Recombinant serum amyloid A capable of enhancing immune response of crassostrea gigas, and preparation method thereof

A technology of immune response and serum starch, applied in the fields of biochemistry and molecular biology, can solve the problems such as undiscovered long oysters, no long oyster innate immune response level, etc., and achieve the effect of enhancing the immune response level

Inactive Publication Date: 2020-02-28
DALIAN OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there have been no reports on the physiological function of SAA in long oyster, and no drugs that can significantly promote the level of innate immune response in long oyster

Method used

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  • Recombinant serum amyloid A capable of enhancing immune response of crassostrea gigas, and preparation method thereof
  • Recombinant serum amyloid A capable of enhancing immune response of crassostrea gigas, and preparation method thereof
  • Recombinant serum amyloid A capable of enhancing immune response of crassostrea gigas, and preparation method thereof

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experiment example

[0048] 1. Recombinant protein r of the present invention C g Comparison of different induced expression of SAA

[0049] According to the preparation methods a, b, and c of the embodiment of the present invention, single clones were picked and inoculated in 200 mL of LB liquid medium at 220 rpm, and cultured at different temperatures (37 °C, 28 °C, 16 °C) to OD 600 = 0.6. Add different final concentrations (1.5 mmol L -1 , 1 mmol L -1 , 0.5 mmol L -1 ) with the inducer IPTG for 4 hours, and then centrifuged at 5000 rpm for 10 minutes at 4 °C to collect the bacteria and freeze them at -20 °C for future use. Take 1 mL of bacteria and add 80 μL of water and 20 μL of 5-fold protein loading buffer, boil at 100 °C for 10 minutes, centrifuge slightly, and detect the expression product by SDS-PAGE. The result is as figure 1 shown. figure 2 Middle A: Different culture temperature (37 ℃, 28 ℃, 16 ℃) and IPTG inducer concentration (1.5 mmolL -1 , 1 mmol L -1 , 0.5 mmol L -1...

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Abstract

The invention discloses recombinant serum amyloid A capable of enhancing immune response of crassostrea gigas. The recombinant serum amyloid A is characterized that the amino acid sequence is represented by SEQ ID NO. 1. Gene expression of cytokines CgIL17-1, CgIL17-5, and CgTNF in blood lymphocytes can be remarkably promoted inside and outside bodies of the crassostrea gigas, the immune responselevel of the crassostrea gigas can be enhanced, and the recombinant serum amyloid A has application value in preparation of novel immune preparations and other drugs for shellfishes, especially breeding shellfishes.

Description

technical field [0001] The invention belongs to the technical field of biochemistry and molecular biology, and in particular relates to a serum amyloid A capable of enhancing the immune response of oysters and a preparation method thereof. [0002] technical background [0003] long oysters ( Crassostrea gigas , C g ) is an important marine economic shellfish in my country. In recent years, various diseases caused by bacteria, fungi and viruses have continuously broken out in oyster culture groups, causing huge economic losses. Due to the lack of adaptive immune defense system, long oyster mainly relies on innate immune response to resist the infection of exogenous pathogenic microorganisms. The latest research shows that the long-term latent infection of the pathogen leads to the insufficiency of the innate immune response of oysters, which leads to the imbalance of homeostasis, which is one of the important reasons for the mass mortality of oysters. In mammals, serum a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K1/36C07K1/34C07K1/16C12N15/70A61K38/17A61P37/04
CPCC07K14/43504C12N15/70A61P37/04A61K38/00
Inventor 王玲玲王伟林吕晓静孔宁宋林生
Owner DALIAN OCEAN UNIV
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