Bdellovibrio sp. induced mutation strain with strong cracking performance and application thereof

A technology of leech vibrio and mutagenic strains is applied in the field of microbial mutation breeding to achieve the effects of broadening the application range, improving the lysis ability and enhancing the lysis ability

Active Publication Date: 2020-07-03
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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  • Bdellovibrio sp. induced mutation strain with strong cracking performance and application thereof
  • Bdellovibrio sp. induced mutation strain with strong cracking performance and application thereof
  • Bdellovibrio sp. induced mutation strain with strong cracking performance and application thereof

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 present invention discloses a bdellovibrio sp. induced mutation strain with the strong cracking performance and an application thereof, and belongs to the technical field of microorganism inducedmutation breeding. The bdellovibrio sp. induced mutation strain is Bdellovibrio sp. BDN-1F2, which is collected in the Guangdong Microbial Culture Collection center in June 11, 2018 and has the collection number of GDMCC NO:60385. The cracking performance of bdellovibrio sp. is enhanced by a Co60 irradiation induced mutation method, so that the cracking capability of the bdellovibrio sp. to gram-negative and positive pathogenic bacteria is effectively improved, the application range of the bdellovibrio sp. is expanded, and the requirements of production practice are met. The cracking performance of the bdellovibrio sp. induced mutation strain is checked to prove that two strains of bdellovibrio sp. which cannot be cracked can be cracked, and the cracking capability of the gram-positive bacteria is enhanced, so that a method basis is provided for separation and performance enhancement of more bdellovibrio sp. with the positive bacteria as hosts.

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