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Novel shellfish pattern recognition receptor LRPP (Leucine-rich Repeat Protein) specifically combined with LPS (Lipopolysaccharide)

A technology for pattern recognition of receptors and shellfish, applied in the direction of receptors/cell surface antigens/cell surface determinants, specific peptides, animal/human peptides, etc., can solve problems such as shortage of good breeds, death, disease outbreaks, etc.

Active Publication Date: 2014-06-18
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the lack of good species leads to low immunity of oysters, disease outbreaks, and even mass mortality, which is the main bottleneck restricting the development of the industry

Method used

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  • Novel shellfish pattern recognition receptor LRPP (Leucine-rich Repeat Protein) specifically combined with LPS (Lipopolysaccharide)
  • Novel shellfish pattern recognition receptor LRPP (Leucine-rich Repeat Protein) specifically combined with LPS (Lipopolysaccharide)
  • Novel shellfish pattern recognition receptor LRPP (Leucine-rich Repeat Protein) specifically combined with LPS (Lipopolysaccharide)

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Experimental program
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Embodiment 1

[0019] 1. RNA extraction

[0020] Total RNA was extracted using Trizol (Invitorgen), and the specific steps were as follows: (1) Take 50 mg of Pacific oyster tissue in a liquid nitrogen pre-cooled mortar, grind the tissue into powder, and transfer it to a centrifuge tube containing 1 ml of Trizol , pipetting, and mixing; (2) Add 200 μL of chloroform, shake vigorously for 40 seconds, and place at room temperature for 5 minutes; (3) Centrifuge at 12,000×g for 10 minutes at 4°C, transfer the supernatant to a new tube; (4) Add 0.5ml of isopropyl (5) Centrifuge at 12,000×g for 10 minutes at 4°C, and discard the supernatant; (6) Wash twice with 75% ethanol to precipitate; (7) Discard the supernatant, add 100 μL DEPC water to dissolve RNA.

[0021] 2. Construction of cDNA library and acquisition of full-length cDNA

[0022] 1) The SMART cDNA library construction Kit (Clontech) was used to construct the full-length cDNA library. The specific steps are as follows: (1) Use the RNA in ...

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Abstract

The invention discloses a novel shellfish pattern recognition receptor LRPP (Leucine-rich Repeat Protein) specifically combined with LPS (Lipopolysaccharide). The amino acid sequence of the shellfish pattern recognition receptor LRPP is as shown in SEQ ID NO.2. The LRPP is separated from crassostrea gigas for the first time, LPS can be specifically recognized, and the novel shellfish pattern recognition receptor LRPP can trigger later immunoreaction. The LRRP disclosed by the invention plays an important role in shellfish immunological recognition, can monitor the invasion situation of intracorporal gram-negative bacteria, and provides significant theory and practice basis for anti-disease breeding of shellfish.

Description

Technical field: [0001] The invention belongs to the molecular mechanism field of shellfish immune recognition and immune defense, in particular to a novel shellfish pattern recognition receptor LRRP specifically combined with LPS. Background technique: [0002] Shellfish occupies a pivotal position in the economic and social development of my country's coastal areas, and is one of the most important economic groups in the world, and has great potential for development. Pacific oyster (Crassostrea gigas) is a mollusc, bivalves, pearl oyster, oyster family animal, is an important marine biological resource, and is a globally distributed species. However, the lack of good species leads to low oyster immunity, disease outbreaks, and even mass mortality, which is the main bottleneck restricting the development of the industry. [0003] In the immune response of shellfish, the conserved structures of pathogenic microorganisms, such as lipopolysaccharide (LPS), peptidoglycan (PGN...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/46C12N15/12C12N15/70C12N1/21G01N33/68C12R1/19
CPCC07K14/705
Inventor 张扬张跃环李军喻子牛
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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