upstream primer and the downstream primer are SEQ ID NO:2 and SEQ ID NO:3 respectively. The invention screens a Cynoglossus semilaevis gender specific microsatellite marker, and the Cynoglossus semilaevis gender specific microsatellite marker is subjected to SCAR conversion. Primers marked by SCAR are designed for Cynoglossus semilaevis heredity gender identification. Female, male and superfemale individuals of Cynoglossus semilaevis can be distinguished rapidly, accurately and effectively. Because the marker has characteristics that objective straps can be amplified in females and males and the objective straps can be distinguished with agarose electrophoresis, the time for accurate identification of Cynoglossus semilaevis heredity genders is shortened obviously, the on-site batch identification of Cynoglossus semilaevis heredity genders in a culturing farm of Cynoglossus semilaevis can be carried out, the detection cost is saved, and the work efficiency and accuracy of on-site detection of Cynoglossus semilaevis heredity genders in a culturing farm are raised.

Description

technical field [0001] The invention belongs to the technical field of genetic sex identification and sex control of fish in aquatic biology technology, and in particular relates to a sex-specific SCAR marker of half-smooth tongue sole and its application in rapid identification of genetic sex. Background technique [0002] Cynoglossus semilaevis (Cynoglossus semilaevis) is a large bottom-dwelling fish endemic to my country's coastal waters, and it is distributed in my country's coastal areas mainly in the Yellow Sea and Bohai Sea. Semi-smooth tongue sole has tender meat, delicious taste, and rich nutrition. It has broad breeding prospects and is an important and valuable marine fish. [0003] Production practice shows that there is a huge difference in growth speed between female and male fish of semi-smooth tongue sole, and the growth speed of females is 2-4 times that of males. The characteristics of slow growth and small size of tongue sole male fish reduce the producti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 陈松林刘洋高峰涛孟亮邵长伟宋文涛
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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