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Rhodococcus pumilus bacteriophage P19 and application thereof

A rhodococcus and phage technology, applied in phage, virus/phage, applications, etc., can solve the problems of no wastewater treatment plant, no report on the isolation of Rhodococcus bacteriophage, lack of understanding of bacteria, etc., to expand the host spectrum, fast and efficient cracking , improve the effect of extensiveness and effectiveness

Pending Publication Date: 2022-03-25
UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been some solutions (such as adjusting sludge age, manual or mechanical defoaming, etc.), most of them are too specific and cannot completely cure foaming events. There is no general method to control the foaming events in wastewater treatment plants. bubbles, a situation that may reflect our current lack of understanding of the bacteria involved
However, there is no report on the isolation of Rhodococcus circulus phage

Method used

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  • Rhodococcus pumilus bacteriophage P19 and application thereof
  • Rhodococcus pumilus bacteriophage P19 and application thereof
  • Rhodococcus pumilus bacteriophage P19 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The separation and purification of embodiment 1 phage

[0021] Preparation of PYCa liquid medium:

[0022] 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.

[0023] Preparation of PYCa solid medium:

[0024] 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.

[0025] Preparation of PYCa semi-solid medium:

[0026] 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.

[0027] The sewage sample use...

Embodiment 2

[0030] Morphological observation of embodiment 2 bacteriophage

[0031] 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).

[0032] The results of transmission electron microscopy are shown in figure 2 , the results show that phage P19 has a spherical head and a short tail, and there seems to be a layer of envelope-like material outside the head; the specific dimensions are: the diameter of the head is about 55.44nm, the length of the tail is about 107.94nm, and the diameter of the tail is about 11.11nm. According to the eighth report on the virus classifica...

Embodiment 3

[0033] Example 3 Phage Genome Analysis and Identification

[0034] 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 ).

[0035]The genome size of phage P19 is 17065bp. According to the results of Blast comparison, it seems to be a highly novel phage, and the most homologous phage is Rhodococcus phage ...

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Abstract

The invention discloses a rhodococcus circulans phage P19 and application thereof, and belongs to the technical field of biology. The preservation number of the nocardia pekinensis bacteriophage P19 disclosed by the invention is CGMCC No.23087. The nocardia pekinensis bacteriophage P19 provides a selectable basic material for prevention and control of a sludge foaming event caused by rhodococcus pluvialis, and enriches a bacteriophage seed bank; through separation identification and whole genome sequencing of the bacteriophage P19, functional genes and proteins of the bacteriophage P19 are further deeply excavated, and one or more enzymes and the like with a splitting effect on rhodococcus circulans are provided for bacteriophage treatment and biological prevention and treatment, so that a host spectrum is expanded, and the application universality and effectiveness of the bacteriophage are improved; the bacteriophage P19 can rapidly and efficiently split rhodococcus circulans, and can be used for sludge foaming events caused by excessive propagation of rhodococcus circulans in a sewage treatment system.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a rhodococcus circulus phage P19 and its application. Background technique [0002] Activated sludge method is a commonly used aerobic biological treatment method for sewage treatment, which uses microbial activity and flocculation to remove biodegradable organic matter and suspended solids in water. Because the microorganisms aggregate into agglomerates and are in the shape of mud flowers, it is called the activated sludge method. [0003] Sludge foaming and sludge bulking are the main problems affecting its stable operation. Foamy sludge will reduce the amount of dissolved oxygen in the aeration tank, affect the quality of effluent water, and foam corruption will emit unpleasant gas. Sludge frothing is a flotation process that requires air bubbles, surfactants, and hydrophobic particles—usually bacterial cells. As the skeleton of activated sludge flocs, filamento...

Claims

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Application Information

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IPC IPC(8): C12N7/00A01N63/40A01P1/00C02F3/34C12R1/92
CPCC12N7/00A01N63/40C02F3/34C12N2795/10121C12N2795/10131C02F2303/20
Inventor 刘新春熊文斌卢晗
Owner UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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