Eriocheir sinensis HSP90 gene clone and application thereof

A Chinese mitten crab, gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve problems such as crab disease, heavy metal ions, and harmful pathogenic bacteria exceeding the standard.

Inactive Publication Date: 2017-01-04
LIAONING OCEAN & FISHERIES SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, with the acceleration of my country's industrialization process, the problems of excessive heavy metal ions and harmful pathogenic bacteria in the breeding water of Chinese mitten crab have become more and more prominent, leading to more and more serious problems such as diseases of the crab, which have become restrictions on the Chinese mitten crab aquaculture industry. The main issues of sustainable and healthy development

Method used

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  • Eriocheir sinensis HSP90 gene clone and application thereof
  • Eriocheir sinensis HSP90 gene clone and application thereof
  • Eriocheir sinensis HSP90 gene clone and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0035] The full-length cDNA sequence amplification of embodiment 1 Chinese mitten crab HSP90 gene

[0036] In the present invention, the Chinese mitten crab used in the examples is collected from the mouth of Shuangtaizi River in Panjin, Liaoning Province.

[0037]1. Extraction of total RNA from Eriocheir sinensis

[0038] Taken from the wild Chinese mitten crab (about 70 grams in weight) in Panjin, temporarily kept in the breeding room, the liver and pancreas of the Chinese mitten crab were dissected live with dissecting scissors, ground in liquid nitrogen, and extracted using the TIANGEN animal tissue total RNA extraction kit. Extraction of total RNA, nucleic acid protein analyzer (NanoPhotometer) and 1.0% agarose gel electrophoresis to detect its quality and integrity.

[0039] 2. Synthesis of cDNA

[0040] The total RNA extracted from the hepatopancreas tissue of Eriocheir sinensis was used as a template, and the Primescript RT ReagentKit (TaKaRa Company) kit was used fo...

Embodiment 2

[0055] Example 2. Stress expression test and in vitro recombinant expression of Eriocheir sinensis HSP90 gene

[0056] (1) Expression of HSP90 gene stimulated by different tissues and bacteria

[0057] 1. Experimental materials and pathogenic bacteria

[0058] The wild Chinese mitten crab (about 70 grams in weight) was taken from Panjin, and it was temporarily raised in the breeding tank in the breeding room for a week, the water temperature was 19°C, the pH was 8.0-8.2, the water was changed every day, and fresh clams and Philippine clams were fed.

[0059] Staphylococcus aureus (Staphylococcus aureus) and Vibrio anguillarum (Vibrio anguillarum) used in the challenge test were preserved by our laboratory, cultured overnight in liquid LB at 37°C before use, centrifuged at 3000×g for 20min, removed the supernatant, and washed with TBS (10mM Tris -HCL and 150mM NaCL; Ph 7.5) washed once, and the bacteria were diluted to 10 per ml with TBS 8 CFU / mL for use.

[0060] 2. Immunos...

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Abstract

The invention discloses an eriocheir sinensis HSP90 gene clone and application thereof. The gene has a base sequence as shown in SEQ ID NO.1. The protein encoded by the gene has an amino acid sequence as shown in SEQ ID NO.2. The HSP90 gene provided by the invention is firstly obtained from eriocheir sinensis hepatopancreas by amplification, can be used for in vitro recombination expression and gene transfer of the HSP90 gene, and lays a foundation for developing medicine products, selected genetic breeding and improving, and enhancing stress resistance to high temperature, low temperature, heavy metal ion exceeding, pathogenic bacterial infection and the like.

Description

technical field [0001] The invention relates to the amplification and prokaryotic expression of a heat shock protein 90 (HSP90) gene cDNA full-length sequence of Chinese mitten crab. Background technique [0002] Chinese mitten crab (Eriocheir sinensis), also known as river crab, hairy crab, and hairy crab, is widely distributed in the Liaohe River, Yellow River, Yangtze River, Oujiang River and Minjiang River in my country. The most productive freshwater crab among crabs. It grows fast, the meat is tender and delicious, and it is an important economic aquaculture species in my country. The cultivation of this crab has become the pillar industry of aquaculture in my country. However, with the acceleration of my country's industrialization process, the problems of excessive heavy metal ions and harmful pathogenic bacteria in the breeding water of Chinese mitten crab have become more and more prominent, leading to more and more serious problems such as diseases of the crab, wh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/435C12N15/70C12N1/21C12N15/10A61K39/39A61P37/04C12R1/19
CPCC07K14/43509A61K39/39C12N15/70C12N2800/101C12Q1/6806C12Q2531/113
Inventor 高祥刚高磊鲍相渤赫崇波刘卫东苏浩李云峰
Owner LIAONING OCEAN & FISHERIES SCI RES INST
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