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Primer and method for quickly detecting gender difference of juvenile fishes of fugu rubripes

A technology for the sex of redfin puffer and juvenile fish, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc.

Active Publication Date: 2014-12-03
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

So far, there have been no reports on the allele-specific PCR method and the rapid detection molecular biology kit for the single base mutation of the sex difference in redfin puffer juveniles at home and abroad

Method used

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  • Primer and method for quickly detecting gender difference of juvenile fishes of fugu rubripes

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Embodiment 1

[0031] The method for rapidly detecting the sex difference of puffer puffer juveniles described in the present invention comprises the following steps:

[0032] 1. Sample collection

[0033] In this example, 20 puffer juveniles of different sexes were used as the research objects, and all 20 individuals were randomly selected from a group of puffer juveniles cultured by an aquatic company. The gonad tissues of each individual were fixed in Bouin’s solution for 24 hours, rinsed with 50% ethanol and stored in 70% ethanol; the fin ray tissues were cut and stored at -20°C.

[0034] 2. Gonad tissue section and sex differentiation identification of redfin puffer juvenile

[0035] The fixed gonad tissue samples of juvenile redfin puffer were sliced ​​by conventional methods, with a thickness of 4-7 μm, and stained with H.E. Observe and take pictures with OLYMPUS DP72 microscope to measure the size of germ cells.

[0036] The oocyte is oval in shape, significantly larger than the o...

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Abstract

The invention discloses a primer and a method for quickly detecting gender difference of juvenile fishes of fugu rubripes. A pair of outer PCR (Polymerase Chain Reaction) primers which respectively amplify gender difference gene sequences and an inner PCR primer which contains mispaired basic groups aiming at mutation sites are designed aiming at G-C mononucleotide mutation sites existing on gender differential genes of male and female individuals of fugu rubripes. By means of the primer, different amplification results can be obtained by different gender DNA (Deoxyribonucleic Acid) samples of fugu rubripes, so that the gender difference of fugu rubripes can be judged. The primer disclosed by the invention is low in cost, simple to operate, quick and accurate and suitable for popularization and application, can quickly identify gender of juvenile fishes of fugu rubripes on a basis of extracting genome DNA of fugu rubripes one year old, and has important application value to accelerate the unisexuality breeding process of fugu rubripes and PCR detection method research in the field of molecular biology.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a primer and a method for rapidly detecting the sex difference of puffer puffer juveniles. Background technique [0002] Redfin Pufferfish ( Takifugu rubripes ) is the most valuable seawater fish in the order Fuguformes. Although it is a poisonous fish, the folk custom of eating puffers in Japan, South Korea and my country has a long history. The red-finned fugu is even regarded as the "king of fish" and "the first of a hundred fishes". Its sashimi is unique and expensive, and it has always been an important representative of Japanese food culture. Pufferfish not only has extremely high edible value, but Tetrodotoxin (TTX) and its preparations contained in its ovary, liver, blood and other tissues are high-grade drugs for relieving muscle spasms and analgesia, and are sold at international prices Up to RMB 5~7×10 4 Yuan / g, its outer skin can also be used for tanning, and its breedi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 刘新富刘滨胡鹏孟振陈松林
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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