Protective agent for freeze-dried preservation of genetic engineering mushroom strains and preserving method of genetic engineering mushroom strains

A technology of genetically engineered bacteria and protective agent, applied in the field of bacterial strain preservation and protective agent of genetically engineered bacteria, can solve the problems of low survival rate of bacterial strains, loss of recombinant genes, susceptibility to external adverse factors, etc. Effect

Active Publication Date: 2018-12-21
GUANGZHOU GENSOURCE BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, applying this method to the preservation of genetically engineered bacteria requires overcoming many difficulties. First, in genetically engineered bacteria, recombinant plasmids often show instability and are easily affected by external adverse factors, resulting in the loss of recombinant genes and the inability to express the purpose. The second is that the cell membrane or protein structure is easily destroyed by freezing during the freeze-drying process, and it is easy to be dried and dehydrated to death, resulting in a low survival rate of the bacteria after freeze-drying, which is not conducive to actual production and application

Method used

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  • Protective agent for freeze-dried preservation of genetic engineering mushroom strains and preserving method of genetic engineering mushroom strains
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  • Protective agent for freeze-dried preservation of genetic engineering mushroom strains and preserving method of genetic engineering mushroom strains

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Experimental program
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Effect test

Embodiment 1

[0013] (1) Construction of genetically engineered bacteria

[0014] Competent cells of Escherichia coli BL21(DE3) used in this protocol were purchased from Beijing Biomed Gene Technology Co., Ltd. A recombinant plasmid is introduced into the competent cells, and the recombinant plasmid contains a sequence encoding EGF protein. After verification, the target gene is introduced and successfully expressed EGF protein, indicating that the genetically engineered bacterium has been successfully constructed.

[0015] (2) Collect white fungus cultures and prepare strain fermentation products

[0016] Collect the remaining cultivars after harvesting the white fungus fruit bodies in the industry, dry the cultivars, dry them at 70°C until they are in an anhydrous state, and then pulverize until the particles can pass through a 180-mesh sieve, that is, the particle size is 80 microns to obtain the strain fermentation product.

[0017] (3) Preparation of protective agent

[0018] Weigh ...

Embodiment 2

[0041] (1) Construction of genetically engineered bacteria

[0042] The genetically engineered bacteria used in the scheme of Example 2 is the same as the genetically engineered bacteria used in Example 1, and will not be repeated.

[0043] (2) Fermentation of ash gray fungus, preparation of strain fermentation product

[0044]Weigh the fermentation medium of the ash fungus by weight percentage, and its composition and content are 45% of cottonseed hulls, 28% of bran, 13% of soybean powder, 1.6% of gypsum, and the balance is water. After the above-mentioned culture medium was sterilized by high temperature, the pure strain of C. cinerea was inoculated under aseptic conditions, and the inoculum amount was 0.5% of the weight of the culture medium. After the inoculation, place it at 26°C for constant temperature fermentation culture, end the culture on the 8th day, and collect the fermented product. The fermented product is dried, dried at 70° C. to an anhydrous state, and then...

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Abstract

The invention discloses a protective agent used in a freeze-dried preservation method of genetic engineering mushroom strains and a preserving method of genetic engineering mushroom strains. An ediblemushroom strain fermenting substance used in the technical scheme of the invention is used as the protective agent for freeze-dried preservation of the strains. Cultivation substances are fermented through edible mushroom strains such as cohabitant fungi, so that lignin and cellulose which are not liable to decompose at the normal state can be decomposed, organic compounds are fermented to decompose, and fermentation products and degradation substances contain various active micromolecules, such as micromolecular peptide, oligosaccharide and other low molecular weight substances of which thespecific chemical composition is still not found, which can maintain the integrity of cell membrane structure and protect the cell protein structure from being destructed. The implementation of the scheme proves that when an experimental group with the protective agent disclosed by the scheme of the invention is compared with a control group to which the protective agent disclosed by the scheme ofthe invention is not added, the technical effects of the experimental group scheme are obviously better, which proves that the strain fermentation substances have the advantages in the respect of maintaining the survival rate and the production properties of the strains.

Description

technical field [0001] The invention relates to the field of strain preservation, in particular to a protective agent used in the freeze-drying preservation method of genetically engineered strains and a strain preservation method of genetically engineered strains. Background technique [0002] In the field of biotechnology, the construction of genetically engineered bacteria is to introduce the target gene into the recipient bacteria and make it express, and secrete the protein expression that people need. After the construction of genetically engineered bacteria is completed, in order to maintain their excellent production performance and facilitate their use at any time in the future, the bacteria need to be properly preserved. At present, the vacuum freeze-drying method is one of the methods with the best preservation effect. Compared with other preservation methods, it can maintain the stability of the preserved strains to a great extent and prolong the preservation tim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12N1/21C12R1/19
CPCC12N1/04C12N1/20C07K14/485
Inventor 涂桂洪王曙聘潘忠林谭志康祝廷霞
Owner GUANGZHOU GENSOURCE BIO TECH CO LTD
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