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Freeze-drying preservation method for bacteriophage

A storage method and phage technology, which is applied in the field of biotechnology and microbial products, can solve the problems of restricting the development of phage application, short storage time of phage, poor storage effect of phage, etc., and achieve electrolyte concentration maintenance, low storage environment requirements, and convenient application Effect

Inactive Publication Date: 2016-09-21
SUZHOU IRIVET BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, in the above-mentioned process, the phages need to be preserved. In the preservation scheme of the prior art, the preservation effect of the phages is poor, the preservation time of the phages is short, the survival rate is low, the phages are easily contaminated, and must be preserved under low temperature conditions. Seriously restricts the development of phage applications

Method used

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Examples

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Embodiment 1

[0027] A method for freeze-drying preservation of coliphage, which is completed under 4% humidity conditions, comprising the following steps:

[0028] 1) The Escherichia coli phage solution was left standing at 2°C for 4 hours, flocculent precipitates appeared, and centrifuged at 8000r / min for 40min;

[0029] 2) Add phage buffer solution to the above-mentioned precipitate, dissolve the precipitate, and obtain phage liquid; the phage buffer solution includes the following components in terms of weight components: 8 parts of Tris hydrochloride, sodium chloride 0.05 parts, 1 part of potassium chloride, 5 parts of magnesium sulfate, 1 part of dipotassium hydrogen phosphate, 8 parts of sodium dihydrogen phosphate, 0.05 parts of gelatin and 100 parts of deionized water, and use dilute hydrochloric acid and / or sodium hydroxide to adjust the pH to 7;

[0030] 3) The phage preservation and protection agent equal to the volume of the phage liquid is mixed evenly, and the amino acid of ...

Embodiment 2

[0035] A method for freeze-drying preservation of lactobacillus phage, which is completed under 6% humidity conditions, comprising the following steps:

[0036] 1) The Lactobacillus phage solution was left standing at 7°C for 8 hours, flocculent precipitates appeared, and centrifuged at 12000r / min for 20min;

[0037] 2) Add phage buffer solution to the above-mentioned precipitate, dissolve the precipitate, and obtain phage liquid; the phage buffer solution includes the following components in terms of weight components: 15 parts of Tris hydrochloride, sodium chloride 0.2 parts, potassium chloride 5 parts, magnesium sulfate 20 parts, dipotassium hydrogen phosphate 3 parts, sodium dihydrogen phosphate 12 parts, gelatin 0.1 part and purified water 100 parts, and use dilute hydrochloric acid and / or sodium hydroxide to adjust the pH to 7.5;

[0038]3) The phage preservation and protection agent with 2.5 times the volume of the phage liquid is mixed uniformly, and the amino acids o...

Embodiment 3

[0044] A method for freeze-drying preservation of Mycobacterium phage, which is completed under 5% humidity conditions, comprising the following steps:

[0045] 1) The Mycobacterium phage solution was left standing at 4°C for 6 hours, flocculent precipitates appeared, and centrifuged at 10000r / min for 30min;

[0046] 2) Add phage buffer solution to the above-mentioned precipitate, dissolve the precipitate, and obtain phage liquid; the phage buffer solution includes the following components in terms of weight components: 12 parts of Tris hydrochloride, sodium chloride 0.1 parts, potassium chloride 3 parts, magnesium sulfate 15 parts, dipotassium hydrogen phosphate 2 parts, sodium dihydrogen phosphate 10 parts, gelatin 0.08 parts and purified water 100 parts, and use dilute hydrochloric acid and / or sodium hydroxide to adjust the pH to 7.2;

[0047] 3) The phage preservation and protection agent with twice the volume of the phage liquid is mixed evenly, and the amino acid of the...

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Abstract

The invention provides a freeze-drying preservation method for bacteriophage. The method comprises the steps that bacteriophage liquid and a bacteriophage preservation protecting agent are mixed to be uniform to be prepared into freeze-dried bacteriophage powder, and the freeze-dried bacteriophage powder is preserved in ampoules in a sealing mode under the aseptic condition. The freeze-drying preservation method for the bacteriophage is reasonable and convenient to apply, and the bacteriophage can be preserved at room temperature or refrigerated, is low in requirement for the preservation environment and good in adaptability and can be preserved for a long term.

Description

technical field [0001] The invention belongs to the fields of biotechnology and microbial products, and in particular relates to a method for freeze-drying and preserving bacteriophages. Background technique [0002] Phage is a general term for a class of viruses that can infect microorganisms such as bacteria, actinomycetes, and spirochetes. No toxicity to animals and plants. With the development of molecular biology technology, phage has been widely used in many fields such as disease diagnosis and water treatment. Phages have also become important molecular and genetic research tools due to their ability to insert genes into host DNA. Using phages, many ingenious experiments can be designed. [0003] However, in the above-mentioned process, the phages need to be preserved, the preservation effect of the phages in the preservation scheme of the prior art is poor, the preservation time of the phages is short, the survival rate is low, the phages are easily contaminated, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12R1/92
CPCC12N1/04
Inventor 许维素宋增福陈彪
Owner SUZHOU IRIVET BIOTECH
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