A mutant strain of Bdellovibrio with strong lytic performance and its application

A technology of Bdellovibrio and mutagenic strains, applied in the field of microbial mutation breeding, to achieve the effect of broadening the application range, enhancing the lysis ability, and improving the lysis ability

Active Publication Date: 2021-09-21
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But through Co 60 There is no report on radiation mutagenesis to enhance the lytic performance of Bdellovibrio

Method used

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  • A mutant strain of Bdellovibrio with strong lytic performance and its application
  • A mutant strain of Bdellovibrio with strong lytic performance and its application
  • A mutant strain of Bdellovibrio with strong lytic performance and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1Co60

[0063] Example 1Co 60 once irradiated

[0064] (1) Host culture: Take the newly cultivated host bacteria Bacillus subtilis GIM1.136, burn the mouth of the bottle under the outer flame of the alcohol lamp in the aseptic operation table, absorb 3mL of the host bacteria seed liquid and transfer it into a 250mL bottle of NB liquid In the culture medium, put the inoculated host bacteria in a constant temperature shaker at about 31°C for about 15 hours, then put them in a refrigerated centrifuge at 6000rpm and centrifuge at 4°C for 10min, then discard the supernatant, add to each tube 1mL of sterile distilled water with a salinity of 15‰, dispersed and mixed evenly, then transferred into a centrifuge tube to obtain the concentrated solution of host bacteria, bagged, stored at 4°C, and used for later use;

[0065] (2) Preparation of concentrated solution of Vibrio alginolyticus and Vibrio parahaemolyticus: Take one of the two strains of Vibrio preserved in ampoules, wipe the mouth o...

Embodiment 2

[0078] Example 2Co 60 secondary irradiation

[0079] (1) Host culture: same as step (1) of Example 1.

[0080] (2) Preparation of Vibrio alginolyticus and Vibrio parahaemolyticus concentrate: same as step (2) of Example 1.

[0081] (3) BDN-1F1 seed solution culture: Referring to step (3) of Example 1, pick phage plaques from the double-layer plate of the mutant strain BDN-1F1.

[0082] (4) Preparation of irradiated samples: take the BDN-1F1 seed solution in step (3), refer to step (4) of Example 1, and increase the irradiation dose to 5Gy / min×100min.

[0083] (5) Screening of mutagenic strains: Refer to step (5) of Example 1 to obtain the seed liquid of the secondary mutagenic strain BDN-1F2, and store it at 4°C for future use.

[0084] (6) Detection of the ability of the mutant strain to lyse pathogenic Vibrio: refer to step (6) of Example 1.

[0085] The double-layer plate diagram of the lysis of Vibrio parahaemolyticus by BDN-1F2 mutant strain of Bdellovibrio, see ima...

Embodiment 3

[0100] Example 3Co 60 Comparison of the ability of lysing positive bacteria between the mutant strain and the wild strain after the first and second irradiation

[0101] (1) Preparation of Gram-positive host bacteria concentrate: Take one of the four strains of Gram-positive host bacteria preserved in ampoules, wipe the mouth of the bottle with alcohol cotton, gently break the mouth of the bottle, and absorb 1mL of NB liquid The culture medium was pipetted until the powder was dissolved, and transferred to a 50mL bottled NB liquid medium, cultured at 37°C, 200r / min for 8h, placed in a refrigerated centrifuge, centrifuged at 6000rpm, 4°C for 10min, and then discarded the supernatant , add 1mL DNB liquid medium to each tube, mix well and transfer to a centrifuge tube to obtain a concentrated solution of Gram-positive host bacteria, bag it, store it at 4°C, and set aside;

[0102] (2) Detection of the ability of the mutant strain to lyse Gram-positive host bacteria: refer to ste...

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Abstract

The invention discloses a Bdellovibrio mutant strain with strong cracking performance and application thereof, belonging to the technical field of microbial mutation breeding. The Bdellovibrio mutagen strain is Bdellovibrio sp.BDN‑1F2, which was deposited in the Guangdong Microbial Culture Collection Center on June 11, 2018, with a preservation number of GDMCC NO: 60385. The present invention utilizes Co 60 Irradiation-induced mutation method to enhance the lysis performance of Bdellovibrio, effectively improves the lysis ability of Bdellovibrio to Gram-negative and positive pathogenic bacteria, broadens the application range of Bdellovibrio, and better meets the production practice needs. The lysing performance of the Bdellovibrio mutagenic strain of the present invention was tested to confirm that it can lyse two strains of pathogenic Vibrio that could not be lysed originally, and the ability to lyse Gram-positive bacteria was enhanced, and more leeches with positive bacteria as hosts could be lysed. Vibrio isolation and performance enhancement provide the methodological basis.

Description

technical field [0001] The invention belongs to the technical field of microbial mutation breeding, in particular to Co 60 The application of radiation mutagenesis in enhancing the lytic performance of a strain of Bdellovibrio with Bacillus subtilis as a parasitic host, especially relates to a mutagenic strain of Bdellovibrio with strong lytic performance and its application. Background technique [0002] Bdellovibrio is a class of small bacteria that can lyse other bacteria. It can parasitize and lyse Gram-negative bacteria of most families and genera. Its lytic effect on some pathogenic bacteria makes it of great value. Therefore, most Bdellovibrio cultures choose Gram-negative bacteria as hosts, such as Escherichia coli. [0003] The growth and reproduction of Bdellovibrio is also affected by various factors, including the host. First of all, there must be a sufficient number of host bacteria to ensure the collision, adhesion and invasion of Bdellovibrio and the host ba...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N13/00A61K35/741A61P31/04C12R1/01
CPCA61K35/74A61P31/04C12N1/20C12N13/00C12N1/205C12R2001/01
Inventor 蔡俊鹏陈丹
Owner SOUTH CHINA UNIV OF TECH
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