Scylla paramamosain biomarker CYP2 gene and application of scylla paramamosain biomarker CYP2 gene in preparation of pathological detection reagent

A technology of biological markers and pseudo-cave blue crabs, which is applied in the fields of plant gene improvement, biochemical equipment and methods, and microbial measurement/inspection, can solve the problems that hinder the healthy development of the blue crab industry, the low output per mu of blue crabs, and the Crab disease outbreaks and other issues

Active Publication Date: 2022-05-27
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

In recent years, due to factors such as blue crab bacterial disease and heavy metal stress in the water environment, the outbreak of blue crab disease has been serious, and the yield of blue crab per mu is not high, which has seriously hindered the healthy development of the blue crab industry.

Method used

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  • Scylla paramamosain biomarker CYP2 gene and application of scylla paramamosain biomarker CYP2 gene in preparation of pathological detection reagent
  • Scylla paramamosain biomarker CYP2 gene and application of scylla paramamosain biomarker CYP2 gene in preparation of pathological detection reagent
  • Scylla paramamosain biomarker CYP2 gene and application of scylla paramamosain biomarker CYP2 gene in preparation of pathological detection reagent

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Embodiment 1

[0021] 1. Materials method

[0022] 1.1 RNA extraction and cDNA synthesis

[0023] Total RNA was extracted from the hepatopancreas of Scylla simulans by sampling Trizol.

[0024] (1) Take 0.1 mg of hepatopancreas tissue for grinding, add 1 ml of Trizol, and let stand on ice for 10 min;

[0025] (2) Centrifuge at 12,000rpm for 5min, take the supernatant, add 200ul chloroform, shake vigorously, and let stand on ice for 5min;

[0026] (3) Centrifuge at 12,000 rpm for 5 min, absorb the upper aqueous phase, add an equal volume of isopropanol, and let stand on ice for 5-10 min;

[0027] (4) Centrifuge at 12,000rpm for 5min, discard the supernatant, add 1ml of 75% alcohol, and let stand on ice for 5-10min;

[0028] (5) Centrifuge at 8,000g for 5min, discard the supernatant, and dry at room temperature;

[0029] (6) Add 50ul DEPC water to dissolve.

[0030] The extracted RNA was measured by spectrophotometer, and the quality of RNA was detected by agarose gel electrophoresis. Qu...

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Abstract

The invention discloses a scylla paramamosain biomarker CYP2 gene and application of the scylla paramamosain biomarker CYP2 gene in preparation of a pathological detection reagent. The scylla paramamosain biomarker CYP2 gene (the nucleotide sequence is shown as SEQ ID NO.1) is cloned from scylla paramamosain, the gene is expressed in hepatopancreas, gill, intestines, blood cells, heart, muscles and stomach tissues, the expression quantity in the heart is the highest, the expression quantity in the hepatopancreas is the lowest, the expression quantity in the intestinal tract is the lowest, and the expression quantity in the stomach tissue is the highest. After the pathogenic bacterium vibrio parahaemolyticus stimulates the scylla paramamosain, the expression quantity of the SpCYP2 gene is remarkably increased, and under the stress of heavy metal cadmium, the expression quantity of the SpCYP2 gene is remarkably increased. The CYP2 gene can be used as a unique biomarker in toxicology application, and has important application value in evaluating the health status of aquatic animals.

Description

Technical field: [0001] The invention belongs to the field of disease prevention and control, in particular to the Scylla simulans biological marker CYP2 gene and its application in the preparation of pathological detection reagents. Background technique: [0002] Scylla crab, commonly known as blue crab, has the characteristics of delicious meat and high nutritional value, and is deeply loved by consumers. Scylla simulans are mainly distributed in the southeast coast of my country and some countries in Southeast Asia. Scylla simulans is one of the most important aquaculture seawater crabs in my country, with a cultured output of more than 168,000 tons. In my country, the cultivation mode of blue crab is mainly based on pond cultivation, and the cultivation mode is relatively extensive. In recent years, due to factors such as bacterial diseases of mud crabs and heavy metal stress in the water environment, the outbreak of diseases of mud crabs has been serious, and the yiel...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12N15/12C12Q1/6888C12Q1/6851C12N15/11
CPCC07K14/43509C12Q1/6888C12Q1/6851C12Q2600/124C12Q2600/158C12Q2531/113Y02A40/81
Inventor 程长洪郭志勋马红玲刘广鑫冯娟邓益琴
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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