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Method for in-situ fluorescent labeling of geobacillus

A technology of fluorescent labeling and geobacteria, applied in the field of microorganisms, can solve the problems of in situ fluorescent labeling, limitation, and short half-life of bacillus

Active Publication Date: 2020-07-14
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

However, the synthesis of its chromophore requires oxygen molecules as an essential cofactor, so it cannot be applied in the anaerobic bacteria Geobacter
In addition, the half-life of GFP is short, which also limits its long-term in situ observation of biofilm formation.
Therefore, finding a fluorescent probe that can emit light under anaerobic conditions and has a long half-life and its labeling method is an effective strategy to solve the current in situ fluorescent labeling of Geobacter.

Method used

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  • Method for in-situ fluorescent labeling of geobacillus
  • Method for in-situ fluorescent labeling of geobacillus
  • Method for in-situ fluorescent labeling of geobacillus

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Embodiment

[0037] Strains involved in this example: (1) Escherichia coli strain DH 5α chemically competent cells (purchased from Beijing Quanshijin Biotechnology Co., Ltd.) were used for gene cloning and plasmid amplification. (2) Geobactersulfurreducens PCA (purchased from the American Type Culture Collection, strain number ATCC-51573) was used to construct fluorescent protein-labeled strains.

[0038] The NBAF substratum formulation involved in cultivating bacterial strains in the present embodiment is as follows:

[0039]

[0040] *42g KH per liter of 100X NB salt solution 2 PO 4 , 22g K 2 HPO 4 , 20g NH 4 Cl, 38g KCl, 36g NaCl.

[0041] **Contains 2.14g NTA per liter of NB mineral solution; 0.1g MnCl 2 *4H 2 O; 0.3g FeSO 4 *7H 2 O; 0.17g CoCl 2 *6H 2 O; 0.2g ZnSO 4 *7H 2 O; 0.03g CuCl 2 *2H 2 O; 0.005g AlK(SO 4 ) 2 *12H 2 O; 0.005g H 3 BO 3 ;0.09g Na 2MnO 4 *2H 2 O; 0.11 g NiSO 4 *6H 2 O; 0.02g Na 2 WO 4 *2H 2 O.

[0042] ***Per liter of DL vitamin sol...

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Abstract

The invention provides a method for in-situ fluorescent labeling of geobacillus. The method includes transferring a PpFbFP fluorescent protein gene into geobacillus. The PpFbFP fluorescent protein label is not influenced by oxygen, can make the labeled geobacillus strain fluoresce under anaerobic conditions, and has a long half-life, and can realize long-term real-time observation of movement trajectory, film forming process and group behavior of the geobacillus.

Description

technical field [0001] The invention relates to a method for in-situ fluorescent labeling of Geobacter, belonging to the technical field of microorganisms. Background technique [0002] Geobacter is the electroactive bacteria with the largest number and widest distribution in the natural environment, and also the electroactive model strain with the most extensive research and the strongest electron transfer activity. It is widely distributed in freshwater sediments, groundwater sediments polluted by organic matter and heavy metals It has three functions of iron reduction, humus reduction and electricity generation. The electroactive biofilm formed by Geobacter has important application value in environmental remediation and bioengineering. It can produce electricity or other chemicals while treating pollutants, which helps pollution remediation technology to low input, low energy consumption, Transformation in the direction of low pollution. Using in situ fluorescent prote...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12Q1/04C12N15/74C12N15/66C12R1/01
CPCC07K14/43595C12N15/74C12Q1/04
Inventor 刘星靖宪月高鹏宇刘璐周顺桂
Owner FUJIAN AGRI & FORESTRY UNIV
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