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Recombinant lactococcus lactis and vaccine for Streptococcus agalactiae disease of Oreochromis sp.

A Lactococcus lactis, Streptococcus lactis technology, applied in vaccines, veterinary vaccines, recombinant DNA technology and other directions, can solve the problems of drug residue quality and safety, chemical drug resistance and other problems, to improve convenience and simple preparation process , the effect of reducing workload

Pending Publication Date: 2020-09-22
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chemical drug prevention and control is the most direct, convenient and convenient means of prevention and control, but long-term use of chemical drugs is prone to drug resistance, resulting in quality and safety problems caused by drug residues

Method used

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  • Recombinant lactococcus lactis and vaccine for Streptococcus agalactiae disease of Oreochromis sp.
  • Recombinant lactococcus lactis and vaccine for Streptococcus agalactiae disease of Oreochromis sp.
  • Recombinant lactococcus lactis and vaccine for Streptococcus agalactiae disease of Oreochromis sp.

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Construction of recombinant Lactococcus lactis based on the fbsA gene

[0026] 1. Construction of recombinant plasmid pNZ8148-fbsA

[0027] Design fbsA primers, and add 6×His tags at the end of the target gene primers (the part marked with a horizontal line in the sequence in Table 1 below), select the general primer of pNZ8148 as the identification primer, and the primers used in the experiment were synthesized in Guangzhou Aiji Biotechnology Co., Ltd. company, and the primer sequences are listed in Table 1.

[0028] Table 1

[0029]

[0030] Cultivate Streptococcus agalactiae WC1535 in BHI medium overnight at 37°C and 180r / min, use TaKaRaMiniBEST Bacteria Genomic DNA Extraction Kit to extract the genomic DNA of Streptococcus agalactiae, and use the genomic DNA of Streptococcus agalactiae as a template to amplify fbsA Gene. The reaction system is shown in Table 2.

[0031] Table 2

[0032]

[0033] PCR amplification program: 94°C for 10 min, 94°...

Embodiment 2

[0057] Example 2: Using recombinant Lactococcus lactis to prepare a vaccine and evaluate the protective effect of the vaccine

[0058] Select 350 healthy tilapias (body length 8 ± 1cm, body weight 14 ± 1g) from the tilapia breeding farm in Guangdong Province, and randomly divide them into 7 groups, 50 fish in each group, grouped as: recombinant Lactococcus lactis bacteria liquid group , recombinant Lactococcus lactis liquid + Essai GR01 PR adjuvant + feed group, adjuvant + feed group, L.lactis NZ9000 pNZ8148 bacterial liquid control group, L.lactisNZ9000 bacterial liquid control group and PBS blank control group.

[0059] Recombinant Lactococcus lactis was induced and cultured at 30°C in the presence of 10 ng / mL lactobacillus, the cells were collected from the induced expression culture, and diluted with PBS to prepare the bacterial solution for immunization.

[0060] In the experimental group involving recombinant Lactococcus lactis, the number of recombinant Lactococcus la...

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Abstract

The invention discloses recombinant lactococcus lactis based on the fbsA gene of Streptococcus agalactiae. The recombinant lactococcus lactis comprises the fbsA gene of Streptococcus agalactiae or segments of the fbsA gene, the fbsA gene or the segments of the fbsA gene preferably has (or have) a sequence shown in SEQ ID No. 1, the deposit number of the recombinant lactococcus lactis is CCTCC M 2020140, and expression of recombinant FbsA protein with a theoretical molecular weight of 34.6 kDa can be induced under the culture condition that niusin exists. The invention also provides a vaccine for the Streptococcus agalactiae disease of Oreochromis sp., and the vaccine is prepared from the recombinant lactococcus lactis, and is capable of producing protective antibodies in Oreochromis sp. after Oreochromis sp. is fed with the vaccine in an immune mode of oral gavage, so that Oreochromis sp. is assisted to resist infection of Streptococcus agalactiae effectively.

Description

technical field [0001] The invention belongs to the technical field of prevention and control of aquatic organism diseases, and in particular relates to a recombinant Lactococcus lactis constructed based on the fbsA gene of Streptococcus agalactiae and a tilapia Streptococcus agalactiasis vaccine prepared by using the recombinant Lactococcus lactis. Background technique [0002] Tilapia (Oreochromis sp.) is one of the main freshwater aquaculture species in southern my country. It has the advantages of miscellaneous food habits, wide adaptability, strong reproductive ability, and fast growth rate. In recent years, the outbreak of tilapia disease has caused huge losses to the tilapia industry, and the trend has become more serious, hindering the healthy development of the tilapia industry. One of the main pathogens of tilapia diseases is Streptococcus agalactiae. [0003] The main causes of tilapia streptococcosis agalactiae are poor water quality, high breeding density, and ...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/31C12N15/74A61K39/09A61P31/04C12R1/01
CPCC07K14/315C12N15/74A61K39/092A61P31/04A61K2039/523A61K2039/552A61K2039/542
Inventor 赖迎迢石存斌陶家发袁玉梅孙承文巩华
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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