Novel shellfish opsonin DCP (DM9domain-containing protein) capable of promoting phagocytosis of lymphocytes

A blood lymphocyte, phagocytosis technology, applied in applications, bacteria, animal/human peptides, etc., can solve problems such as disease outbreaks, low immunity, and restricting industrial development.

Active Publication Date: 2014-05-28
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, with the large-scale breeding in recent years, its low immunity has led...

Method used

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  • Novel shellfish opsonin DCP (DM9domain-containing protein) capable of promoting phagocytosis of lymphocytes
  • Novel shellfish opsonin DCP (DM9domain-containing protein) capable of promoting phagocytosis of lymphocytes
  • Novel shellfish opsonin DCP (DM9domain-containing protein) capable of promoting phagocytosis of lymphocytes

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Embodiment 1

[0021] 1. RNA extraction

[0022] Total RNA was extracted using Trizol (Invitorgen), and the specific steps were as follows: (1) Take 50 mg of Pacific oyster in a liquid nitrogen pre-cooled mortar, grind the tissue into powder, and transfer it to a centrifuge tube containing 1 ml Trizol, Pipette and mix well; (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 isopropanol , mix well, and precipitate overnight at -80°C; (5) centrifuge at 4°C, 12,000×g for 10 min, discard the supernatant; (6) wash twice with 75% ethanol for precipitation; (7) discard the supernatant, add 100 μL DEPC water to dissolve the RNA .

[0023] 2. RACE (Rapid Amplification of cDNA Ends) and full-length cDNA acquisition

[0024] (1) Using the extracted RNA and GeneRacerTM kit (Invitrogen), construct the Pacific oyster RACE cDNA library; (2) Ob...

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Abstract

The invention discloses a novel shellfish opsonin DCP (DM9domain-containing protein) capable of promoting the phagocytosis of lymphocytes. The amino acid sequence of the opsonin DCP is shown in SEQ ID NO. 2. According to the invention, DCP is isolated from crassostrea gigas for the first time; function analysis and identification show that the DCP of the crassostrea gigas can be bound with surface molecules of vibrio parahaemolyticus, can greatly promote the phagocytosis of lymphocytes on the vibrio parahaemolyticus, and is a novel shellfish opsonin. As the DCP can greatly promote the phagocytosis of lymphocytes, an important theory and practice basis are provided for disease prevention and control of shellfish.

Description

Technical field: [0001] The invention belongs to the technical field of applied marine organisms, and in particular relates to a novel opsonin DCP for promoting the phagocytosis of blood lymphocytes by shellfish. Background technique: [0002] Shellfish is one of the most important economic groups in my country and plays an important role in the economic and social development of coastal areas. The Pacific oyster (Crassostrea gigas) belongs to the mollusk phylum, bivalve class, pearl oyster order, oyster family animal, and is a globally distributed economic shellfish. However, with the large-scale breeding in recent years, its low immunity has led to disease outbreaks, which have seriously restricted the development of the industry. [0003] Hemolymphocytes are the core immune organs of shellfish, and the strength of their phagocytic ability directly determines the level of their disease resistance. In oysters, there is a class of protein that can promote the phagocytosis ...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12C12N15/70C12N1/21A61K38/17A61P37/04
CPCA61K38/00C07K14/43504
Inventor 张扬徐凤姣张跃环李军喻子牛
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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