DMSO preservation method of halophilic bacterial phage

A preservation method and phage technology, which is applied in the field of biochemistry, can solve the problems of poor preservation method effect, etc., and achieve the effect of long preservation time, good effect and simple preservation conditions

Pending Publication Date: 2021-12-14
YUNCHENG UNIVERISTY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the deficiencies in the prior art, the object of the present invention is to provide a DMSO preservation method for halophilic bacteriophages, which preserves the phages of Virgibacillus sp.SK39 isolated and screened from Yuncheng Salt Lake, and provides the most effective Aiming at the preservation method of the isolated potent phage this time, the problem of poor effect of the current preservation method is solved

Method used

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Embodiment 1

[0020] A DMSO preservation method for halophilic bacteriophage, comprising the steps of:

[0021] (1) Inoculate the halophilic bacteria Virgibacillus sp.SK39 into 50ml of CML medium, shake and culture at 37°C and 160rpm for 12h to obtain the host bacterial suspension;

[0022] The formula of the CML medium is: acid hydrolyzed casein 7.5g / L, yeast powder 10g / L, sodium citrate 3.0g / L, KCl 2.0g / L, MgSO 4 ·7H 2 O 20g / L, FeSO 4 ·7H 2 O 0.05g / L, NaCl 120g / L, pH 7.5;

[0023] (2) Mix the phage stock solution and the host bacterium suspension evenly, add 0.5% agar powder to obtain the upper layer medium containing the phage; wherein the upper layer medium contains 100 plaques and the plaques are dispersedly distributed;

[0024] (3) Add 1000 μL of VDS to the upper medium containing phage prepared in step (2), pipette repeatedly, and then vortex for 1 min to obtain a phage preservation solution;

[0025] The formula of the VDS is: NaCl 240g / L, MgCl 2 ·6H 2 O 30g / L, MgSO 4 ·7H ...

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Abstract

The invention belongs to the technical field of biochemistry, and particularly relates to a DMSO preservation method of halophilic bacterial phage. In the aspect of phage preservation, various preservation methods such as ultralow temperature cryopreservation (-80 DEG C), refrigerated preservation at 4 DEG C, preservation after freeze drying at 4 DEG C and DMSO preservation are known at present. However, the preservation effect of the phage of virulent phage Virgibacillus sp. SK39 separated and screened from the salt lake of the Yuncheng City is the best in which method, and no clear answer is available; and due to the fact that the structural forms of all the phage are different, the adaptive capacity of the phage to the environment is also different, and the preservation methods and temperatures are different. The preservation method has the advantages of being good in effect, simple in preservation condition and convenient to operate.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and in particular relates to a DMSO preservation method for halophilic bacteriophage. Background technique [0002] Bacteriophage is a kind of virus that infects microorganisms such as bacteria, actinomycetes, and spirochetes. It is small in size and simple in structure, and is widely distributed in nature. Wherever there are bacteria, there may be corresponding phages. Phages can infect their corresponding host bacteria. Based on this, phages can be divided into mild phages and virulent phages. The former adsorbs and infects the host After the cell, it combines with the host DNA and replicates with the DNA replication of the host cell. It does not replicate alone, does not proliferate and lyse, and the latter can proliferate, mature, and lyse in a short time after infecting sensitive host bacteria. After the phage infects the host bacteria, it can lyse and destroy the host bacteria through...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N1/20C12R1/01C12R1/92
CPCC12N7/00C12N1/20C12N2795/00051
Inventor 李新于慧瑛
Owner YUNCHENG UNIVERISTY
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