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Genetic engineering strain capable of highly producing phenazine-1-carboxylic acid, and construction method and applications thereof

A technology of genetically engineered strains and construction methods, applied in the field of genetically engineered strains with high yields of phenazine-1-carboxylic acid and its construction, capable of solving problems such as harsh conditions and unsuitable strains for production

Active Publication Date: 2020-12-25
QILU UNIV OF TECH
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Although there are chemical methods for the production of phenazine-1-carboxylic acid at present, the conditions are harsh, and toxic and harmful substances are released to the environment, so it is mainly produced by Pseudomonas aeruginosa in Pseudomonas, but Pseudomonas aeruginosa is It is a relatively common opportunistic pathogen in hospitals at present, and it is not suitable for promotion and application as a production strain

Method used

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  • Genetic engineering strain capable of highly producing phenazine-1-carboxylic acid, and construction method and applications thereof
  • Genetic engineering strain capable of highly producing phenazine-1-carboxylic acid, and construction method and applications thereof
  • Genetic engineering strain capable of highly producing phenazine-1-carboxylic acid, and construction method and applications thereof

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Experimental program
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Embodiment

[0072] 1. Inoculate Pseudomonas aeruginosa Qlu-1 into KB (A + ) culture medium, 30°C shaker, 180rpm extraction shaker and culture overnight, use the genome extraction kit to extract the genome of Qlu-1, and store it at -20°C for future use.

[0073] 2. Search the phzO gene and its upstream and downstream sequences in the sequenced Qlu-1 genome data, use the Qlu-1 strain genome as a template, and use phzO-F1 / phzO-R1, phzO-F2 / phzO-R2 as primers respectively Amplify the upstream fragment phzO-U of the phzO gene and the downstream fragment phzO-D; use phzO-U and phzO-D as templates, and phzO-F1 / phzO-R1 as templates to amplify the upstream and downstream fusion fragment phzO-UD of the phzO gene.

[0074] 3. The fusion fragment phzO-UD was ligated with the knockout plasmid pk18moBsacB to construct the recombinant plasmid pk18-phzO-UD.

[0075] 4. The recombinant plasmid pk18-phzO-UD was introduced into Escherichia coli S17-1(λ) by heat shock transformation.

[0076] 5. Escherichia...

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Abstract

The invention provides a genetic engineering strain capable of highly producing phenazine-1-carboxylic acid, and a construction method and applications thereof, and belongs to the technical field of genetic engineering. Relatively safe pseudomonas chlororaphis Qlu-1 is taken as an original strain; phzO genes,binary regulation related genes and central carbon metabolism pathway regulation genes areknocked out in the Qlu-1 strain genome through a genetic engineering technology; and the corresponding genetic engineering strain is constructed, so that the yield of the phenazine-1-carboxylic acidcan be greatly increased, and the strain can be more effectively used for biological control. Through experimental verification, the genetic engineering strain QPCA-7 has excellent effects; PCA yieldcan reach up to 7854 mg / L; good growth performance can be achieved; and thus, the strain has good practical application values.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a genetic engineering strain with high yield of phenazine-1-carboxylic acid and its construction method and application. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Chemical pesticide pollution is serious and unsustainable. Biological pesticides are a good substitute. Compared with chemical pesticides, biological pesticides have obvious advantages such as low toxicity, environmental friendliness and resistance to pesticides, and have attracted more and more people. Many scholars have paid attention to it and gradually applied it to the practice of b...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/78C12P17/12C12R1/38
CPCC07K14/21C12N15/78C12P17/12
Inventor 刘开泉李玲王瑞明王腾飞李丕武
Owner QILU UNIV OF TECH
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