A kind of oyster cell apoptosis gene caspase 3 gene and its application in the preparation of pathological detection diagnostic reagents

A technology for detecting reagents and oysters, which can be used in applications, genetic engineering, plant genetic improvement, etc., and can solve problems such as output fluctuations, high energy consumption, and large breeding areas

Active Publication Date: 2021-03-19
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because its breeding method is mainly traditional farming, there are limitations such as large bree...

Method used

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  • A kind of oyster cell apoptosis gene caspase 3 gene and its application in the preparation of pathological detection diagnostic reagents
  • A kind of oyster cell apoptosis gene caspase 3 gene and its application in the preparation of pathological detection diagnostic reagents
  • A kind of oyster cell apoptosis gene caspase 3 gene and its application in the preparation of pathological detection diagnostic reagents

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Embodiment 1

[0031] 1. RNA extraction and reverse transcription

[0032] a). Hong Kong oyster cells or tissues were added with Trizol, ground with a tissue grinder, and left at room temperature for 5 minutes to fully lyse.

[0033] b). Centrifuge at 12,000rpm for 5min and discard the precipitate.

[0034] c). Add chloroform to 200ul chloroform / ml Trizol, vortex and mix well, and place at room temperature for 15min.

[0035] d). Centrifuge at 12,000 g for 15 min at 4°C.

[0036] e). Aspirate the upper aqueous phase into another centrifuge tube.

[0037] f). According to 0.5ml isopropanol / ml Trizol, add isopropanol and mix well, and place at room temperature for 5-10min.

[0038] g). Centrifuge at 12,000g at 4°C for 10 minutes, discard the supernatant, and sink the RNA to the bottom of the tube.

[0039] h). Add 75% ethanol to 1ml 75% ethanol / ml Trizol, shake the centrifuge tube gently, and suspend the precipitate.

[0040] i). Centrifuge at 8,000g at 4°C for 5 minutes, and discard the ...

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Abstract

The invention discloses an oyster cell apoptosis gene caspase 3 gene and an application thereof in preparation of a pathological detection and diagnosis reagent. A Hong Kong crassostrea gigas cell apoptosis marker gene-caspase 3 gene is found out, has the nucleotide sequence shown in SEQ ID NO.1, is significantly up-regulated after being infected by vibrio alginolyticus and staphylococcus haemolyticus, and can be used as a standard for detection of oyster cell apoptosis conditions; a positive and accurate warning signal is provided for prevention of oyster disease, and the expansion of disasters is prevented. With the application of the oyster cell apoptosis gene caspase 3 gene, the germplasm innovation ability of cultured shellfishes can be further enhanced and the efficient and sustainable development of the industry is supported.

Description

Technical field: [0001] The present invention relates to the molecular mechanism of shellfish immunity and cell apoptosis, and more specifically relates to a nucleotide sequence of caspase 3 gene screened from Hong Kong giant oyster (Crassostrea hongkongensis) blood cell differential hybridization library stimulated by Vibrio, Amino acid sequence, its preparation method and the use of the gene. Background technique: [0002] my country's annual oyster production accounts for more than 80% of the world's total production, and it is the largest oyster farming country in the world. Hong Kong oysters are widely distributed in coastal waters from Guangxi to Fujian, and are the most important species of oysters cultured along the coast of South China. Because its breeding method is mainly traditional farming, there are limitations such as wide breeding area and high energy consumption, and its output also fluctuates due to changes in various conditions. As the marine environment...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12C12Q1/6883C12N15/11
CPCC07K14/43504C12Q1/6883C12Q2600/158C12Q2600/166Y02A50/30
Inventor 向志明秦艳平马海涛李军张扬肖述张跃环喻子牛
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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