Pseudomonas proteus znuA gene stable silencing strain and application

A technology of Pseudomonas mutans and bacterial strains, applied in the field of microbiology

Active Publication Date: 2021-04-09
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

To date, no information on Pseudomonas mutans wxya Reports on the effect of genes on the host

Method used

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  • Pseudomonas proteus znuA gene stable silencing strain and application
  • Pseudomonas proteus znuA gene stable silencing strain and application
  • Pseudomonas proteus znuA gene stable silencing strain and application

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Embodiment Construction

[0024] Pseudomonas mutans provided by the embodiments of the present invention wxya The method for the construction of gene stable and efficient silencing strain and its function research comprises the following steps:

[0025] S101: Using comparative transcriptomics technology to analyze the gene expression of Pseudomonas mutans in the spleen of grouper, it was found that wxya Gene-specific high expression will lock the target wxya Gene;

[0026] S102: For wxya The gene sequence was designed and synthesized using the online shRNA design tool of Thermo-fisher Scientific (http: / / rnaidesigner.thermofisher.com / rnaiexpress / ), and 5 pairs of shRNA primers were connected into pCM130 / tac to construct the recombinant vector , and then each recombinant vector was electroporated into Pseudomonas mutans competent cells, and Pseudomonas mutans was successfully constructed wxyaGene stable silencing strains; using qRT-PCR technology, the primers are F: 5'-AGGTGATCTTCTCGGAGATGGA-3',...

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Abstract

The invention discloses a pseudomonas proteus znuA gene stable silencing strain and application. The pseudomonas proteus znuA gene stable silencing strain is constructed through a gene silencing technology, then the pathogenicity of the znuA gene stable silencing strain to epinephelus coioides is determined through an artificial infection experiment, transcriptome data of spleens of the epinephelus coioides infected by a wild strain and the znuA gene stable silencing strain is analyzed by utilizing a dual RNA-seq technology, meanwhile, transcriptome data of hosts and pathogenic bacteria is researched, the function of a znuA gene in interaction of pseudomonas proteus and the epinephelus coioides is discussed from the transcriptome level, and then the influence of the znuA gene on the toxicity of the pseudomonas proteus is disclosed. The constructed strain has the advantages that the pathogenicity to the epinephelus coioides is remarkably reduced, and the transcriptome data of the pseudomonas proteus and the epinephelus coioides in the infected spleens can be remarkably changed, so that the strain can be used for researching the pathogenic mechanism of the pseudomonas proteus.

Description

technical field [0001] The invention belongs to the technical field of microbes, and more specifically relates to a bacterial strain valuable for studying the pathogenic mechanism of Pseudomonas mutans. Background technique [0002] Pseudomonas mutans ( Pseudomonas plecoglossicida ) is the pathogenic bacterium of "visceral white spot disease" of mariculture fish such as large yellow croaker and oblique grouper, and the direct economic loss caused by it exceeds 100 million yuan every year. [0003] Zn(II) is an indispensable substance for bacteria and exists in many proteins and enzymes involved in nucleic acid metabolism processes, as well as some ribosomal proteins. In the absence of zinc, ZnuABC and ZupT are the most conserved zinc transport systems in many bacteria, and inactivation of ZnuABC reduces the virulence and colonization ability of bacteria. The ZnuABC transporter consists of ZnuA, ZnuB and ZnuC. ZnuA is a soluble periplasmic component that captures Zn(II) an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/113C12R1/38
CPCC07K14/21C12N15/113C12N2310/14
Inventor 赵玲敏王路英鄢庆枇何乐黄力行覃映雪徐晓津马英郑江
Owner JIMEI UNIV
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