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Pseudomonas putida ND6 coupled protein complex dotM gene deletion mutant bacterium as well as construction method and determination method thereof

A technology of Pseudomonas putida and gene deletion, applied in the biological field, can solve the problems of secondary gene pollution, ecological environment imbalance, unfavorable naphthalene degradation, etc.

Active Publication Date: 2021-02-09
XI AN JIAOTONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

During the practical application of Pseudomonas putida ND6, it was found that the conjugative transfer system has no selectivity, not only can transfer the naphthalene degradation ability of ND6 to other strains, but also can transfer other abilities of ND6 to other strains, which is easy to trigger gene Secondary pollution, resulting in an imbalance in the ecological environment, is not conducive to the degradation of naphthalene

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  • Pseudomonas putida ND6 coupled protein complex dotM gene deletion mutant bacterium as well as construction method and determination method thereof
  • Pseudomonas putida ND6 coupled protein complex dotM gene deletion mutant bacterium as well as construction method and determination method thereof
  • Pseudomonas putida ND6 coupled protein complex dotM gene deletion mutant bacterium as well as construction method and determination method thereof

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Embodiment Construction

[0070] Below in conjunction with accompanying drawing and specific embodiment content of the present invention is described in further detail:

[0071] The dotM gene deletion mutant of the Pseudomonas putida ND6 coupling protein complex containing a resistance marker has a biological deposit number of CGMCC No. 20500, and the dotM gene has a deletion of nucleotides 20751-21419.

[0072] At the same time, the present invention also provides a method for constructing the above-mentioned Pseudomonas putida ND6 coupling protein complex dotM gene deletion mutant containing a resistance marker, which is special in that it includes the following steps:

[0073] 1) Cloning of genes and construction of recombinant suicide plasmids

[0074] 1.1) Referring to the dotM gene sequence in GenBank, use the ND6 genome as a template to amplify in vitro to obtain the upstream and downstream fragments of the dotM gene. The specific steps are:

[0075] A) ND6 Genomic DNA Extraction

[0076] A1) ...

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Abstract

The invention provides a pseudomonas putida ND6 coupled protein complex dotM gene deletion mutant bacterium containing a resistance marker as well as a construction method and a determination method of the mutant bacterium, and provides a basis for further researching a conjugal transfer system. The biological preservation number of the pseudomonas putida ND6 coupled protein complex dotM gene deletion mutant bacterium containing the resistance marker is CGMCC No.20500.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a Pseudomonas putida ND6 coupling protein complex dotM gene deletion mutant containing a resistance marker and a construction method and a measurement method thereof. Background technique [0002] As one of the polycyclic aromatic hydrocarbon pollutants, naphthalene widely exists in the soil environment. Due to its water solubility, it is very easy to diffuse. Blood health poses certain threats. [0003] At present, there are many kinds of naphthalene-degrading bacteria. During the biodegradation process of naphthalene, most of the naphthalene-degrading genes exist on plasmids other than independent chromosomes, which are called naphthalene-degrading plasmids. These plasmids are usually capable of conjugative transfer. horizontal gene transfer. Pseudomonas putida ND6 is a kind of naphthalene-degrading bacteria, which contains two giant plasmids: pND6-1 and pND6-2, and the f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/78C12N15/65C12Q1/04B09C1/10C12R1/40
CPCC07K14/21C12N15/78C12N15/65C12Q1/04B09C1/10G01N2333/21
Inventor 李珊珊杜丹徐浩延卫
Owner XI AN JIAOTONG UNIV