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Method for efficiently screening low-temperature bacteriophages

A phage, low-temperature technology, applied in the field of efficient screening of low-temperature phage, can solve the problem of consuming a lot of time, and achieve the effects of improving efficiency, saving culture space, and saving costs

Inactive Publication Date: 2013-12-25
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The double-layer plate method needs to make two plates separately, and it takes a lot of time to prepare when it is used in large quantities

Method used

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  • Method for efficiently screening low-temperature bacteriophages
  • Method for efficiently screening low-temperature bacteriophages
  • Method for efficiently screening low-temperature bacteriophages

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 : The improved PYGV culture medium screens and isolates the method for low-temperature phage, and the specific contents are as follows:

[0033] (1) Isolation of low-temperature host bacteria: The soil or water samples collected from the Napahai Wetland in Yunnan Province were used to separate different low-temperature bacteria by the dilution plate method at 15°C using the solid-modified PYGV medium. Colony, as a possible host bacterium, pick a single colony with a sterile inoculation loop, inoculate it in the modified liquid PYGV medium, and cultivate it at 15°C and 150rpm for 16h until the concentration of the host bacterium reaches about 10 8 CFU / ml;

[0034] (2) Preparation of phage enrichment solution

[0035] The soil or water samples collected from the Napahai Wetland in Yunnan Province were added to the improved liquid PYGV medium (10g of soil samples were added to 100ml of medium, and 10ml of water samples were added to 100ml of medium), and pla...

Embodiment 2

[0046] Example 2 : The improved PYGV culture medium screens and isolates the method for low-temperature phage, and the specific contents are as follows:

[0047] (1) Isolation of low-temperature host bacteria: The soil samples or water samples collected from the Napahai Wetland in Yunnan Province were used to separate different low-temperature bacteria by the dilution plate method at 20°C using the solid-modified PYGV medium, and the single bacteria were obtained by streaking. Colony, as a possible host bacterium, pick a ring of colonies with a sterile inoculation loop, inoculate in liquid PYGV medium, cultivate at 10°C, 150rpm for 18h, until the concentration of the host bacterium reaches about 10 7 CFU / ml;

[0048] (2) Preparation of phage enrichment solution

[0049] The soil or water samples collected from the Napahai Wetland in Yunnan Province were added to the improved liquid PYGV medium (8g of soil samples were added to 100ml of medium, and 8ml of water samples wer...

Embodiment 3

[0059] Example 3 : The improved PYGV culture medium screens and isolates the method for low-temperature phage, and the specific contents are as follows:

[0060] (1) Isolation of low-temperature host bacteria: Soil or water samples collected from the Napahai Wetland in Yunnan Province were used to isolate different low-temperature bacteria at 28°C by using solid-modified PYGV medium as possible hosts. Bacteria, pick a ring of colonies with a sterile inoculation loop, inoculate in the modified liquid PYGV medium, and cultivate at 20°C, 150rpm for 12h, until the concentration of the host bacteria reaches about 10 8 CFU / ml;

[0061] (2) Preparation of phage enrichment solution

[0062] Add the soil or water samples collected from the Napahai Wetland in Yunnan Province to the liquid PYGV medium (10g of soil samples are added to 100ml of medium, and 10ml of water samples are added to 100ml of medium), and placed at 10°C Place in culture for 1-2 weeks, absorb the enrichment so...

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Abstract

The invention discloses an economical and practical method for efficiently screening low-temperature bacteriophages from an environment. The low-temperature bacteriophages can be screened by repeatedly utilizing a six-hole cell culture plate through a two-layer plating method. The component content of an improved PYGV substrate culture medium adopted by the method is one tenth of that of the conventional PYGV substrate culture medium, or the improved PYGV substrate culture medium is a one-tenth PYGV substrate culture medium which is short of 2X vitamins and / or contains 25% D-glucose. The content of the culture medium adopted by the method is only one thirtieth of that of the culture medium adopted by a common method. Thus, the method realizes cost conservation and is economical and efficient. In the same culture time and space, according to the method disclosed by the invention, the efficiency can be improved by 2.5 times or so. As a result, the method disclosed by the invention is suitable for the fields of separation of the low-temperature bacteriophages in the environments in laboratories and modern biological engineering.

Description

technical field [0001] The invention relates to an efficient and economical method for screening low-temperature phages, in particular to a method for substituting a six-hole cell plate for double-layer plate culture plates and utilizing an improved nutrient-poor medium for screening low-temperature phages. Background technique [0002] Phage is a general term for viruses that infect microorganisms such as bacteria, fungi, actinomycetes, or spirochetes. The distribution of phages is very wide. Wherever there are bacteria, there may be corresponding phages, and the total amount reaches 10. 31 Units have a profound impact on the evolution of organisms, especially other microorganisms. Due to the advantages of simple structure, small genome, and easy operation, phages are often used as models for the study of biological gene replication and expression regulation, and have made outstanding contributions to the development of modern biochemistry and molecular biology. [0003]...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12R1/92
Inventor 崔尹赡季秀玲魏云林林连兵张琦
Owner KUNMING UNIV OF SCI & TECH
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