CN116334108A - A Novel Anti-phage Component and Its Application - Google Patents

Abstract

The invention discloses a novel anti-phage element and its application. The novel anti-phage element has a nucleotide sequence as shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 or SEQ ID NO.6 . The novel anti-phage element found in the present invention can not only expand the library of phage-resistant elements and enhance the understanding of anti-phage element members, but also can be used to transform Escherichia coli to enhance resistance to different phages. Application value of fermentation industry.

Description

A Novel Anti-phage Component and Its Application

technical field

The invention belongs to the field of biology, and in particular relates to a novel anti-phage element and its application.

Background technique

Phages are viruses that infect bacteria and are the most abundant life form in the biosphere. Host bacteria, especially pathogenic and environmental bacteria, have evolved a variety of well-designed anti-phage elements during the game with phages. More classic anti-phage elements include modified restriction systems and CRISPR-Cas systems. Modification restriction systems Widely present in 75% of bacterial genomes, they are usually composed of DNA methyltransferases, restriction endonucleases and target recognition modules, which can quickly recognize and degrade specific phage-derived nucleotides . When the host bacterium is infected by phage, the CRISPR-Cas system will integrate its own CRISPR sequence into the phage genome. During the subsequent infection process, the caspase of the host bacterium can quickly detect and cut its specific sequence. in order to achieve anti-phage effect. In addition, anti-phage elements that have been paid more attention to in recent years include the reverse transcription subsystem, toxin-antitoxin system, etc., which can play an inhibitory role in various stages of phage infection of host bacteria, including phage adsorption, injection infection, Copying, releasing, etc. The rapid development of pan-genome sequencing technology has led to the discovery of more new anti-phage elements, which are usually clustered by two or more adjacent genes. In addition, related studies have found that genes encoding defense systems are frequently present in variable genomic islands or associated with mobile genetic elements. Recent studies have found that endogenous mobile genetic elements in marine Vibrio mediate the evolution of bacteria's resistance to phages, and the defense elements in a single bacterial genome can be as high as 6 to 12, indicating that the protection of phage defense elements is cumulative, and defense elements It accounts for more than 90% of the variable non-core genome, which explains why the biggest difference between many similar bacterial genomes lies in the composition and quantity of mobile genetic elements. Interestingly, these mobile genetic elements also travel fairly quickly among bacteria. Based on the widespread existence and rapid spread of anti-phage elements, it can be used for technical promotion and application. CRISPR-Cas9 gene editing technology has a huge impact on the medical industry. However, although different types of anti-phage elements have been gradually discovered, there are still few reports on its application in the fermentation industry and phage therapy.

Vigorous phage contamination in the fermentation industry is usually prevented by conventional methods, such as strain rotation, ventilation quality, and screening of resistant strains with phage receptor mutations, etc. result in impaired production. In addition, there are a variety of mild phages in the environment. After they infect the host, they integrate their genomes into the host genome, silence them, and enter the lytic cycle in a specific environment. The contamination of such mild phages is difficult to detect. But it will reduce production efficiency, and some may cause production failure. The discovery and transformation of new elements with high efficiency against phages can not only provide theoretical basis and resources for the development of anti-phage products based on phage-host interaction in the fermentation industry, but also can develop related medical agents targeting anti-phage elements. The aspect of medicinal bacteria has important practical significance.

Phages are always looking for host bacteria and trying to complete the infection process, which can be described as pervasive. In industrial fermentation production, engineering strains, especially Escherichia coli, are easily contaminated by phages. If the contaminated phages are not strong, they may even integrate into the host genome and coexist with the host in the form of mild phages, which is difficult to detect. They exist like time bombs, which may be activated at any time and enter the cracking cycle, killing the host and causing huge economic losses. In addition, phages are important disseminators of antibiotic resistance genes and host normal metabolism interference genes, and are one of the main factors restricting the production process. Host bacteria have evolved a variety of phage-resistant genetic elements in the process of fighting against phages, but most of them were identified through high-throughput screening methods, lacking systematic cognition. If we want to develop and utilize these anti-phage elements to assist the industrial production process of fermentation, we need to conduct detailed research on their function and mechanism of resisting phage infection.

Contents of the invention

The purpose of the present invention is to provide a new type of anti-phage element, which can enhance the effect of chassis cells against phage infection.

The present invention is to excavate a new type of anti-phage element, detect its anti-phage effect, analyze the breadth of its distribution, and transfer the element from different sources into Escherichia coli to detect the resistance to different E. The effect of enhancing the anti-bacteriophage infection of the chassis cells is achieved through the transformation of the chassis cells, thereby achieving the purpose of the present invention.

The novel anti-phage element of the present invention has a nucleotide sequence as shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 or SEQ ID NO.6 .

A second object of the present invention is to provide a chassis cell comprising the above-mentioned novel anti-phage element.

The third object of the present invention is to provide the application of the above-mentioned novel anti-phage element in improving the resistance of chassis cells to phage infection.

The fourth object of the present invention is to provide a method for improving the resistance of chassis cells to phage infection, which is to transfer the above-mentioned novel anti-phage elements into Escherichia coli to enhance the host's resistance to phages.

The chassis cells can be Escherichia coli, pathogenic bacteria Pseudomonas aeruginosa, Salmonella, Vibrio.

The phage may be phage PAP8, PAO-L5, QDWS, PAP-L5, Escherichia coli phage T1, T4, T5, T7, EEP, λ or M13.

The present invention focuses on a widely distributed brand-new anti-phage element, evaluates its anti-phage effect, and transforms the chassis cells by transferring it into Escherichia coli to enhance the host's resistance to phage.

Phage contamination often occurs in laboratories and fermentation industries, and some mild phages integrate their genomes into the host genome, making it difficult to be found and detected. The prevention and improvement of laboratory phage contamination problems mainly use methods such as cleaning related utensils and replacing bacterial strains. In the fermentation industry, phage pollution is usually prevented by means of cleaning the production environment and strain rotation, but there is no efficient and lasting solution. The invention creates and discovers a novel anti-phage genetic element, which has a good anti-phage effect. This new anti-phage element is widely distributed in different bacteria, such as pathogenic bacteria Pseudomonas aeruginosa, Salmonella, Vibrio, Escherichia coli, etc., indicating that they are very likely to have anti-phage effects on phages from different sources. Escherichia coli is the engineering bacteria that is most likely to be contaminated by phages in industrial fermentation. We introduced KKP anti-phage elements from different sources into Escherichia coli, which can enhance the effect of Escherichia coli against phage infection. Moreover, KKP elements from different sources have different resistance to different phages. If they are combined and designed, it is very likely to develop super-anti-phage engineering bacteria that are resistant to different E. coli phages, and to solve industrial fermentation from the source. Phage contamination offers a practical solution.

In summary, the novel anti-phage element found in the present invention can not only expand the library of phage resistance elements and enhance the understanding of anti-phage element members, but also can be used to transform Escherichia coli to enhance resistance to different phages. Strong application value of fermentation industry.

Description of drawings:

Figure 1 shows the genetic elements of KKP composed of three genes, pfkA, pfkB and PfpC.

Fig. 2 is the process of integrating KKP into the genome of Pseudomonas aeruginosa PAO1.

Figure 3 is the anti-phage effect of the three-component gene cluster in Pseudomonas aeruginosa, wherein -KKP is PAO1 that does not contain Pf6, and +KKP is the PAO1 that integrates the KKP three-component gene cluster.

Figure 4 shows that the KKP three-component gene cluster is widely distributed in prophages of many different bacteria.

Figure 5 shows that Escherichia coli carrying KKP from different sources has a strong ability to resist Escherichia coli phage infection.

Detailed ways:

The following examples are to further illustrate the present invention, rather than limit the present invention.

Example 1:

Salmonellaenterica subsp.enterica serovar Typhi str.CT18、Vibrio tasmaniensis10N.222.48.A2中的KKP三组分基因簇也进行了实验，它的KKP三组分基因簇序列如SEQ IDNO.2、SEQ ID NO.3、SEQ ID NO.4、SEQ ID NO.5、SEQ ID NO.6所示。1. Comparative analysis of the genomes of two filamentous prophages Pf4 and Pf6 co-existing in Pseudomonas aeruginosa MPAO1, and found that Pf6 carries two kinases (PfkA and PfkB) and one phosphorylase (PfpC) A three-component genetic element (Figure 1), referred to as KKP (kinase-kinase-phosphotase), its nucleotide sequence is shown in SEQ ID NO.1. The present invention is derived from Shewanella sp.W3-18-1, Escherichiacoli15EC039, Escherichia coli strain according to the same method

The KKP three-component gene cluster in Salmonellaenterica subsp.enterica serovar Typhi str.CT18, Vibrio tasmaniensis10N.222.48.A2 has also been tested, and its KKP three-component gene cluster sequences are as SEQ ID NO.2, SEQ ID NO.3, Shown in SEQ ID NO.4, SEQ ID NO.5, SEQ ID NO.6.

2. Since KKP cannot be knocked out from the MPAO1 genome, we integrated it into the PAO1 genome that does not contain Pf6 and is highly similar to the MPAO1 genome ( FIG. 2 ).

The specific operation is as follows:

A. Inoculate Pseudomonas aeruginosa MPAO1 from a -80°C refrigerator on an LB plate, culture overnight at a constant temperature of 37°C, pick MPAO1, inoculate it into LB liquid medium, and culture it at a constant temperature of 37°C with shaking until the OD ₆₀₀ is about 1.

B. Centrifuge 1ml of the bacterial solution at 12000rpm for 1min to collect the bacterial cells. Genomic DNA of bacteria was extracted using Tiangen Bacterial Genome Extraction Kit (Product No.: DP302-02).

C. Using primer pairs KKP-F1/R1, KKP-F3/R3 and KKP-F4/R4, using MPAO1 genomic DNA as a template. The primer pair KKP-F2/R2 was used, and the plasmid pEX18Gm was used as a template. Use Takara’s Primer Star reagent (product number: AL52850A) for PCR amplification (conditions: 95°C, 10min; 95°C, 30s, 60°C, 30s, 72°C, 90s, 35 cycles), and agarose gelation of the PCR product Gel electrophoresis, and then recover with the gel recovery kit of Omega Company (article number: D2500-02).

D. The vector pEx18Ap was double digested with EcoRI and HindIII, and then agarose gel electrophoresis and gel recovery (recovering PCR fragments as above).

E. The recovered vector and the above PCR fragments were ligated according to the instruction manual using the One Step Multi-Fragment Cloning Kit (Product No.: C113-01) of Novozyme.

F. Transform the ligated product into auxotrophic Escherichia coli WM3064 competent cells (0.3mM diaminoacrylic acid should be added to the medium when culturing), and use the pEX18Ap-F/R primer pair for the correct transformants Perform PCR verification and send to the company for sequencing. Thus, the WM3064 strain containing the recombinant plasmid was obtained.

G. Culture WM3064 strain (donor bacterium) containing the recombinant plasmid and Pseudomonas aeruginosa PAO1 (recipient bacterium) not carrying Pf6 until the absorbance at 600 nm is ~1.0. Mix 4 mL of donor bacteria with 1 mL of recipient bacteria, wash 3 times with LB medium, and centrifuge at 3000 rpm for 5 min each time. Suspend with 100 μL LB for the last time, drop on the LB solid plate containing 0.3 mM diaminoacrylic acid and 1.5% agar, and place the plate at 25° C. for static culture for 8 hours. Bacteria were serially diluted and spread on LB solid plates containing 30 μg/ml gentamycin and 1.5% agar, and the recipient strains successfully combined with transfer were selected. The successful conjugative transfer and the single-crossover strain containing the suicide plasmid were cultured at 37°C in salt-free LB medium (recipe: 1% peptone, 0.5% yeast extract, dissolved in distilled water), and then primed with primers PfkC-F and PfkA -R for PCR verification. Next, double-exchange screening was performed on the correct strains, and the salt-free LB solid medium containing 10% sucrose and 1.5% agar was used for screening. The obtained strains were screened for gentamicin resistance, and the primer pair conf-F/R was used for PCR verification for the strains that lost gentamicin resistance, and DNA sequencing was performed. Thus, the modified PAO1 integrated with the three-component gene cluster was obtained.

3. Use the phage of Pseudomonas aeruginosa preserved in the laboratory to carry out the infection experiment, and use the double-layer agar plate method to dilute the PAO1 cultured overnight at 37 ° C and the modified PAO1 integrated with the three-component gene cluster. , and then cultivated until the OD600 is about 1; at this time, mix 3ml of bacterial liquid with 10ml of R-top medium (temperature about 55°C) (recipe: 1% tryptone, 0.1% yeast powder, 1% NaCl, 0.8% agar) Spread it on LB plates and let it stand for 10 minutes in the ultra-clean bench for 4 kinds of Pseudomonas aeruginosa phage PAP8, PAO-L5, QDWS and PAP-L5 plate operation. Dilute the phage in a gradient of 10 ¹ to 10 ⁸ , pipette 5 μl each for spotting on a plate, and culture at a constant temperature of 37°C overnight, and detect phage resistance according to the number and strength of phage plaques. Results KKP showed resistance to phages PAP8, PAO-L5, QDWS and PAP-L5 (Fig. 3).

4. Using bioinformatics methods to predict, it was found that the three-component gene cluster is widely distributed in different prophages of various bacteria, including marine Shewanella W3-18-1 (P4 prophage), Salmonella enterica serovarTyphi str .CT18 (P4 prophage), Escherichia coli MPEC4969 (P2 prophage), Escherichia coli 15EC039 (P2 prophage), Vibrio 10N.222.48.A2 (P2 prophage) ( FIG. 4 ).

5. Synthesize the four three-component gene clusters in the above step 4 by gene synthesis, and clone them into the pHERD20T vector respectively. The enzyme cutting sites used in the vector are NcoI and HindIII, and Escherichia coli K12 strain MG1655 is used for transformation. Using the same double-layer agar plate method as in step 3, the coliphages T1, T4, T5, T7, EEP, λ and M13 stored in the laboratory were tested against phages. Only this time, 0.3% arabinose was used to induce the expression of the three-component gene cluster, and the induction time was 3 hours ( FIG. 5 ). It can be seen from FIG. 5 that the three-component gene cluster can improve the infection of the strain to phage.

Primers used in the present invention in table 1

ATGTTTCAAAGGCTATTGCAAAAACACCTTGCCAGAGGAATTCTTGGCAGAAAAATGTTATTCTATCGACAAAGGTTTCTATTGCCTTAGCTTCAGATCTAGGTCTGAAGAGAACTGAGAATCAAGACAGAACCGCTTTAATGAAATTTAGATCTTCAACAGCTTCGTATACTGTCATAGCCGTAGTTGATGGAATGGGTGGTATGAGAGATGGGGAGAAGTCTG CTGAAATAGCTATTTCAACTTTCTTGTGCTCTATTATGGAAAATGTTCATTTGGGTTCTGAACATGCAATAATGCAAGCCACGATGACTGCCAATAACGCTGTATTCGAATTTACAAATGGTAAAGGTGGAAGTACCTTATCCGCAATTTTATTAGCTAGCGACGGCACTCATATGACCGTTAATGTCGGGGACAGTCGAATTTATGCAAAGGAGTCTATCTTTGGC AAAGTAATTAGACTTACCGTTGATGATTCGTTAGCGGAAACCGTTGGAGGAAGCGGTACAGAATTATTGCAGTTCATAGGTATGGGGGAAGGAATTAGACCACACGTAGTTCCGCTGCCACTTGAAGCTAAGCAAGTATCTGACAACTGATGGTGTCCATTACATTGAACCAAACACATTGTCTGATATTATAAAACATGCAGAAAAAAATCACTCAGGTTG TAGAGCGGTTGATAGCAACTGCACGTTGGTGTGGAGGCCCAGATAACGCTACAGTTAGTGCTCTTGATTTAGAGCTATTAAACTTTGAGGAGTCTAAACTTTGAGGAGTCCCCTGGATGCCTCGATAGTGCAAATATCTGATCCACATAGCTCTACACAATTCATATTCCCCAATTTCAGTTGGAAAGTGAAGTATCTCCCAGAGACAAGTGTTCAAAATAATTCTATTAATACAG CTCAATCTGAGAAATCGGCTGCTCCTAGCAGAACCTCAACTTTGGTCGAAGAAAAAGAATCGATTACTGAAACAGTCAAAGTTGAAGCTAAAACCCCGCCTAAGAGAAAGAAACGTCAATCAAAGAAGGCTGTTGATCATCTAGATTCCGCAGATGAAGTACAAATTAAGATGACCATTTTTGATGAAGCAGGTTGCGAAAGTCATGAGGTTGAAGATGATGATTCCAAGTAGATATGAACTC TGTGGTAACAACGATACTGGTGGTATGGGTGACATTCTCTATTGCAAAGACAAACATTTACAACGCGATGTCATAAT TAAACTCTTAAAAGGTGATGCTGAACAACGAAGGTTAATTGACGAGCAAAAATCTCTTATTCAGCTCCGTTTCTAAAC ATGTAGTTCAATTGTACGACGTAGTTAATATTGATAATCAATCCGGTTTAGTATTAGAGTTTATTAAAGGCGAGGAC CTCAAAAATGGTCTTTATGAGTCGAATATGAAAGGACTCATAGAAGTATTGTGGCAAGTTGCGTGCGGTTTATCTGA CATACATAAGGCAGGAATTATTCATCGTGATATCAAACCAAACAATATTCGTCGAGATAACATCGGAGTTATCAAGG TCTTTGATTTTGGGCTTTCGAGAAAACTTGATAGCGCTAAAACACATAGTGTCATTGGTACTGTGGGGTATATGGCA CCTGAATTATGGAAATGTGGTGAAGTTGAGTTTACCACAGCTGTAGATGTTTATGCCTTCGGTATTACGGCAATGGC TCTTTCAAATGCTATTGTCCCAAGAGAACTTTTAGAGTTCCTCTCTCGTTGCGCTAACAATGGTTGGGTTAAAAATA GTTTACCAAGCTTAGATTCGGATATTGCAGAGATTCTAGAGCGATGCCTAGAATACAAACCAGAGGCTCGCCCCACA ATCGATCAGGTTGAAAAAATACTAAGAAAGCATCTCCTGAAAGACAAACACAAAGGTCTTATTGTGATGGGGAATGA AGTTAGAGAGCTTAATAATCAAAATAGACGTGCTAAGATTTCATCTATTCATGGTAATAATTTAAATGGTGAAATTA CCATTGAATATGATAGTTATGAATTTAAAGTTGCTGCTATCGGTGGTTCGGTAACAGTAAATAATGAAGTGATCAAAA GTCGGTTATATCCTGCCAGGTGCTTCCGTTATACTCTAGGTTCCGACTCTAATCGGCGTTTTGTTACTTTCGATAT ATCTAACCCAGAGGTGGTTTCATGATCACATCGAATACTCTGATCGGTGGCCGTTATATGGTTCACCAACACATTGGTGCTGGCGGTATGCAAGATGTTTATCTGGCGCTAGATCAATTCCTCGGTAATTATGTCGCACTTAAGACGCCTCAGCCTGGCCAGAAAACCAGACGATTCCAAGCTAGTGCTGTAATTGCAGGTAAGAGTAAATCACCACAATGTAGCCAAGACATTAGATTACTTTGAAGA AAATGGAAATGTTTACCTAATTGAGGAATTTGTTAAAGGTGAAACTCTTGAAGATAAGATTAAGCAGAGAAAATTCCTTGATCCTCATCTAGCCGCTAGAACTATACACCCTATTAGCAAAGGGGGTCAGAGCCTCACACATTCAAGGAGTAATACACAGAGATTTGAAGCCAAGTAATATTATGGTTGACTCTAGTACTGGTATCGAAGAGCTAAAGATTACCGATTTT GGTATTGCAACTTTTACCGACGAAGTATTTCAAGAAGAGGCCGACTCCGGCGATATTACTCGTTCCACTTCTGGGACAGTGAAAGGTGCTCTTCCATTTATGGCACCTGAAATGATGTTTCGTAAAAAAGGGGATAGTATTACTCCGGCCTTAGATATTTGGTCAATTGGGGCGATGATGTTCAAAATACTAACAGGTGAGTACCCATTTGGTGTTTT TCTCGATGCTGCTGTGAATGTCAAAAACAAGAAATAGGCTAGATTGGCCTTTTTATGACTTCTAACGCACAGTTTTTCCCCATTATGCAGGGAACTTCAGAAGATAATAGATAGCTGTTTGGAGTATGAACCAACCAAACGCCCTACGGCTGATGCTCTTGTGAAAATGTGCCAAAATTTGTGCTATCAAACTTCTGAACGTTTTGAAGCGACGGTTACGAGGAT GATTCAAAAACGGATATAGTGGCTTTGCTTCAAACCCTCAACATAGCGTATTTTTAGTATCCATAGCATCTACGGAGCTTCTAGGGTTAATAGTGGAAGTAAAATTACGTACTCAAAGTTTCCGGGCACCCCTAATTTTAGAGCTCACCCAGTCATTATCTTAAATTAA

GTGAGCCGTGATTCTTACGAAATCCTTCATGAGCATATTCACGGATGGCTACATCGAAAAAATATAGCATCCTCAGTGCGTCGTGTCTCAACCTTACCAGTGGCTATAGCTACTGATATTGGGTTGGTACGGAAGGAAAACCAAGATAGGGTTGCTATATTGAAATTCCGCCCAAGTAGCAAAGCTAAAGATATCGTTGTTGTTGCGTTAGCCGATGGTATGGGGGGGATGGAGGGGGTGCCAATGCAGCATCTTTAACTTTATCTACATTTTTACTGAAATAATAAGAAATTCTCATTTACCAATACGTTCTTGTCTTGAGAAAGCTGTGCTACAAGC GAATGATTCTGTTTTGAATGTATATAAGGGTAATGGAGGGGCGACATTATCAGCGATAGCTTTAGAAGATGATGATAATATCCGCAGTAAATGTAGGTGATAGCCGTATTTATTATGTCTCGCATGAAGAAACTACTCAGTTGAGCGAGGACGATACTTTAGTTGCCTTAGCCAAAAGTATAATAATCATCTAAATATGGATCCACAGGATATTGATTTACG CTTTGGTGGCGAATTAGTACAATTTATAGGCATAGATGGCACATTGGAAATACACTTTTCATCATATCCAAGCCTTGGAGAGTGGCGTAATTATTTTAAGTTCTGATGGTGCGCATTCTATCGGTAAGGATAATTTAAGAAAGTTATATGTGCACTCGGCGAATCTAGGTGTTTTATTCTAGGCGGGTTATTGATCTTGCTAGTTGGTTTGGTGGTTTCGACAAT GCAAGTATAGCAGTTATAGACCTTTTTAATACTCTAAAAGAGTTAGATGTTTCATCTGGAGATGTAATAAATCTCTGGGATCCATTTGGTGAATTAAAAGTCATTAGCGTGCCGAATAAGTCATCTTCATTAGAGCCTTTAAATACAGTTGAGCTTGAAAAAAATAATAAAGGCACTTCTGGTGTGAAAAGTGTCAAAAAAAAATACTGATTCAGTCGTTAGCGATA TAAACAAAGCAACAACAAAAAGAAAGGCAAGAACAAAAAATAAGAATAAAATCTCTTCAGGAATTAGATGAGAAAAAAAATGGGTTAAATAAAAATAATAGACTTGATTGTATTTCACAACTTGACATGTCATTCCTTGAAAGGAATTCAATAAAAGGAGATGGTGATGATGAGTGATTTTCTCCCCAGAAAGATATCAAGTGGT TGGGGATCCTGATTTAGGGGGATTCGGTAGTGTAATCAAATGCCGCGATTCTCATCTTGAGAGATTTGTTGCAATAA AGACTATAAATGATCCATCAGATACAGAGCGAATGAAAGACGAGTTGGCTGCTCTAATGACACTACGTTCAAAACAT GTTGTTGAACTGTTTGATGTGATTAATTATGCTGAAGGCAATCTTGCAATTGTTGAAGAGTTTATCGATGGTCCATC GTTGAATGAAGTTAATAATAAAATTACTACAGTAGGGGAGCTTATTAAGATTTTGTGGCAAATAGCATCAGGTATTT GCGAGATACATGAACATGATATCATTCATCGTGATATAAAGCCTGGGAATATGAAGATTGATAAAGAAGGGCTTGTA AAAATATATGATTTTGGCTTGTCAAGAAAAATAGATAATGCAAAAACAATTGGGTTTAAAGGTACCCCAATTTTTGC AGCTCCTGAGTTGTATTTGCAGAACGTAGACTTTACTAAAGCAATTGATACATATGCCTTTGCTGTTACAGCAATGT GCTTAGCTAAAACCCCTGTCCCAGATGAATTGACCCGTTACCCTAAAGATTCTGACATCTAATCCATTTGATTTGTCG GTAATAAAAATTACCAAGTATTGTAAAGGAATTGTTTTTCAAATGTCTTGATGCAAATCCTCAAAGCTAGGCCCCCTAT GAAAGATGTTTGCGATGTTTTGAAAAAAAATATTATTGCACAACTCTCATCGAGCATTGCTTATATCTGATAATAAAA AACCAGTAGTGCTCTCAGGCTACACACAAGACGGAGTCTTATAACAATCCAGGGGTGGGTAGTGTGGAAATTACTTAC TCTGGTTCCGAGTTTTATATTTCAGATATCAGGGGATGTCTATGTTAATAACATTAGGGCTAAAAAACGAAATTT ATTGCCTAGCTCATGCGTGATAATACTTGGCCCTGCCGGAAGAACAACTACAAAACGTATATTTATCACATTTGATC TTTCTCATCCGGAGGTTGTGTTATGATTGAGTTGGTTCCTGGGACTAATAATAAATCGTTATACTATTATCAGCGAAATTGGTGAGGGGGGATGCAAAAGGTTTACCTTGCGAATGATAAGATATTAATAGGCAAGTTGCTCTTAAGACCCCTAAAAATAAGTCTGCTGAAAACGATTCCATAGAAGTGCTATTTTAGCATCTAGGGTCAACCATCCTAACGTCGCTAAAACATTAGATTA TTTTGCCGAAGATGGACGTGAATTTTTAACGGAGGAATTTATCGATGGAGTAGATCTGGATAAAGCATTGTTGAGTAGCTATACAAGTGTTGATCCTACTTGACTGCAAAGATATTTCATAACTTAGCGAAGGCTCTTTCGGCTTCCCATCATGTGGATGTAATACATCGAGACCTCAAACCTTCTAACATAATGGTTATTGGAGGAGTTAGTGCTACAGGT GTTAAAATCACCGATTTTGGAATTTCAAAAAATGGCCGGTGATGAAATTGATGAGGCCGCAAAGAATGGGCAAGGATCGATTACTTCATCTCAAACAGCTATGGGGGCATTGCCATATATGGCCCCGGAAATTATACAAAGTCAGGGGCAAGTTTCAAAACCATCTGATGTCTGGGCATTAGGTGCGATGATGTTCAGAATCCTCACGGGAGA GTATCCTTTTGGATTAGGGTATATGGCTATTCCGAACATCTTATCTGGAAAGCATACTCAATATCCTGATTTTATTAAGTCAAATAAGCAGTTTGCTCCGCTGGCAAATGAAATTATAGATAATAATTGAAAAATGTTTAAATCTAGACCCTTCTAAACGCCCCACTGCAGATGAGCTCGTGTCATTATGTGGTCAATTATGCTATCCGGTTTGTAATAGAGAAGAAGGAG TAATAGGTGATACTAGACTAGCTTATGGTTTCATTCGTATACCAAACCAACCACAAGTATTTTTTCATTACGATAGTGTGTATGGTAGTAAACCAGTGAGTAATGATAAGGTGATTTTTTCAAAGTTCTTGGGAGGGGGCCATGACCGGGCTCATCCAGTTATCAAGGCTAAGTAG

GTGAGCCGTGATTCTTACGAAATCCTTCATGAGCATATTCACGGATGGCTACATCGAAAAAATATAGCATCCTCAGTGCGTCGTGTCTCAACCTTACCAGTGGCTATAGCTACTGACATTGGGTTGGTACGGAAGGAAAACCAAGATAGGGTTGCTATATTGAAATTCCGCCCAAAGTAAGCAAAGCTAAAGATATCGTTGTTGTTGCGTTAGCCGATGGTAT GGGGGGGGATGGAGGGGGGTGCCAATGCAGCATCTTTAACTTTATCTACATTTTTACTGAAATAATAAAGAAATTCTCATTTACCAATACGTTCTTGTCTTGAGAAAGCTGTGCTACAAGCGAATGATTCTGTTTTGAATGTATATAAGGGTAATGGAGGGGCGACATTATCAGCGATAGCTTTAGAAGATGATGATAATATTACCCGCAGTAAATGTAG GTGATAGCCGTATTTATTATGTCTCGCATGAAGAAACTACTCAGTTGAGCGAGGACGATACTTTAGTTGCCTTAGCCAAAAAGTATAATAATCATCTAAATATGGATCCACAGGATATTGATTTACGCTTTGGTGGTGAATTAGTACAATTTATAGGCATAGATGGCACATTGGAAATACACTTTTCATCATATCCAAGCCTTGGAGAGTGGCGTAATTATTTTAAG TTCTGATGGTGCGCATTCTATCGGTAAGGATAATTTAAGAAAGTTATATGTGCACTCGGCGAATCTAGGTGTTTTATTCTAGGCGGGTTATTGATCTTGCTAGTTGGTTTGGTGGTTTCGACAATGCAAGTATAGCAGTTATAGACCTTTTTAATACTCTAAAAGAGTTAGATGTTTCATCTGGAGATGTAATAAATCCTGGGATCCATTTGGTGAATTAAAAG TCATTAGCGTGCCGAATAAGTCATCTTCATTAGAGCCTTTAAATACAGTTGAGCTTGAAAAAAATAATAAAGGCACTTCTGGTGTTAGAAAAGTGTCAAACAAAAAAATACTGATTCAGTCGTTAGCGATATAAACAAAGCAACAACAAAAAGAAAGGCAAGAACAAAAAATAAGAATAAATCTCTTCAGGAATTAGATGAGAAAAAAAATGGGGTTAAATAAAAATAATAGACTT GATTGTATTTCACAGCTTGACATGTCATTCCTTGAAAGGAATTCAATAAAAGGAGATGGTGATGATGAGTGATTTTCTCCCCAGAAAGATATCAAGTGGTTGGGGATCCTGATTTAGGGGGATTTGGTAGTGTAATCAAATGCCGCGATTCCATCTTGAGAGATTTGTTGCAATAAAGACTATAAATGATCCATCAGATACAGAGCGAATGAAAGACGAGTTGGCTGCTCTAATGACACTACGTTCAAAACATGTTGTTGAACTGTTTGATGTGATTAATTATGCTGAAGGCAATCTTGCAATTGTTGAAGAGTTTATCGATGGTCCATCGTTGAATGAAGTTAATA ATAAAATTACTACAGTAGGGGAGCTTATTAAGATTTTGTGGCAAATAGCATCAGGTATTTGCGAGATACATGAACATGATATCATTCATCGTGATATAAAGCCTGGGAATATGAAGATTGATAAAGAAGGGCTTGTAAAAATATATGATTTTGGCTTGTCAAGAAAAATAGATAATGCAAAAACAATTGGGTTTAAAGGTACCCCAAATTTTTGCAGCTCCTGAGTTGTATTTGCAGAACGTAGACTTTACTAAAGCAATTGATACATATGCCTTTGCTGTTACAGCAATGTGCTTAGCTAAAACCCCTGTCCCAGATGAATTGACCCGTTACCCTAAGATTCTGACATCTAATCCATTTGATTTGTCGGTAATAAAATTA CCAAGTATTGTAAAGGAATTGTTTTTCAAATGTCTTGATGCAAATCCTCCAAGCTAGGCCCCCTATGAAAGATGTTTGCGATGTTTTGAAAAAAATATTTATTGCACAACTCTCATCGAGCATTGCTTATATCTGATAAAAAACCAGTAGTGCTCTCAGCTACACCAAGACGGAGTCTTATAACAATCCAGGGGTGGGTAGTGTGGAAATTACTTACTCTG GTTCCGAGTTTTATATTTCAGATATATCAGGGGATGTCTATGTTAATAATATTAGGGCTAAAAAACGAAATTTATTGCCTAGCTCATGCGTGATAATACTTGGCCCTGCCGGAAGAACAACTACAAAACGTATATTTATCACATTTGATCTTTCTCATCCGGAGGTTGTGTTATGATTGAGTTGGTTCCTGGGACTAATATAAATCGTTATACTATTATCAGCGAAATTGGTGAGGGGGGGATGCAAAAGGTTTACCTTGCGAATGATAAGATATTAAATAGGCAAGTTGCTCTTAAGACCCCTAAAAATAAGTCTGCTGAAAAACGATTCCATAGAAGTGCTATTTTAGCATCTAGGGTCAACCATCCTAACGTCGCTAAAACATTAGATTATTTTGCCGAAGATGGACGTGAATTTTTAACGGAGGAATTTATCGATGGAGTAGATCTGGATAAAGCATTGT TGAGTAGCTATACAAGTGTTGATCCTTACTTGACTGCAAAGATATTTCATAACTTAGCGAAGGCTCTTTCGGCTTCCCATCATGTGGATGTAATACATCGAGACCTCAAACCTTCTAACATAATGGTTATTGGAGGAGTTAGTGCTACAGGTGTTAAAATCACCGATTTTGGAATTTCAAAAATGGCCGGTGATGAAATTGATGAGGCCGCAAAGAATGGGCAAGGATCGATTACTTCATCTCAAACAGCTATGGGGGCATTGCCATATGGCCCCGGAAATTATACAAAGTCAGGGGCAAGTTTCAAAACCATCTGATGTCTGGGCATTAGGTGCGATGATGTTCAGAATCCTCACGGGAGAGTATCCTTTTGGATTAGG GTATATGGCTATTCCGAACATCTTATCTGGAAAGCATACTCAATATCCTGATTTTATTAAGTCAAATAAGCAGTTTGCTCCGCTGGCAAATGAAATTATAGATAATTGAAAAATGTTTAAATCTAGACCCTTCTAAACGCCCCACTGCAGATGAGCTCGTGTCATTATGTGGTCAATTATGCTATCCGGTTTGTAATAGAGAAGAAGGAGTAATAGGTGATACTAGACTA GCTTATGGTTTCATTCGTATACCAAACCAACCACAAGTATTTTTTCATTACGATAGTGTGTATGGTAGTAAACCAGTGAGTAATGATAAGGTGATTTTTTCAAAGTTCTTGGGAGGGGGCCATGACCGGGCTCATCCAGTTATCAAGGCTAAGTAG

ATGTTTACAGAACGACTTGCTCGCTGGTTAGCTCGTTCTTCGGCCAAAAGCGGCATTAACCGGCCAGAAGACCTCAACGCTGTCCTTAGCACGGATATAGGACTGGTTAGAGCTGAGAATCAGGATCTAATAGCCGCGATTAGAGTTAACACTCCGTCAAACGTTGGCAATCCTTTTTTTGCAATGGCGTTATTAGATGGCATGGGTGGAATGCA AGATGGAAAGCAATGCGCAACAATTGCTTTATCAACTTTATTCTATTCTTTGATTAAGTTTAGAAGTGATCCTCCCGAGTCTCGGTTATTAAAAGCGACTTTAGAAGCAAACTCCGTTGTATATGACTATGCAAAAGGACATGGCGGTTCAACATTTCCGCTGTAATTATTGAAAATGGGTCTGCTCCTGTAATTGTCAACGTTGGCGACAGTCGAATATAGCT TTTCTCTGGACTGTGGACTTACAGCAATTAGCAGTGATGACTTCTCTAGAAGCATTGGGTGGCAGAGGGCGCGGATTACTCCAGTTTATAGGAATGGGGGAATCTATAAAGCCTCATATCAATATCTTAGATAAAAATCATAAAAAATAATATTGACATCCGATGGAACTCATTTCATTTCTCATTCAGCATTCGAAGAGTTATTAAGTCATTCATCTGATTTTTCT ACATCAGCGCAGAGAATAGCTCAATATGTTCAATGGTGCGGTGCGAAAGATAACGCTTCATTTGGAATTAATTAATTGCAATGATATAGAAAACAGCCTCAACTCCCATAAAGATATTGGTGTAGAGTTATGGGACCCTCATGGGAATCTACATATCATGTGGATGAAAAAATTATCCTGCAGCGCAGAATTACTTCTCTCAAATATAGTGGATGATCAAGATAAA GAACCTTCACCTATTTATAGACGATGATGGTTTTGAAAATAAAAAAACACTAAACAATCCTTCAACAAAAAATCTAGAATTAGATTCTGAAACACCACAAAGAGAGCTATTCTCAAACGAATCCCCAGAAAAATCTCAAGATCCATCCATTACAAGCAAAGCAATCAAACAAAGAAAAACAAAGATAAAAAGAAAGCTATTGAAAAGATCAAAAAAGACCAATCTGTAATGATAAATAT CAAGGATGAGGAAA ACAAAAATGAAGATTAATCACGTCCTACCAGAAAGATATTCATTAAAGAGCACTGAACTCGGTGGTGGCATGGGGGACATTTTAATATGTAAGGATAATCATTTAGATAGAGATGTAATTGTAAAGTTGTTAAAGGATGGAGAAGAAGAGAGACGTTTACTAGATGAACAGAAAGCGCTACTCAAACTTCGTTTCTAAACATGTAGTACAACTTTATGATTTAATTGACATAACAGTCTCCGAAAAAACTAAAAAAAGGATTAGTTCTGGAGTATATTAACGGAGTGGATTTAAATTA TAACCCGTCAGAAAGTCACCCCCGAAAAACTAAAAGAAATTATGGCAGATAGCATGCGGGTTAAGTGACATCCACTCTGCTAAAGTAATTC ACAGAGATATAAAGCCCAATAATATTAGAGTAGACGAGAATAAAATTGTTAAAATATTAGATTTTGGTTTAGCAAGGACCTCAGGCACAGAAGCATTCACTCATTCTGTTATTGGAACCTTAGGATATGGCTCCTGAACTATGGAAGAGAAAAACATTAGTTTCGATCAAAAAATTGATGTTTATGCATATGGTGTCCTCGTTTTAGATTTATTCGGCATAGAAAAACCAGATGAATTATACGAACTCCCCCGGCCGCGATAACCAATATACCTGAATTAGGAAAGATA CTTCCAAAGGACTTAGCCAGAACTTTTCATTAGTTGCTTAAGCCATGACAAATATGCTCGACCGGCAATGTCTTCGGTTAGAGATCAAATAGCTAAATACCTATTAAAGATAGACACCGTGCCCTCTTCGTCCTGAATGGAAAGAAATATGAAATAAATGCTAAAAAATAAAAGTGTTACGATCACTTGGGGTACTAGTGGAAGTATGGAAATAGTATATGATG GTTTCGACTTTAAAGTAGGTAATTTTTTCAGGAAGTGCGACAATTAATAACCAACAAGTGATAACAAATAAAGTATTTCCTAGCTGCAGTGTAATTACTCTTATAAATGAAAAATCTAGAAGCTTTGTCACCTTTGATATATCTAGACCGGAGGTAATATCATGATTGAAGTCGGAAGAATCATTGCAGAACGCTATAAGATTTTATCTTATGTTGGTAAAGGTGGGATGCAAGACGTATATAAAGTTTTAGATCTAAAGCTGGATTTAGATTTAGCTTTAAAGACTCCACTTCCTGGTTTGGAAAGCAAAAGATTTCTTAAAAGTGCCAAAATCGCTGCAAAAATAAAACCATCACAATATAGCAAAAACCTTTGATTATGTTGAAGATAATGGCAATATATTTTTAGCGGAAGAATTTGTTGAAGGTGAAAACCTTGAGGAAAAATTGGCGCCACTTT GATTTTTTAGACCCACATTATGGCGCTTGTATACTACATAACCTTGCTAAAGGAATAATGGCATCTCATAAAGCAGATGTTATTCATAGAGATTTAAAACCGAGTAATGTAATGGTGTCTGGCGGAGTACAAAATTTCGAATCTAAAAATTACAGATTTTGGAATAGCTACGCTAACACAAGAACTTTTTGATGAAGCTGCTGCCAGTGGTGACCTAACAAGATCGACCTCTGGTACGATAAAAGGTGCTTTACCCTTCATGTCTCCTGAATTAATGTTTGGTAAAAAGGGTAAACCTATAGAAGCATCAACTGATATTTGGCCATTAGGTGCAATGATGTTTAAATTATTAACAGGAGACTATCCATTTGGCGTTTATTTAGATGCT GCTGTTAATGTTAAAAACAAAAATAGAATGGAATGGCCAACCTTTATGACTGCCAATCCACAATATCAAAGTTTTATCACAAGATCTACAAAAAATTGTAGATAAGTGCTTAAGTTATGACTCGGACAAGAGGCCTACAGCTGAAACGCTTGTTAAGGCATGTGAAACCCTTTGTTATTTATCCGAAGAACGTCATGTCGGTCGCGTCAATAATCTTATTCAAGGAGGGA TAAGTGGATTTATTGATGGGACACCTTCTAACTCCTTTTTTTAGTATGGAAAGTGTATACGGTTCGAGGTACCCAAAACACCTCAACAAGAAATACTGTTTGTTATTTCCACTTTTGATGGACACCCATGGCCTAGAGCACACCCTGTAATTCTTTGGAAAGATTGA

ATGCAAGATATATTAACAAAAAGGCTAAATAGACCTGTAAATGGGAACCGGTCATCCGTTGTACATGAAGTCGGAGCGACGTTAGCGACAACTGTCGGGCTTATCAGAACTGAAAATGAAGATCGGGCAATTTCTGCTCGTTTCTATAGTTCAAAAAATGGAGCGCTACATAACTTTTATATTCTTAGTGATGGTATGGGAGGTATGGTTAATG GGGGGCTTGCCGCAACTCACACTGTCTCCAATGTTTTAAGCCTATAATACCCTCTTATCGAGTCAGGCATTGAAATAGATCAAGCGATACAACAGTCTGTGTTTTCTGCTCATCAGATCGTTTTCCGAATCAACAAACGGTAAAGGTGGAGCAACTTTATCAGCTATAGTCAGCCATGAACCAGGTGAATTCTTTACCGTCAATGTTGGTGATAGTCGAATTTATCAATGC ACAACAAGCAATTCTATTTTTCAAATCACTGAAGATGACGATGTAAAAGCTTCTCGAAAAACTCAACGGAATCGAACTAAACAGCCTAATAACGAAGAGAAATGGTCTAACAAGTATAGGCATGGAGGGAGAATTAGAGGTCACTGTTGAGTCATGCTCTGCACTCTGCGATTTGTTAGTCTATTCTGATGGTATAAGCCAATTGGAGAAGCTAACTTAGT TGGCCTATATGAAAATAAGACAACCGATAGTGAATTCGTTCAACGTTGTATACATTTGTCCAATTGGCTTGGCGGTCATGATAACGCAACAGCAATCTATGCTTCTTTGGCAAACCTAACATATCACCCAGAAACTACCGAAGTCACCCATAACTGCCTTGAAGTTTGGGATTGCCATGGTTACATTCATTATTCCCTTAGCACAACTACCTTCGAAAGGTAGACCC AAAAGAAAAAACGAAAAAGTACGAAAAAAAACAGCTGCGAAAAAAAATAATAAAAAGTGATAAAGAAGACTCTCGCAACGATTATAACCTAGAGATAAATCAAAAATCATTATTCTCATCTGACGATGTCGGAGAGATTTCGCCTGATAATAACAGTGTATTTGATGAAGGTGCAGGACTTGACCTAGCCAAAATCGAAGATAATAAAAATAATCTAGATAAATAGAATTAAACGT TGGTGACAATTATGGCTAGTACAAGATATGAATACCTAAAACACATAGACGATGGCGGTTATGGTAGTGTCCTCTTTTTATAGAGATATATTCCTTGATCGTGAAGTCGCAATAAAAACAATCCCCATTAGTAAGAAGAACAATACGATTGAGGAAGTAAACCTACTCAAATCAATTGCGTCTAAGCATGTTGTTGGATTGTATGACGTTATTCAAACCCAAACTAACATTGAAATATATCAAGAATACTTAGATGGGGAAGATCTAGCTAGCAAAGTAGGTCAGTGTCATGG CCCAGAATTTTTGAGCCTTGCCTATCAGCTAGCATCTGGACTACGTGATATACATTCATCAGGAATATGTCACCGAGATATAAAGTTGGATAATGCGAAATTTGACAAACAAGGTGTACTTAAAATTTTTGATTTTGGTGTATCTCGTATCGGTGATCCGCACCAAACCGTAAATGGTCATGGTACATTAGAATACCTAGCTCCTGAAGCCTTTGGATTATACACGCAAGACTCCGTTGTACTTTCATTTGCGGTCGATATCTATGCACTTGGAGTGACCTTACACAAACTTGCATTTTCGGGTATATGTAAATCAATAAAACGCTTAATCCGCAACCAGAAAGCCCTCGATTTGCTGAGC TTGGCTTTTGTTCAAAGCTAACATCACTCTTAAATAAATGTGTTTCAGAAAATAAAACGGAGAGACCCTCAGCAAACAGTTTAGTTGCTGAATTACAAAGAGAGCTCCTACGAAATAAACATACTGGTTTGTTTGTTGCACCGAAAGGCTCCCATAATATTGACCAAGCTCATCCAAAAACAAGAATTAAAATTTCTGATGACTTGTCAATTATTATCAATTACAACGGATAC GACTTTTTATGTTACTAAAGTTGAAGGACATGTTATATAAATAATGAAATTGTAGATGTGGGTAAGGTGCTTTATGGAGCATGCGTCCTGACATTTGTTCGAGATGCCAATAACCGTTTCTTTGTTTCGTTTTTCTTCATCTCATCCGGAGATAGTATTATGAGTCATATTCACAAACCTAACGACATCATTGCAGATCGATACGTGATCGAAAATTATATCGACGAAGGTGGTATGCAACAAGTCTACTCAGCGATAGACAAAAACATTGGACGAAAAGTCGCTTTAAAGACACCAAAAAATGATTCGGCTAGCCTGAGATTTAAAAGTACCGCAATTTGTAGTGCTCGTGTTATTCACCCTAACGTGGCGAAAACTTCTAGATTACTTTGGCTTTGATAAACGCGAATACTTAATTGAAGAACTTATTGACGGAAAAGATTTAAACACTGTATTCAGAAAATAA TTTCTCATATTTAGATCCATGTCTCGTTGCCTTCATAGGACATCATCTATCAAAAGCAGTAGCTGCATCTCATAGCGCAGACGTCGTCCAT AGGGATCTCAAACCAAGTAACATTATGATTGTAGGGGGCGAGAAGTTTAGAGATATTAAAGTCACTGACTTTGGTATTGCAAAACTTGTTGATGATGAAATCAACGAGGTTTTTTCTGATACCGAAAATGTTGAAAGCTCCATTGCTGGCTCAAAAACACTCGTTGGTGCTCTACCTTATATGGCCCCAGAAATTGTTTTAAACAAAACAAAAGCTGGAAAACATATTGATATTTGGTCAATAGGTGCAATTATGTATTTCCTACTAACAGGAAAAAACACCTTCACATCGCAATTT GCCCAAATCGTCATTAACTATCATACTCAGAAATCTATCGATCCAATCCTTCATATGGATACATCTCGCCATTTAAACCCGTTAGGCAATCAATTACTAGGCATTATCAAAAGTTGCTTGGATTACGACTATTCAAATCGTCCAAAATTCTGAGCAATTAGTCCCTTTGTTACCCCTATTCAAGAAAGAAAATATGGACATATCAAGTATCGACGAGGC ACACATGGTTGGGGCTTCATTAAAAACATAGGCCCTAACGATACATTTTATCATACTGAAGAAGTTTTTGGACTTCAGGCTTCCCATAGTGAGCGAGTGTGCTTCTCTGAGTACCCTGGATTACCTCAAAGCAAGGGCATTTCCAATTATTCGCTGTAAATAA.

Claims (9)

1. A novel anti-phage element, characterized in that the nucleotide sequence is such as SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 or SEQ ID NO.6 shown. 2. A chassis cell containing the novel anti-phage element of claim 1. 3. The chassis cell according to claim 2, characterized in that, the chassis cell is Escherichia coli, pathogenic bacteria Pseudomonas aeruginosa, Salmonella or Vibrio. 4. The application of the novel anti-phage element according to claim 1 in improving the anti-phage infection of chassis cells. 5. The application according to claim 4, characterized in that the chassis cells are Escherichia coli, pathogenic bacteria Pseudomonas aeruginosa, Salmonella or Vibrio. 6. The application according to claim 4, wherein the phage is phage PAP8, PAO-L5, QDWS, PAP-L5, coliphage T1, T4, T5, T7, EEP, λ or M13. 7. A method for improving the anti-phage infection of chassis cells, characterized in that the novel anti-phage elements according to claim 1 are transferred into chassis cells to enhance the host's resistance to phages. 8. The method according to claim 7, characterized in that, the chassis cells are Escherichia coli, pathogenic bacteria Pseudomonas aeruginosa, Salmonella or Vibrio. 9. The method according to claim 7, wherein the phage is phage PAP8, PAO-L5, QDWS, PAP-L5, coliphage T1, T4, T5, T7, EEP, λ or M13.

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