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Nocardia cardia bacteriophage P3.2 and application thereof

A Nocardia and phage technology, applied in the fields of phage, virus/phage, biological water/sewage treatment, etc., can solve the problems of water deterioration, difficult to remove foam, etc., to improve the breadth and effectiveness, expand the host spectrum, Fast and efficient cracking effect

Pending Publication Date: 2022-05-06
UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, when Nocardia proliferates to a certain threshold in urban sewage treatment plants, the foam on the surface of the pool water will become stable and difficult to remove, resulting in deterioration of the effluent

Method used

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  • Nocardia cardia bacteriophage P3.2 and application thereof
  • Nocardia cardia bacteriophage P3.2 and application thereof
  • Nocardia cardia bacteriophage P3.2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The separation and purification of embodiment 1 phage

[0020] Preparation of PYCa liquid medium:

[0021] at 1L ddH 2 Add 1 g of glucose, 1 g of yeast extract, and 15 g of tryptone to O, mix well and then autoclave at 121°C for 20 min. After cooling, add 4.5 mL of calcium chloride solution with a concentration of 1 mol / L.

[0022] Preparation of PYCa solid medium:

[0023] at 1L ddH 2 Add 1g of glucose, 1g of yeast extract, 15g of tryptone, and 15g of agar powder to O, mix well and then autoclave at 121°C for 20min. Add 4.5mL calcium chloride solution with a concentration of 1mol / L in the uncoagulated state.

[0024] Preparation of PYCa semi-solid medium:

[0025] at 1L ddH 2 Add 1 g of glucose, 1 g of yeast extract, 15 g of tryptone, and 7.5 g of agar powder into O, mix well, and then sterilize under high pressure at 121 ° C for 20 min. Add 4.5mL calcium chloride solution with a concentration of 1mol / L in the uncoagulated state.

[0026] The sewage sample use...

Embodiment 2

[0029] Morphological observation of embodiment 2 bacteriophage

[0030] 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).

[0031] The results of transmission electron microscopy are shown in figure 2 , the results showed that bacteriophage P3.2 had a spherical head and a long tail. The length of the P3.2 tail was about 197.49nm, the diameter of the head was about 56.69nm, and the diameter of the tail was about 14.57nm. According to the eighth report on the virus classification of the International Organization for Taxonomy of Viruses (ICTV), this strain of ...

Embodiment 3

[0032] Example 3 Phage Genome Analysis and Identification

[0033] Phage whole genome sequencing and analysis: IlluminaNextSeq500 was used for PE 2×150 to sequence the DNA of the sample. Then, the optimized sequence was spliced ​​using spades v.3.11.1 splicing software, and the optimal assembly result was obtained. Use the biological software PHASTER to predict and analyze the open reading frame of the genome, and use NCBI Blastp to complete the preliminary annotation of functional genes, and use tRNAscan-SE (http: / / lowelab.ucsc.edu / / tRNAscan-SE / ) to predict tRNA online, Use CGView Server software (http: / / cgview.ca / ) to complete the drawing of the whole genome circle map (see image 3 ), a phylogenetic tree was constructed using MEGA X software based on the whole genome data (see Figure 4 ).

[0034]The genome size of phage P3.2 is 46380bp, and the G+C content is 67.1mol%. According to the results of Blast comparison, it is also a new phage, and the most homologous to it ...

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Abstract

The invention discloses a nocardia carnocardia bacteriophage P3.2 and application thereof, and belongs to the technical field of biology. The nocardia carnocardia phage P3.2 disclosed by the invention has the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.23093. The nocardia carnocardia phage P3.2 provides a selectable basic material for prevention and control of a sludge foaming event caused by nocardia carnocardia, and enriches a phage seed bank; through separation identification and whole genome sequencing of the bacteriophage P3.2, functional genes and proteins of the bacteriophage P3.2 are further deeply excavated, and one or more enzymes and the like with a splitting effect on nocardia carnocardia are provided for bacteriophage treatment and biological control, so that a host spectrum is expanded, and the application universality and effectiveness of the bacteriophage are improved; the bacteriophage P3.2 can rapidly and efficiently split nocardia carnocardia, and can be used for sludge foaming events caused by excessive propagation of nocardia carnocardia in a sewage treatment system.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a Nocardia carneurus phage P3.2 and its application. Background technique [0002] Nocardia is an aerobic, Gram-positive filamentous bacterium that is widely distributed in various natural environments and belongs to the Actinobacteriaceae. Mainly found in water, soil, dust and decaying vegetation. Nocardia is generally considered an opportunistic pathogen. To date, 55 species of Nocardia are known to infect humans. Some of the more prominent ones include Nocardia asteralis, Nocardia mallei, Nocardia guinea pig, and Nocardia brasiliensis, which are mostly caused by inhalation of pathogenic bacteria in the respiratory tract or infection by trauma, and are common in immunocompromised patients, such as Solid organ transplant (SOT) recipients. Nocardiosis may occasionally infect immunocompetent humans, especially those with chronic lung disease, bronchiectasis, diabet...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C02F3/34C12R1/92
CPCC12N7/00C02F3/34C12N2795/10321C12N2795/10331
Inventor 刘新春熊文斌卢晗
Owner UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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