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

A technology of mutagenic strain and leech vibrio, applied in the field of microbial mutagenesis and breeding, can solve the problems of low mutation rate and difficult recovery, and achieve the effects of improving the lysis ability, enhancing the lysis ability and broadening the scope of application

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

Traditional mutagenesis breeding is mainly based on ultraviolet mutagenesis, but its mutation rate is not high and requires multiple accumulated mutations, which is very easy to recover; and Co 60 Irradiation mutagenesis can obtain a higher mutation rate and a wider mutation spectrum, and it is not easy to recover
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 wide lytic spectrum and its application
  • A mutant strain of Bdellovibrio with strong lytic performance and wide lytic spectrum and its application
  • A mutant strain of Bdellovibrio with strong lytic performance and wide lytic spectrum and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Co 60 once irradiated

[0067] (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 20‰, dispersed and mixed evenly, then transferred into a centrifuge tube to obtain the host bacteria concentrate, bagged, stored at 4°C, and used for later use;

[0068] (2) Preparation of 28 strains of host bacteria concentrate: Take one of each of the 28 strains of ampoule-preserved strains, wipe the mouth of the bottle with alcohol cotton, gently b...

Embodiment 2

[0082] Example 2 Co 60 secondary irradiation

[0083] (1) Host culture: same as step (1) of Example 1.

[0084] (2) Preparation of concentrated solution of 28 strains of host bacteria: same as step (2) of Example 1.

[0085] (3) BDE-1F1 seed liquid culture: Referring to step (3) of Example 1, pick phage plaques from the double-layer plate of the mutant strain BDE-1F1.

[0086] (4) Preparation of irradiated samples: take the BDE-1F1 seed solution in step (3), refer to step (4) of Example 1, and increase the irradiation dose to 5Gy / min×100min.

[0087] (5) Screening of mutagenic strains: Refer to step (5) of Example 1 to obtain the seed solution of the secondary mutagenic strain BDE-1F2.

[0088] (6) Detection of the ability of the mutant strain to lyse pathogenic Vibrio: refer to step (6) of Example 1.

[0089] Double-layer plate diagram of Bdellovibrio mutagen BDE-1F2 lysing Vibrio alginolyticus 2, see image 3 .

[0090] Two-layer plate diagram of Serratia figum 15 lyse...

Embodiment 3

[0103] Example 3 Co 60 Comparison of the ability of lysing positive bacteria between the mutant strain and the wild strain after the first and second irradiation

[0104] (1) Preparation of Gram-positive bacteria concentrate: Take one of the four strains of Gram-positive bacteria preserved in ampoules, wipe the mouth of the bottle with alcohol cotton, gently break the mouth of the bottle, and absorb 1 mL of NB liquid medium Pipette the powder until dissolved, transfer it to 50mL bottled NB liquid medium, incubate at 37°C, 200r / min for 8h, put it in a refrigerated centrifuge at 6000rpm, 4°C for 10min, then discard 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 bacteria, bag it, store at 4°C, and set aside;

[0105] (2) Detection of the ability of the mutagenic strain to lyse Gram-positive bacteria: refer to step (6) of Example 1.

[0106] Compare Co 60 The number and size ...

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Abstract

The invention discloses a leech vibrio mutant strain with strong lysis performance and wide lysis spectrum and its application. The leechvibrio mutant strain is Bdellovibrio sp.BDE-1F2, which was deposited in the Guangdong Provincial Microbial Culture Collection Center on November 22, 2018, with the deposit number: GDMCC NO: 60491. The present invention utilizes Co 60 Irradiation-induced mutation method to enhance the lysis performance of Hirudovibrio, effectively improve the lytic ability of Hirudovibrio against Gram-negative and positive pathogenic bacteria and / or potential pathogenic bacteria, and broaden the application scope of Hirudovibrio , to better meet the needs of production practice. The lysis performance of the leech vibrio mutant strain of the present invention is tested to confirm that it can lyse four gram-negative bacteria that cannot be lysed by the original, so that the total lysis rate of 28 indicator bacteria reaches 100%, and the lysis of Gram-negative bacteria is enhanced. The lysis ability of positive bacteria provides a method basis for the enhancement of the lysis performance of Hirudovibrio.

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 wide lytic spectrum and its application. Background technique [0002] Bdellovibrio is a class of small bacteria capable of lysing other bacteria. It can parasitize and lyse Gram-negative bacteria of most families and genera. Its lysing effect on some pathogenic bacteria makes it of great value. At present, most Bdellovibrio cultures use 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 and adhesion of Bdellovibri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P1/00C12R1/01
CPCC12N1/20C12N1/205C12R2001/01
Inventor 蔡俊鹏曹青青
Owner SOUTH CHINA UNIV OF TECH
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