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Method for improving formation of citrobacter werkmanii biofilm

A technology of citrobacter and biofilm, applied in the field of genetic engineering, can solve the problems of limited biofilm formation ability and poor environmental adaptability of Citrobacter weimanii, and achieve the improvement of biofilm formation, drug resistance level and drug resistance , Broaden the effect of application scenarios and scope

Active Publication Date: 2020-09-01
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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Problems solved by technology

[0004] The purpose of the present invention is to provide a kind of method that improves the biofilm formation of Citrobacter wilsonii for the limited biofilm formation ability of Citrobacter wilsonii in the prior art, which can improve the quality of Citrobacter wilsonii lemon. The formation of acid bacilli biofilm can improve the resistance to fungicides, thereby improving its application potential in environmental governance and other aspects

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  • Method for improving formation of citrobacter werkmanii biofilm
  • Method for improving formation of citrobacter werkmanii biofilm

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Embodiment Construction

[0019] The following examples are to further illustrate the present invention, rather than limit the present invention.

[0020] The Citrobacter welkii used in the following examples is Citrobacter welmannii BF-6.

[0021] The primers used in the following examples are as follows:

[0022] ACGCCTGTCTCAAGCGGTTTTC(ompA-identify-F);

[0023] AAGAAGCATCGTGAGGGGGAAT(ompA-identify-R).

[0024] 1. Construction of ompA knockout vector

[0025] First, the upstream and downstream sequences (upstream 1005bp (corresponding to the 9th to 1013th bases of SEQ ID NO.2) of the ompA gene (its nucleotide sequence is shown in SEQ ID NO.1) and downstream 876bp (corresponding to the 1014th to 1889th bases of SEQ ID NO.2)) integrated together (without ompA gene itself sequence), then add BglII restriction site (GAAGATCT) at the front of the integrated sequence, followed by Add the SpeI restriction site (ACTAGTCC) and the protective base to the end, and entrust Shanghai Meiji Biomedical Technolog...

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Abstract

The invention discloses a method for improving the formation of a citrobacter werkmanii biofilm and provides a method for improving the biofilm forming ability of a strain by knocking out an outer membrane protein ompA gene of citrobacter werkmanii. A knockout vector is constructed by using a plasmid pYG4, and an ompA gene is knocked out by using a gene homologous recombination principle, so thatdelta ompA is obtained. The delta ompA character is analyzed through crystal violet staining and other methods, and the result shows that compared with wild citrobacter werkmanii, the biofilm formingability of delta ompA is improved by four times or above, and the drug resistance of delta ompA to isothiazolinone bactericides is also correspondingly improved. According to the ompA knockout strainconstructed by the invention, the formation amount and the drug resistance level of a citrobacter welchii biofilm can be effectively improved, so that the application scenarios and ranges of citrobacter werkmanii for heavy metal ion adsorption, construction of cell protein synthesis factories and the like are broadened.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a method for improving the biofilm formation of Citrobacter wilsonii. Background technique [0002] Citrobacter welmanii, a Gram-negative bacterium, is able to utilize L-arabinose, maltose, rhamnose, trehalose, D-xylose, D-mannose, and D-sorbose through peri-flagellar movement Sugar alcohols, etc. are single carbon sources, and the reaction is positive for malonate and m-hydroxybenzoate, which are usually found in water, soil, animal and human intestines. Based on the above physiological characteristics of Citrobacter weimanii, it has been applied in the field of environmental governance, such as Citrobacter weimani itself, especially the biofilm formed by it, has a good ability to accumulate heavy metals and is used for heavy metal pollution in the environment However, due to its limited ability to form a biofilm, even if a biofilm is formed, its environmental ada...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12N1/21C12Q1/18C12R1/01
CPCC07K14/24C12N15/74C12Q1/18
Inventor 周刚王颖思彭红施庆珊谢小保李素娟孙廷丽沈鹏飞
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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