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Shigella flexneri microphage SGF3 and application thereof

A technology of Shigella flexneri and Shigella, applied in the direction of phage, virus/phage, application, etc., can solve the problems of poor antibacterial effect, improve the breadth and effectiveness, and expand the host spectrum Effect

Inactive Publication Date: 2021-04-13
UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, considering the existence of the resistance mechanism between phages and host bacteria, when the pathogenic bacteria infection is more serious, the antibacterial effect achieved by direct use of one phage is not good, and it is undoubtedly important to choose multiple phages or the combined application of phages and antibiotics. practical significance

Method used

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  • Shigella flexneri microphage SGF3 and application thereof
  • Shigella flexneri microphage SGF3 and application thereof
  • Shigella flexneri microphage SGF3 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The separation and purification of embodiment 1 phage

[0021] The sewage sample used in the experiment of the present invention was collected in the west area of ​​Yanqi Lake Campus of the University of Chinese Academy of Sciences in Beijing in November 2019, and used as a water sample for the isolation of bacteriophage.

[0022] Centrifuge the sewage sample for 15min, take 25mL of sewage sterilized by filtration with a 0.22μm microporous membrane, add CaCl 2 Mother liquor to a final concentration of 1mmol / L, let it stand still, add 20ml NB liquid medium, then add 2.0ml of logarithmic phase host bacterial suspension, mix well in a 50ml centrifuge tube, let stand for 15min, shake overnight at 37°C ( 10h). The next day, add 2 mL of chloroform, centrifuge at 8,000 g for 30 min at 4°C, and the supernatant is the stock solution containing the phage of the host bacterium. Use the spot-test method to identify whether the stock solution contains phages. Take an appropriate a...

Embodiment 2

[0024] Morphological observation of embodiment 2 bacteriophage

[0025] Take 20 μL of the phage suspension and drop it on the copper grid, wait for its natural precipitation for 10 minutes, blot it dry from the side with dry filter paper, let it air for about 1 minute, add 1 drop of 1% uranyl acetate on the copper grid, stain for 2 minutes, and then use it carefully to dry Blot the excess dye from the side of the filter paper, let it dry naturally in the dark for 30 minutes, and observe it with a transmission electron microscope (JEM-1400).

[0026] The results of transmission electron microscopy are shown in figure 2 , the results showed that the phage SGF3 has a polyhedral structure with a diameter of 28±4nm. According to the eighth report of the International Organization for Taxonomy of Viruses (ICTV) virus classification, the phage SGF3 can be preliminarily classified as a microphage family.

Embodiment 3

[0027] Example 3 Determination of Phage SGF3 Host Spectrum

[0028] The 14 strains in our laboratory were used to determine the host spectrum of SGF3, including Shigella, Escherichia coli, Staphylococcus aureus and Salmonella. The infection of SGF3 to each strain was detected by the double-layer plate method, and each experiment was repeated 3 times, with two parallel samples each time. The infection status of phages to the strains is shown in Table 1.

[0029] Table 1

[0030]

[0031] Features: After the phage infects the bacterial strain, (+) has phage plaques, (-) has no phage plaques.

[0032] CDC is the Chinese Center for Disease Control and Prevention; CGMCC is the Chinese General Microorganism Culture Collection and Management Center.

[0033] It can be seen from Table 1 that SGF3 only specifically lysed the host strain 1.10599, and had no lytic effect on other strains.

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Abstract

The invention discloses a shigella flexneri microphage SGF3 and application thereof, and belongs to the technical field of biology. The preservation number of the shigella flexneri microphage SGF3 is CGMCC No.20294, and the shigella flexneri microphage SGF3 provides a selectable base material for phage treatment and prevention and control of pathogenic bacteria so as to enrich phage seed banks; through separation and identification, biological characteristic research and whole genome sequencing of the phage SGF3, functional genes and proteins of the phage SGF3 are further deeply excavated, one or more enzymes and the like with a cracking effect on the pathogenic bacteria are provided for the phage treatment and biological prevention and treatment, so that a host spectrum is expanded, and the universality and effectiveness of phage application are improved; and the phage SGF3 has a high cracking effect on shigella flexneri 1.10599, and a bacteriophage source is provided for industrial production of phage and prevention and treatment of shigella flexneri in sewage discharge and sewage reuse.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a microphage SGF3 of Shigella flexneri and its application. Background technique [0002] As a kind of Gram-negative bacteria, Shigella is also the most common pathogen of bacillary dysentery. It is mainly transmitted through food and drinking water, and has strong pathogenicity and lethality; HIV infection has become a public health issue of global concern. Among them, Shigella flexneri is the main pathogen of dysentery disease, and research on its prevention and control measures is imminent. [0003] As we all know, antibiotics are widely used to treat bacterial infections, but with the abuse of antibiotics, multi-drug-resistant and drug-resistant superbugs emerge in endlessly. Due to the rapid occurrence of superbug infections and the limited availability of antibiotics, antibiotic treatment has become a bottleneck, and researchers have begun to look for safer an...

Claims

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Application Information

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IPC IPC(8): C12N7/00C02F3/34A01N63/40A01P1/00C12R1/92
CPCC12N7/00C02F3/34A01N63/40C12N2795/14221C12N2795/14231
Inventor 刘新春卢晗
Owner UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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