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Edwardsiella tarda outer membrane protein vaccine and its preparation and application

A technology for Edwards tarda and outer membrane protein, which is applied in the field of Edwardian tarda outer membrane protein vaccine and its preparation, can solve problems such as not yet applied on a large scale, and achieve the effect of promoting immune activity

Active Publication Date: 2022-06-24
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In response to the ravages of Edwardsiella tarda, South Korea has invented a commercialized vaccine based on inactivated whole bacteria. In my country, due to the late start of related research, so far, only one attenuated vaccine of Edwardsiella tarda has obtained the national new veterinary drug Certificates, but not yet used on a large scale

Method used

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  • Edwardsiella tarda outer membrane protein vaccine and its preparation and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The E. lentus outer membrane protein vaccine antigen is shown in the amino acid sequence in SEQ ID No. 1 of the sequence table, and the amino acid sequence is a sequence on the E. Sequence, the base sequence except the signal peptide part was obtained by gene synthesis as the antigen sequence (synthesis was completed by Sangon Bioengineering Co., Ltd. (Shanghai)), and the translated amino acid sequence was as follows.

[0027] SEQ ID No. 1:

[0028] MSSGDSTITLNYIQQSNGQVEKDLAGFKQITDQFIGSEHFGASTAPYRDAEGVALSYRYEFTDCWGVIGRLSYTGLRRGMQIRRGHNYGPGVPLVLVDGRSRSQRWGIMAGPSYRVTDNLSLYGLAGASVDRLSWHIQVDDGANDVLGTALHTADQQLTRVSMAYAAGVQLNAGGYVLDFSYTGVGGDDRSHGFLVGVGLIF

[0029] (a) Sequence features:

[0030] Length: 201

[0031] Type: amino acid sequence

[0032] ·Chain type: single chain

[0033] Topology: Linear

[0034] (b) Molecular type: protein

[0035] (c) Assumption: No

[0036] (d) Antonym: No

[0037] (e) Original source: Edwardsiella lentus TX1

[0038] (f) Specificity ...

Embodiment 2

[0041] Construction method of Edwardsiella lentus vaccine expression vector

[0042] Construction of the recombinant plasmid pETOmp1: The antigenic base sequence of the Edwardella lentus outer membrane protein vaccine synthesized by the above-mentioned Sangon Bioengineering Co., Ltd. (Shanghai) was used as a template, and it was connected to the commercial plasmid pET28a by homologous recombination. , the recombinant plasmid pETOmp1 was obtained. The LB liquid medium is used for culturing Escherichia coli, and the components are in percentage by weight: 1.0% peptone, 0.5% yeast powder, 1.0% sodium chloride, 97.5% distilled water;

Embodiment example 3

[0044] Inducible expression and purification of vaccine proteins

[0045] Vaccine protein rOmp1 Et Inducible expression and purification: The plasmid pETOmp1 was transformed into Escherichia coli BL21(DE3) (Beijing, Quanshijin Biotechnology Co., Ltd.), and the bacterial solution was cultured on LB solid medium containing kanamycin (50 μg / ml) After 18-24 hours, a positive clone BL21 / pETOmp1 was screened and obtained. BL21 / pETOmp1 was cultured overnight in LB liquid medium containing kanamycin (50 μg / ml). Take 5ml of the bacterial liquid after overnight culture, add it to 500ml of fresh LB liquid medium containing kanamycin (50μg / ml), shake and cultivate to OD at 37°C in a shaker with a rotating speed of 180rpm. 600 After ≈0.5, IPTG was added at a final concentration of 1 mM, and the culture was continued for 16 hours with shaking at 16°C and 180 rpm. Centrifuge at 6000g at 4°C for 10 minutes, and collect the bacterial cells cultured under the above conditions. Add 5 ml of l...

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Abstract

The invention relates to the field of molecular vaccinology, in particular to an Edwardsiella tarda outer membrane protein vaccine and its preparation method and application. The outer membrane protein vaccine contains the Edwardsiella tarda outer membrane protein vaccine antigen shown by the amino acid in SEQ ID No.1. The recombinant protein vaccine constructed by the invention can protect the flounder against the infection of Edwardsiella tarda, induce the production of specific antibodies, and the protein can significantly promote the immune activity of fish immune cells.

Description

technical field [0001] The invention relates to the field of molecular vaccinology, in particular to an Edwardsiella lentus outer membrane protein vaccine and a preparation method and application thereof. Background technique [0002] Edwardsiella tarda is a Gram-negative pathogen that infects a variety of animals, including fish and mammals. Edwardsiella tarda can cause Edwardsiella tarda disease in mariculture fish, causing serious economic losses to my country's aquaculture industry. There are currently reports of mass mortality from E. lentus in a variety of farmed fish. In response to the raging of Edwardella tarda, a commercial vaccine based on inactivated whole bacteria has been invented in South Korea. In my country, due to the late start of related research, so far, only one attenuated Edwardella tarda vaccine has obtained the national new veterinary drug certificate, but not yet widely used. Therefore, it is urgent to develop other types of vaccines against Edwar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/02A61K39/39A61P31/04C07K14/24C12N15/70
CPCA61K39/025A61K39/39A61P31/04C07K14/24C12N15/70A61K2039/552A61K2039/55505
Inventor 李墨非金成东孙黎
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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