KR101023995B1 - Escherichia coil specific bactriophage and method of preventing or controlling E. coli contamination using the same - Google Patents

Abstract

The present invention Escherichia coli ) relates to the use of said bacteriophage to effectively prevent or control contamination of E. coli in fermentation by cultivation of novel bacteriophages and microorganisms having specific infection and killing ability.

Bacteriophage, Escherichia Coli Contamination, Amino Acids

Description

Escherichia coil specific bactriophage and method of preventing or controlling E. coli contamination using the same}

The present invention Escherichia coli ) relates to the use of said bacteriophage to effectively prevent or control contamination of E. coli in fermentation by cultivation of novel bacteriophages and microorganisms having specific infection and killing ability.

Bacteriophage is a type of bacterial virus discovered by French bacteriologist d'Herelle in 1917, also called phage. Bacteriophage are infected with bacteria to survive and proliferate only within their cells, and the relationship between the bacteriophage and its host is known to have high specificity. Like other viruses, bacteriophages are simple organisms with a protein envelope wrapped around the core of a genetic material. Nucleic acids consist of DNA or RNA, either single- or double-stranded. When infecting a bacterium, the phage cling to the surface of the bacterium and inject its genetic material into the cell, after which the phage follows a lytic or lysogenic life cycle. Soluble phages allow the host cell mechanisms to build their own structures and then destroy or lyse the cells to release new phage particles. Lysogenic phage are incorporated into the host cell's chromosome and replicate with the cell without destroying the cell. Under certain conditions, lysogenic phage can be converted to lytic.

As an antimicrobial drug, lytic bacteriophages can effectively treat infections caused by bacteria because of their ability to replicate internally after infection with sensitive bacteria, creating new bacteriophages that destroy them completely and continue to attack and destroy bacterial cells. Expected. This process can occur both in vitro and in vivo. Bacteriophage, however, was not as effective in killing bacteria in vitro as the effects of killing bacteria in vitro, and the development of antibiotics in the 1940s led to a downhill study of bacteriophages in the West. In some countries, such as Eastern Europe and India, research has been conducted steadily, and recently, due to the misuse of antibiotics, antibiotics resistant to the emergence of antibiotics have attracted much attention.

Existing drugs for the prevention and treatment of diseases using bacteriophage (Atterbury et al., Appl. Envion.Microbiol. 73 (14): 4543-4549 (2007)), disinfectants or cleaning agents to prevent abnormal rot of food (PCT 1998-08944 , PCT 1995-31562, EP 1990-202169, PCT 1990-03122), or the development of a Fiji display technology for diagnosis (Ripp et al., J. Appl. Microbiol. 100 (3): 488-499 (2006) There was no case where the bacteriophage was applied to mass fermentation to prevent bacterial contamination.

In the cultivation of microorganisms to produce useful substances such as amino acids, nucleic acids and the like, contamination by unwanted bacteria causes a decrease in the production efficiency by culturing and, in serious cases, interruption of the culture. Therefore, prevention and control of bacterial contamination in fermentation by microbial culture is very important.

Corynebacterium strains, particularly Corynebacterium glutamicum, are Gram-positive bacteria that are widely used for L-amino acid production, and are used as strains for producing most L-amino acids produced by fermentation. . L-amino acids, in particular L-lysine, is an essential amino acid and is used in fields such as feed, medicine and food. Since L-lysine is mainly produced by a direct fermentation method using Corynebacterium glutamicum, the productivity improvement of L-lysine by the improvement of fermentation process has a great economic effect. In order to improve the fermentation process, mainly attempted to improve the strain or the development of the medium, but the improvement of the L-amino acid production method through microbial culture is still required.

Accordingly, the present inventors have tried to minimize the loss in fermentation by culturing microorganisms other than Escherichia coli, such as Corynebacterium, to maximize the fermentation efficiency. The present invention has been completed by developing a method for effectively preventing or controlling contamination by E. coli, which may occur during commercial mass fermentation.

It is an object of the present invention to provide novel bacteriophage with E. coli specific infection and killing ability.

Still another object of the present invention is to provide a method for effectively preventing or controlling E. coli contamination in large-scale fermentation by culturing microorganisms using a novel bacteriophage having E. coli specific infection and killing ability.

In order to achieve the above object, the present invention has E. coli-specific infection and killing ability, the tail protein having the amino acid sequence of SEQ ID NO: 1 and the base sequence represented by SEQ ID NO: 2 to SEQ ID NO: 37 It provides a bacteriophage belonging to the Siphoviridae family, T1-like phage genus, characterized by having a genome comprising a constructed nucleic acid molecule.

In one embodiment of the invention, the bacteriophage having E. coli specific infection and killing ability may be bacteriophage YS01 KCCM10947.

In one embodiment of the present invention, the bacteriophage may be isolated from the E. coli fermentation broth infected with the bacteriophage.

Figure 1 shows an electron transmission micrograph of the bacteriophage YS01 according to an embodiment of the present invention. According to this, the bacteriophage isolated in the present invention belongs to the siphoviridae family of morphotype B1 in morphology, and the tail having the amino acid sequence of SEQ ID NO: 1 was analyzed through partial sequencing. T1-like phage showing high similarity to bacteriophage RTP, characterized by having a tail protein.

In one embodiment of the present invention, the bacteriophage is specifically infected only with E. coli, and its growth was found to be temperature dependent. When one bacteriophage was infected with one bacterium and self-replicated in it, the burst size was more than 1,000. It showed an infection. Thus, the bacteriophage may be suitable for use in the fermentation industry to effectively prevent or control E. coli contamination.

The present invention also comprises the steps of i) seed culture of microorganisms that are not Escherichia coli, and

ii) a method of preventing or controlling contamination of E. coli in fermentation by culturing the microorganism, comprising inoculating the cultured microorganism with the bacteriophage having E. coli specific infection and killing ability. to provide.

Method for preventing or controlling the contamination of E. coli in fermentation by culturing the microorganism according to the present invention includes the step of culturing the microorganisms that are not E. coli.

In one embodiment of the present invention, the microorganism that is not E. coli may be a microorganism of the genus Corynebacterium (Corynebacterium) having the ability to produce L-amino acids.

 A method for preventing or controlling contamination of E. coli in fermentation by culturing microorganisms according to the present invention includes inoculating the culture medium with the bacteriophage having E. coli specific infection and killing ability. do.

Most of the pollution in most fermentation industries occurs at the time of transition from seed culture to main culture. At this point, a bacteriophage with E. coli-specific infection and killing ability can be added as a means to control E. coli, which has a greater chance of opportunistic infection than other bacteria.

In one embodiment, the bacteriophage has a genome comprising a tail protein having an amino acid sequence of SEQ ID NO: 1 and a nucleic acid molecule represented by a nucleotide sequence consisting of SEQ ID NO: 2 to SEQ ID NO: 37. It may be a bacteriophage belonging to the genus Siphoviridae family, T1-like phage genus.

In one embodiment of the present invention, the bacteriophage may be bacteriophage YSO1 having accession number KCCM10947.

In one embodiment of the present invention, the bacteriophage may be inoculated at a concentration of 10 ^ 4 pfu to 10 ^ 6 pfu per 1L of the culture medium.

In general, considering that the concentration of bacteriophage obtained during the culture is 10 ^ 9 pfu / ml, the amount of 10 ^ 4 pfu to 10 ^ 6 pfu added per 1L of the culture medium is very small. E. coli contamination can be prevented and controlled in L-amino acid fermentation using bacteriophage, which can be inexpensively cultured and isolated.

 In one embodiment of the present invention, the temperature of the culture medium for inoculating the bacteriophage may be 30 ℃ to 33 ℃, preferably 30 ℃.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

In the present invention, it is possible to effectively control the contamination by Escherichia coli without the use of antibiotics in the L-amino acid fermentation process using microorganisms other than Escherichia coli, especially Corynebacterium genus, through the discovery of a novel bacteriophage. In addition, while using a very small amount of bacteriophage, the effect of preventing and controlling E. coli contamination is economical.

Example 1  : Infected with E. coli Bacteriophage  Isolation and Cultivation

(1) Isolation of bacteriophage infected with E. coli

1% chloroform was added to the bacteriophage-infected E. coli fermentation broth, followed by well mixing for 2 minutes, and the supernatant obtained by centrifugation at 12,000 rpm for 10 minutes was filtered using a 0.45 μm filter. The filtrate obtained above was spread over a LB (Luira Bertani) plate (10 g of tryptone; 5 g of yeast extract; 10 g of NaCl; 20 g of agar; H ₂ O in a total volume of 1 L), followed by the culture of E. coli W3110 (OD ₆₀₀ = 0.8) was poured into 0.7% agar medium (agar 7g; H ₂ O in a total volume of 1 L), poured and solidified, and incubated at 30 ° C. for 18 hours. Take one lysate with a wide-pore pippet and fill 500 L SM buffer (NaCl 5.8 g; MgSO ₄ 7H ₂ O 2 g; 1 M Tris-Cl (pH 7.5) 50 ml; fill total volume to 1 L) H ₂ O) and left at room temperature for 6 hours. This was centrifuged to take a supernatant containing bacteriophage infected with E. coli.

(2) Cultivation of bacteriophage infected with E. coli

10 ul of the supernatant prepared in (1) was mixed with 14 ml of 0.7% agar medium and ₅₀₀ ul of E. coli W3100 culture medium (OD ₆₀₀ = 0.8), poured into a 50 mm diameter LB plate, hardened, and incubated at 30 ° C. for 18 hours. 15 ml of SM buffer was dispensed onto the plate with complete lysis and shaken for 5 hours at room temperature. Then, the SM buffer solution was recovered from the plate, and 1% chloroform was added and mixed well for 2 minutes, followed by centrifugation at 12,000 rpm for 10 minutes. The supernatant obtained here was filtered through a 0.45 μm filter and refrigerated.

All experiments were used by repeated culture propagation by the method described in (2) above. The concentration of bacteriophage obtained by the culture was 1.0 * 10 ^ 9 pfu / ml.

Example  2: Bacteriophage  Pure separation and shape observation

Mass culture was performed to purely isolate the bacteriophage. E. coli W3110 was inoculated in LB medium (tryptone 10g; yeast extract 5g; NaCl 10g; H ₂ O in a total volume of 1L), and then incubated at 30 ° C. for 16 hours to obtain 1.0 x 10 ^ 10 cfu per tube. Dispensed as much as possible. After centrifugation at 4000g for 10 minutes it was resuspended in 3ml SM buffer. The bacteriophage obtained in Example 1 was inoculated with 5.0 * 10 ^ 8 pfu and incubated at 30 ° C. for 20 minutes. This was inoculated with two test tubes in 200 ml fresh LB and shaken at 30 ° C. for 8 hours. Then, 4 ml of chloroform was added and shaken for 10 minutes. DNase I and RNase A were inoculated to a final concentration of 1ug / ml and allowed to stand at room temperature for 30 minutes. After addition of 1M NaCl and 10% PEG, the mixture was allowed to stand at 4 ° C for 3 hours. The supernatant was removed after centrifugation for 10 minutes at 4 ℃, 11,000g. The precipitate obtained in the centrifugation was resuspended in 4 ml SM buffer with a wide pore pipette and allowed to stand at room temperature for 20 minutes. After adding 4ml chloroform, the mixture was mixed well, centrifuged at 4 ° C. and 3,000 g for 15 minutes, and the supernatant was separated. The bacteriophage was purified by ultracentrifugation (35,000 rpm, 1 hour, 4 ° C.) using a glycerol density gradient method (density: 40%, 5% glycerol).

The purified bacteriophage was diluted in 0.01% gelatin solution and then fixed with 2.5% glutaraldehyde solution. It was dropped on a carbon-coated mica plate (about 2.5 * 2.5 mm) and acclimated for 10 minutes, followed by washing with sterile distilled water. The carbon film was placed on a copper grid and dyed for 30-60 seconds with 4% uranyl acetate, dried, and examined at 120,000 to 200,000 times magnification at 80 kV with a JEM-1011 transmission electron microscope. 1 shows electron transmission micrographs of bacteriophage isolated and purified through Examples 1 and 2. FIG. As a result, it was confirmed that the isolated bacteriophage belongs to the Siphoviridae family in morphological classification, and named it YS01. Bacteriophage YS01 was deposited on May 15, 2008 with the accession number KCCM10947 to the Korea Microbiological Conservation Center, which is affiliated with the Korea Spawn Association.

Example  3: Bacteriophage  Genetic Characterization

(1) gDNA Isolation of Bacteriophage

In Example 2 gDNA of the purified bacteriophage was extracted by ultracentrifugation. Specifically, the purified bacteriophage culture medium was added with a final concentration of 20 mM EDTA (pH 8.0), 50 ug / ml Protease K, and 0.5% SDS. After mixing well, the mixture was allowed to stand at 56 ° C. for 1 hour. The same amount of phenol (pH 8.0) was added and mixed well, and then centrifuged at 3,000 g for 5 minutes at room temperature. The supernatant was taken and the same amount of PC (phenol: chloroform = 1: 1) was added and mixed well, followed by centrifugation at 3,000 g for 5 minutes at room temperature. After the centrifugation, the supernatant was taken, the same amount of chloroform was added and mixed well, followed by centrifugation at 3,000 g for 5 minutes at room temperature. After taking the supernatant, 3M sodium acetate was added at a ratio of 1/10, mixed, and then 2 times the volume of cold EtOH was added, and the mixture was allowed to stand at -20 ° C for 1 hour. After centrifugation at 12,000 rpm for 10 minutes at 0 ° C, the supernatant was completely removed, and the gDNA precipitated at the bottom was dissolved in 100ul of water and then stored at -20 ° C.

(2) sequencing

To investigate the genetic characteristics of the isolated bacteriophage 5 g of the bacteriophage gDNA obtained in the above (1) was treated with StuI and SmaI restriction enzymes simultaneously. As a vector, pBluescript SK + (Stratagene) was used and cut with EcoR V restriction enzyme and then treated with Cal Intestinal Phosphatase (CIP). The ligation was carried out at 16 ° C. for 5 hours to adjust the reaction conditions such that the amount of gDNA and vector treated as described above was 3: 1. The recombinant vector thus obtained was introduced into E. coli DH5α. Transformants by vectors containing gDNA fragments of bacteriophage were screened only by white colonies on LB plates containing ampicillin and X-gal. Colonies selected by this were shaken at 200 rpm for 16 hours at 30 ° C. in LB medium containing ampicillin. The plasmid was extracted from the E. coli cultured above using a plasmid purification kit (Promega). The size was confirmed by treatment with EcoR V restriction enzyme, and the size was 1 kb or more, and the base sequence was determined using M13 forward primer and M13 reverse primer (Invitrogen) of SEQ ID NOs: 38 and 39, respectively. Analyzed. The gene sequence thus obtained is represented by SEQ ID NOs: 2 to 37 as a partial sequence constituting the entire genome of the isolated bacteriophage YS01.

The gene sequences obtained above were analyzed using the NCBI blast program, and the results are shown in Tables 1 and 2 below. Sequence analysis revealed that bacteriophage YS01 was a T1-like phage showing high similarity with bacteriophage RTP.

Table 1. Analysis results using blastn

snippet
(SEQ ID NO) organism gene identities Gaps One
(2/3) Enterobacteria phage RTP complete genome 389/558 (69%) 48/558 (8%) 2F
(4) Enterobacteria phage RTP complete genome 93/108 (86%) 0/108 (0%) 2R
(5) Bacteriophage RTP complete genome 582/722 (80%) 0/722 (0%) Bacteriophage JK06 virion complete genome 569/725 (78%) 2/725 (0%) Phage TLS complete genome 462/667 (69%) 9/667 (1%) 3
(6/7) Enterobacteria phage RTP complete genome 88/104 (84%) 0/104 (0%) 4
(8/9) Enterobacteria phage RTP complete genome 149/182 (81%) 0/182 (0%) 6
(12/13) Bacteriophage JK06 virion complete genome 245/331 (74%) 2/331 (0%) Enterobacteria phage RTP complete genome 291/430 (67%) 10/430 (2%) 7
(14/15) Enterobacteria phage RTP complete genome 89/104 (85%) 0/104 (0%) 8
(16/17) Enterobacteria phage RTP complete genome 579/720 (80%) 0/720 (0%) Bacteriophage JK06 virion complete genome 565/722 (78%) 2/722 (0%) Bacteriophage TLS complete genome 457/662 (69%) 9/662 (1%) Enterobacteria phage T1 complete genome 467/709 (65%) 22/709 (3%) 10
(20/21) Enterobacteria phage RTP complete genome 422/606 (69%) 48/606 (7%) Bacteriophage JK06 virion complete genome 98/131 (74%) 8/131 (6%) 11
(22/23) Bacteriophage RTP complete genome 297/346 (85%) 0/346 (0%) Bacteriophage jk06 complete genome 272/347 (78%) 2/347 (0%) Phage TLS complete genome 264/376 (70%) 11/376 (2%) 12
(24/25) Bacteriophage RTP complete genome 149/182 (81%) 0/182 (0%) 13
(26/27)
Bacteriophage RTP complete genome 422/606 (69%) 48/606 (7%) Bacteriophage jk06 complete genome 95/124 (76%) 3/124 (2%) 14
(28/29) Bacteriophage RTP complete genome 89/104 (85%) 0/104 (0%)

Note: F and R written in the number of fragments mean Forward and Reverse, respectively. That is, SEQ ID 2F shows the result read using M13 forward primer, and SEQ ID 2R shows the result read using M13 reverse primer. In addition, when two sequence numbers are described for a fragment, the result of reading the forward / reverse primer corresponds to one fragment.

Table 2. Analysis results using blastx

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(SEQ ID NO) organism protein identities Positive One
(2/3) Bacteriophage RTP hypothetical protein rtp26 90/171 (52%) 101/171 (59%) Enterobacteria phage T1 hypothetical protein T1p49 46/93 (49%) 62/93 (66%) 2F
(4) Bacteriophage TLS TtpM 116/236 (49%) 156/236 (66%) Enterobacteria phage T1 putative tail tape protein 101/236 (42%) 155/236 (65%) 2R
(5) Bacteriophage TLS YfiJ 176/241 (73%) 208/241 (86%) 4
(8/9) Enterobacteria phage RTP hypothetical protein rtp15 59/61 (96%) 60/61 (98%) 6F
(12) Bacteriophage jk06 putative tail tape protein 139/249 (55%) 178/249 (71%) Enterobacteria phage RTP putative tail tape protein 96/161 (59%) 123/161 (76%) 6R
(13) Bacteriophage jk06 putative minor tail protein 78/111 (70%) 90/111 (81%) Enterobacteria phage RTP putative minor tail protein 73/113 (64%) 88/113 (77%) 8
(16/17) Enterobacteria phage RTP hypothetical protein rtp67 215/240 (89%) 228/240 (95%) Bacteriophage TLS YfiJ 175/240 (72%) 207/240 (86%) 11F
(22) Enterobacteria phage RTP hypothetical protein rtp69 80/93 (86%) 90/93 (96%) Bacteriophage jk06 hypothetical protein JK_50 66/93 (70%) 78/93 (83%) 11R
(23) Enterobacteria phage RTP hypothetical protein rtp67 111/116 (95%) 113/116 (97%) Bacteriophage jk06 hypothetical protein JK_52 102/117 (87%) 110/117 (94%)

Note: F and R written in the number of fragments mean Forward and Reverse, respectively. That is, SEQ ID NO: 2F shows the result read using M13 forward primer, and SEQ ID 2R shows the result read using M13 reverse primer. In addition, when two sequence numbers are described for a fragment, the result of reading the forward / reverse primer corresponds to one fragment.

Example  4: Bacteriophage YS01 Range of infection of E. coli Corynebacterium  Glutamicum ( Corynebacterium glutamicum ) And Salmonella spp. ( Salmonella spp .) Degree of infection compare)

The lytic activity of bacteriophage YS01 against bacteria other than Escherichia coli, namely Salmonella and Corynebacterium bacteria, was investigated. Each bacterium was incubated to OD ₆₀₀ = 0.8 and then mixed in 0.7% agar medium and poured into LB plates to harden. Bacteriophage YS01 was diluted 10-fold from 10 ^ 8 pfu to 10 ^ 3 pfu using SM buffer, and each diluted solution was dropped by 10ul on the LB plate, and then cultured at 30 ° C. for 16 hours to observe whether plaque was formed. It was. The bacteria used in this experiment and their infection are listed in Table 3 below.

 Table 3. Infection Range Survey of Bacteriophage YS01

Strain name Whether plaque is produced E. coli (W3110) O E. coli (MG1655) O E. coli (BL21) O E. coli (DH5α) O Salmonella Galina Room (Salmonella gallinarum ) X Salmonella pool room (Salmonella pullorum ) X Salmonella typhimurium (Salmonella typhimurium ) X Salmonella Entebbe utility disk (Salmonella enteritidis ) X Derby Salmonella (Salmonella derby ) X Corynebacterium glutamicum (Corynebacterium glutamicum ) X

Example  5: Bacteriophage YS01 One-step growth curve of One - step growth  curve) (Infection Test)

The infectivity of bacteriophage YS01 according to the culture temperature was investigated. E. coli W3110 was incubated in LB medium at OD = 0.5 at 30 ° C. and 37 ° C., respectively, and then concentrated twice. The bacteriophage YS01 isolated in the concentrated E. coli culture was inoculated with multiplicity of infection (MOI) = 0.0005 and allowed to stand for 5 minutes. The reaction solution was centrifuged to separate the cells and then suspended again in fresh LB medium. Thereafter, two samples were taken from the culture solution at 30 ° C. and 37 ° C., 5 min intervals up to 30 minutes, and 10 min intervals up to 80 minutes, respectively, one treated with chloroform and the other not treated. 2 shows the one-step growth curve of bacteriophage YS01 incubated at 30 ° C. Diluting the culture solution obtained by the above culture step by step 10 ml of E. coli (W3110) culture solution (OD ₆₀₀ = 0.8) in 0.7% agar medium, poured into a plate and dropped on the hardened LB plate, respectively at 30 ℃ and 37 ℃ It was incubated for 18 hours and observed for lysis. As a result, bacteriophage YS01 was found to cause a smooth infection at 30 ℃ than 37 ℃, it was found that the discharge water (burst size) at 30 ℃ or more than 1,000. Figure 3 shows the infectivity according to the culture temperature of bacteriophage YS01.

Example  6: Escherichia coli W3110 For Bacteriophage YS01 Bacteriostatic ability of clearance ) analysis

The bacteriostatic experiment was carried out under various conditions to determine bacteriostatic capacity against E. coli on the liquid medium of bacteriophage YS01. After putting 35 ml of LB liquid medium in a 250 ml flask, W3110: YS01 was inoculated under the conditions of 1: 1, 500: 1, 1,000: 1, 10,000: 1, 100,000: 1, 500,000: 1, and 10,000,000: 1, respectively. After inoculation, shaking culture was carried out at 30 ° C. at 200 rpm, and the change in OD over time was measured. Compared to the negative control group not inoculated with bacteriophage YS01, the experimental group showed excellent bacteriostatic ability after 2 hours even under the conditions of W3110: YS01 = 10,000: 1, and it was confirmed that the bacteriostatic ability was also achieved under the condition of 10,000,000: 1. FIG. 4 is a clearance curve showing bacteriostatic capacity against various proportions of bacteriophage YSO1 against host cells. FIG.

Example  7: Prevention of E. coli infection during lysine fermentation Application example

L-lysine was produced by culturing Corynebacterium glutamicum KFCC10881 to evaluate the effect of bacteriophage YSO1 on lysine fermentation using Corynebacterium.

First, 25 ml of seed medium (20 g of glucose, 10 g of peptone, 5 g of yeast extract, 1.5 g of urea, KH ₂ PO _{4 4} g, K ₂ HPO ₄ 8 g, MgSO ₄ 7H ₂ O 0.5 g, biotin 100 µg, thiamine HCL 1000 µg, calcium Inoculate Corynebacterium glutamicum strain KFCC10881 into a 250 ml corner-baffle flask containing 2000 μg pantothenic acid, 2000 μg nicotinamide: pH 7.0, per liter of process water) and at 200 rpm for 72 hours at 30 ° C. The culture medium was prepared by incubating with stirring. Then, the culture fermentation medium (glucose 50g, (NH ₄ ) ₂ SO ₄ 40g, soy protein 2.5g, Corn Steep Solids 5g, urea 3g, KH ₂ PO ₄ 1g, MgSO ₄ 7H ₂ O 0.5 g, biotin 100 µg, thiamine HCL 1000 µg, calcium-pantothenic acid 2000 µg, nicotinamide 3000 µg, CaCO ₃ 30 g: pH 7.0, based on 1 liter of the process water) and inoculated the species culture solution with bacteriophage YS01 at a constant concentration. After addition, the effects of bacteriophage YS01 on E. coli infection and fermentation were examined. At the time of inoculation, E. coli W3110 and bacteriophage YS01 were inoculated in the same ratio as described in Table 4 below. For comparison, the effect of E. coli W3110 or bacteriophage YS01 was inoculated alone. Two conditions of high and low titers were tested for E. coli W3110 and bacteriophage YS01, respectively. As shown in Table 4, the group infected with E. coli W3110 alone showed about 54% L-lysine production at the end of fermentation compared to the normal control group that had not been inoculated with E. coli or bacteriophage at all, resulting in a negative effect on L-lysine fermentation of E. coli W3110. In the group inoculated with bacteriophage YS01 alone, it was confirmed that the bacteriophage YS01 alone did not affect lysine fermentation because there was no change in L-lysine production compared to the normal control group. In the co-administered group inoculated with E. coli W3110 and bacteriophage YS01, the experimental group with E. coli W3110: bacteriophage YS01 ratio of 10,000: 1 showed almost the same amount of L-lysine production as the normal control group. 5 shows the effect on L-lysine fermentation by E. coli W3110, bacteriophage YS01 and combinations thereof.

Table 4. Assessment of the impact of YS01 on L-lysine fermentation

Lysine
Production strain Condition W3110
( cfu ) YS01
( cfu ) L-lysine (g) Corynebacterium glutamicum KFCC10881 Negative Control (N.C.) x x 18.78 YS01 (high potency) x 1.0 * 10 ^ 8 18.86 YS01 (low titer) x 6.7 * 10 ^ 5 18.67 W3110 (high titer): YS01 = 150: 1 1.0 * 10 ^ 8 6.7 * 10 ^ 5 18.34 W3110 (high titer): YS01 = 10,000: 1 1.0 * 10 ^ 8 1.0 * 10 ^ 4 18.82 W3110 (low titer): YS01 = 150: 1 1.0 * 10 ^ 7 6.7 * 10 ^ 4 18.36 W3110 (low titer): YS01 = 10,000: 1 1.0 * 10 ^ 7 1.0 * 10 ^ 3 18.35 Positive control (high titer) 1.0 * 10 ^ 8 x 10.22 Positive control (low titer) 1.0 * 10 ^ 7 x 12.30

1 is an electron transmission micrograph of bacteriophage YS01 having E. coli specific infection and killing ability isolated from the present invention.

2 shows the one-step growth curve of bacteriophage YS01 incubated at 30 ° C.

3 shows infectivity according to temperature of bacteriophage YS01.

4 shows the clearance curve of bacteriophage YSO1. N.C. represents a negative control.

5 shows the effect on L-lysine fermentation by E. coli W3110, bacteriophage YS01 and combinations thereof in a method according to one embodiment of the invention.

<110> CJ Corporation <120> Escherichia coil specific bactriophage and method of preventing          or controlling E. coli contamination using the same <160> 39 <170> KopatentIn 1.71 <210> 1 <211> 236 <212> PRT <213> Bacteriophage YS01 <220> <221> PEPTIDE (222) (1) .. (236) <220> <221> PEPTIDE (222) (1) .. (236) <223> tail protein <400> 1 Lys Val Asn Asp Ile Gly Thr Ser Leu Ala Gly Gly Ile Pro Ile Trp   1 5 10 15 Leu Ile Ala Ile Gln Gln Gly Gly Gln Ile Lys Asp Ser Phe Gly Gly              20 25 30 Ile Ala Asn Thr Ala Arg Phe Ala Leu Ala Ala Ile Asn Pro Leu Thr          35 40 45 Ala Val Val Gly Thr Leu Gly Leu Ala Leu Gly Ala Leu Gly Val Ala      50 55 60 Ala Tyr Gln Ala Asp Lys Asn Thr Arg Gly Leu Ala Ser Thr Ile Thr  65 70 75 80 Leu Val Gly Asn Ser Thr Ile Thr Ser Thr Ala Gln Ile Asn Gln Val                  85 90 95 Val Asp Gly Ile Glu Arg Ser Thr Leu Ala Thr Arg Gly Leu Ile Gln             100 105 110 Glu Ile Ala Asn Ser Leu Val Ala Asn Gly Asn Leu Thr Leu Thr Gln         115 120 125 Ile Asp Lys Ile Thr Lys Ala Thr Ala Gln Trp Ala Thr Val Thr Ser     130 135 140 Thr Asp Ser Lys Thr Ile Leu Gly Tyr Phe Asp Gln Ile Thr Lys Asp 145 150 155 160 Pro Ile Gln Gly Leu Ile Ala Leu Asp Lys Gln Phe Asn Phe Leu Gln                 165 170 175 Ser Gly Gln Leu Lys Tyr Ile Glu Ser Ile Arg Lys Thr Ala Gly Glu             180 185 190 Thr Glu Ala Val Thr Ala Ala Thr Glu Leu Phe Ala Asn Val Met Glu         195 200 205 Asn Arg Leu Ala Asp Val Ala Leu Ser Leu Asn Pro Leu Glu Ser Ala     210 215 220 Trp Asn Ser Phe Arg Lys Phe Val Ser Glu Ile Trp 225 230 235 <210> 2 <211> 766 <212> DNA <213> Bacteriophage YS01 <400> 2 atcagtgata cttaggtctg tgccggatac cattaaacac ctgacaggta aatcacctac 60 cagaataata ctgtcgtaag gctggaatgc tggaatcaag gcctttgctt tttactcgag 120 gccttcgcaa ctatttgaag cgatttcaac taactccatt tgaatctcct gattatttat 180 ggtgcctacc attctatcag ataaaatgca ataaaaaacc tcccgaagga ggcttattta 240 attaacgcca aacttatttc tgtttaacct gaacctctac cagaatgcca tcaacactat 300 cgtcttgagt tggcattgcg ccggctgctg ttgctgctgc gatatagcct ccagcaaaag 360 tccaaccagt agtgaaagct gccgcaccag cgactgctgc atcaacaacc aaacctgcgg 420 cattgataaa tactggttta ccaacggctc ggtttgcggc ggtaattgct gctgctgccc 480 ttacccagat acgaccagca gtcattacgt taacagcctc ctgagctttg gctgtgccat 540 ctggagtctc ataatgagaa cgaacaacaa caccgtaagc gctaccttct tcatttactg 600 cggtaacaac cttgtgacca tcaatctcag gagcggtagt tacggaaacc gccacaccaa 660 cagggattgc atcagagcca ccaactacac aagcgccatc gatattatac agggaagtat 720 cagctaccat acctgccagt gctacagaac gaattgccat ttatct 766 <210> 3 <211> 747 <212> DNA <213> Bacteriophage YS01 <400> 3 atcattgacc ttgacccgcg cgcttgcctg aagaaaatta aaaagtaagg agatagataa 60 atggcaattc gttctgtagc actggcaggt atggtagctg atacttccct gtataatatc 120 gatggcgctt gtgtagttgg tggctctgat gcaatccctg ttggtgtggc ggtttccgta 180 actaccgctc ctgagattga tggtcacaag gttgttaccg cagtaaatga agaaggtagc 240 gcttacggtg ttgttgttcg ttctcattat gagactccag atggcacagg caaagctcag 300 gaggctgtta acgtaatgac tgctggtcgt atctgggtaa gggcagcagc agcaattacc 360 gccgcaaacc gagccgttgg taaaccagta tttatcaatg ccgcaggttt ggttgttgat 420 gcagcagtcg ctggtgcggc agctttcact actggttgga cttttgctgg aggctatatc 480 gcagcagcaa cagcagccgg cgcaatgcca actcaagacg atagtgttga tggcattctg 540 gtagaggttc aggttaaaca gaaataagtt tggcgttaat taaataagcc tccttcggga 600 ggttttttat tgcattttat ctgatagaat ggtangcacc ataaataatc angagattca 660 natggagtta gttgaaatcg cttcaaatag ttgcgaaagc ctcgagtaan aaagcaaggc 720 cttgattcca gcattccagc cttacga 747 <210> 4 <211> 772 <212> DNA <213> Bacteriophage YS01 <400> 4 atcatcccag atctcactca caaacttcct gaagctattc catgcacttt caagaggatt 60 gaggcttagc gcaacgtccg caagcctgtt ttccataacg ttagcgaata actcagtagc 120 tgcggttact gcctctgttt ctccagccgt cttcctgatg ctctcaatgt atttaagctg 180 cccggactga aggaagttaa attgcttatc aagcgcaatc aagccttgta tcgggtcttt 240 tgtgatttga tcaaagtacc caagtattgt ctttgaatcc gttgatgtta cagttgccca 300 ctgagcagtt gctttagtta tcttatctat ctgcgtaagc gttaggttgc cgtttgctac 360 aaggctgttc gctatctcct gaataagacc gcgagtagct agtgttgatc tctcaattcc 420 atcaacaacc tgatttatct gcgctgtaga cgtgatagtt gagttgccaa ccaacgttat 480 cgtagatgct agtcctcgag tattcttgtc tgcctggtaa gctgcaacac caagagcgcc 540 aagagctaaa cccaaagtcc caacaactgc tgttaatggg ttaattgctg ctaatgcaaa 600 ccttgcggta ttagcaatgc caccaaaact atccttaatc tgaccgccct gctgaattgc 660 aatgagccag attgggatgc caccagcaag ggaagttccg atatcattga ccttgacccg 720 cgcgcttgcc tgaagaaatt aaaaagtaag gagatagata aatggcaatt cc 772 <210> 5 <211> 733 <212> DNA <213> Bacteriophage YS01 <400> 5 atctgacgaa gcaacaagcg taaactcact gcttggcatg acgtatggcg atggctctaa 60 gaaaacgaac agcgaactcg tccttaaagc ctgggatgct ggtaacgtgt ccattgcgcg 120 tgcgaacgca gaaaacaata tgagcttcgt agctatggga tgtatctctg taattgccca 180 ggatgaaaca attaacgcca tcatgaatgc cggtgctcgt ggtattggtg tgtcggaacg 240 ttttcttctg gtgcgtgaaa agtcatttct tggcgatcgc gtgtttgttg atgagaatgg 300 taattcaaca tatgaggcta ttgatggtgg gctaaaggct gattacttcc gactcgttca 360 tgagattatg aatgaagtag aagtgaaact aactgtaagt gcttcttcaa tgcgttacct 420 gaaccgtgcg cgccaggaaa tggagccgca tcttggtgac ggaggtaagt attctcacac 480 aatgttgcgc ggtgctctag gtaagtttga taagcaagca atccgtatcg cttctgttct 540 gcacgttatt cgtaactggt tcgcaccagc aggctcaagc cctcataaat ccagagagat 600 tgaactggag acaatgcang aagcggttat gattttcaac gaactaagta aaacttattt 660 gtcatctgca aacgctgctg gtcacgctgg tgacaatgct gagatgaata agctgattga 720 tatcagtgat act 733 <210> 6 <211> 702 <212> DNA <213> Bacteriophage YS01 <400> 6 atcatcccag atctcactca caaacttcct gaagctattc catgcacttt caagaggatt 60 gaggcttagc gcaacgtccg caagcctgtt ttccataacg ttagcgaata actcagtagc 120 tgcggttact gcctctgttt ctccagccgt cttcctgatg ctctcaatgt atttaagctg 180 cccggactga aggaagttaa attgcttatc aagcgcaatc aagccttgta tcgggtcttt 240 tgtgatttga tcaaagtacc caagtattgt ctttgaatcc gttgatgtta cagttgccca 300 ctgagcagtt gctttagtta tcttatctat ctgcgtaagc gttaggttgc cgtttgctac 360 aaggctgttc gctatctcct gaataagacc gcgagtagct agtgttgatc tctcaattcc 420 atcaacaacc tgatttatct gcgctgtaga cgtgatagtt gagttgccaa ccaacgttat 480 cgtagatgct agtcctcgag tattcttgtc tgcctggtaa gctgcaacac caagagcgcc 540 aagagctaaa cccaaagtcc caacaactgc tgttaatggg ttaattgctg ctaatgcaaa 600 ccttgcggta ttagcaatgc caccaaaact atccttaatc tgaccgccct gctgaattgc 660 aatgagccag attgngatgc caccagcaag ggaagttccg at 702 <210> 7 <211> 702 <212> DNA <213> Bacteriophage YS01 <400> 7 atcggaactt cccttgctgg tggcatccca atctggctca ttgcaattca gcagggcggt 60 cagattaagg atagttttgg tggcattgct aataccgcaa ggtttgcatt agcagcaatt 120 aacccattaa cagcagttgt tgggactttg ggtttagctc ttggcgctct tggtgttgca 180 gcttaccagg cagacaagaa tactcgagga ctagcatcta cgataacgtt ggttggcaac 240 tcaactatca cgtctacagc gcagataaat caggttgttg atggaattga gagatcaaca 300 ctagctactc gcggtcttat tcaggagata gcgaacagcc ttgtagcaaa cggcaaccta 360 acgcttacgc agatagataa gataactaaa gcaactgctc agtgggcaac tgtaacatca 420 acggattcaa agacaatact tgggtacttt gatcaaatca caaaagaccc gatacaaggc 480 ttgattgcgc ttgataagca atttaacttc cttcagtccg ggcagcttaa atacattgag 540 agcatcagga agacggctgg agaaacagag gcagtaaccg cagctactga gttattcgct 600 aacgttatgg aaaacaggct tgcggacgtt gcgctaagcc tcaatcctct tgaaagtgca 660 tggaatagct tcaggaagtt tgtgagtgag atctgggatg at 702 <210> 8 <211> 470 <212> DNA <213> Bacteriophage YS01 <400> 8 ccttgattcc agcattccag ccttacgaca gtattattct ggtaggtgat ttacctgtca 60 ggtgtttaat ggtatccggc acagacctaa gtatcactga tatcacccgc atgattgatg 120 cgatcgagac tagcacccaa ggcttgcata ctgccactga tgtagcaatc ccatacgaag 180 atgagatatt ggatgacctt attttcttca atttgctggt tgggcgcgaa ttcacttgtg 240 atggagataa acgcgaagct ggttcaaatt acgaggatgg tgaataatgt ttggattaaa 300 cgacgcatac tttaatgcag ttaagcgcca ggctgataag ctaaacgacg aatacggcaa 360 gctgggtgcg aagaagaaag atgacaaagt tgtggcggca ctgattactc aggtttgggg 420 tccagtttct acgctaatcg cgagggataa gtttgtgtgg atagcgggat 470 <210> 9 <211> 470 <212> DNA <213> Bacteriophage YS01 <400> 9 atcccgctat ccacacaaac ttatccctcg cgattagcgt agaaactgga ccccaaacct 60 gagtaatcag tgccgccaca actttgtcat ctttcttctt cgcacccagc ttgccgtatt 120 cgtcgtttag cttatcagcc tggcgcttaa ctgcattaaa gtatgcgtcg tttaatccaa 180 acattattca ccatcctcgt aatttgaacc agcttcgcgt ttatctccat cacaagtgaa 240 ttcgcgccca accagcaaat tgaagaaaat aaggtcatcc aatatctcat cttcgtatgg 300 gattgctaca tcagtggcag tatgcaagcc ttgggtgcta gtctcgatcg catcaatcat 360 gcgggtgata tcagtgatac ttaggtctgt gccggatacc attaaacacc tgacaggtaa 420 atcacctacc agaataatac tgtcgtaagg ctggaatgct ggaatcaagg 470 <210> 10 <211> 101 <212> DNA <213> Bacteriophage YS01 <400> 10 atcagtgata cttaggtctg tgccggatac cattaaacac ctgacaggta aatcacctac 60 cagaataata ctgtcgtaag gctggaatgc tggaatcaag g 101 <210> 11 <211> 101 <212> DNA <213> Bacteriophage YS01 <400> 11 ccttgattcc agcattccag ccttacgaca gtattattct ggtaggtgat ttacctgtca 60 ggtgtttaat ggtatccggc acagacctaa gtatcactga t 101 <210> 12 <211> 776 <212> DNA <213> Bacteriophage YS01 <400> 12 atcctacttg gtgagtttgc aatagcggca accaagacac ttcagtcaat acctggactt 60 ggcgatttag gtaataaagc aatcgctgag cagcaaaaga ttgtcgatgc tgctaagaag 120 caaaacattg agttagcgaa aagcatagct gagcgagatg ctagaatcag gaagggtgaa 180 cttggttatg ttgagggcaa caataatacc aacctgagcg gcaataacgc atcaagcgat 240 aaggagtttg cagcaaggaa gaaggcgctt gcggatgagt atgatgcgat aaagaaagcg 300 aggagtgaga ggagcaaggc agtaaaagaa gaaagggatt taactttatc ttatgaatct 360 ggtgttcttg ctcttcaggc tcagctgaag gttttacaag agcacaggac gataaacgat 420 gtaataagca acgaacgcaa gcagctattc caggaagaag ctaagtttgc aatacttgag 480 caaaggaaag cggatggcac gcttacaaaa tcacaggcta agctacttgc gcagaagaat 540 atcattcttg aacaagctaa acagaaggct gaacttggtg accagattgt acttcaggag 600 aggtcanaca agttacttga tgacaacctt aagaagactg tgcagattaa gaatgaggca 660 aataatgttt cgcttgctgc taatttatct gacaggnaga agcagagagc aaaagagctt 720 caggcacttc aatcaatcag aataacaaag tggctctgta gatgacactg attttc 776 <210> 13 <211> 715 <212> DNA <213> Bacteriophage YS01 <400> 13 ccttgctcca tactcttcgc cacggaatgt aattgtcttg gctggtagtg actgaccaga 60 tgcctgagct tgcatgattt cctcgggcgt gtatgagatg ttctccccat gaaacctgta 120 aacccttgcg ccgaacttac tgccgtcaac ctcaataagc gtaaccacct cgccagggaa 180 aagtgactga agctggttct caaactttac acttgccata ttctctccaa taaaaaaggc 240 cctcaatggg ccttagttta actcattgac gtgaaaactt cagtaaattg catgctaaca 300 gtctgaactt ctttcgtgct tggcgagacg ctaatgctat cctgctgaat aaaaaacagc 360 tttagttcgc cctgcggtgt agtccatgcg aaaggtttta cgatgtgatc aaagcagaag 420 ttcattaccg cctcccaatc acgaccgacg tagctaacag tatatgtcct cttgttagtc 480 ctgtatccac cagttccgcg ctgctgataa ccattcgcaa atgagacaac cctaacattg 540 ttggcgttag taaatgaaga gcctccattg agaaacattt aacacaaatt taaagttgca 600 atgncattta aatccttagg gcccgaaccc cattaaatag gtgtaccgcg caagaagcgt 660 acgctctgcc gcctgctgga tgatttgctg attcctcagc acgataactt actcc 715 <210> 14 <211> 783 <212> DNA <213> Bacteriophage YS01 <400> 14 atcggaactt cccttgctgg tggcatccca atctggctca ttgcaattca gcagggcggt 60 cagattaagg atagttttgg tggcattgct aataccgcaa ggtttgcatt agcagcaatt 120 aacccattaa cagcagttgt tgggactttg ggtttagctc ttggcgctct tggtgttgca 180 gcttaccagg cagacaagaa tactcgagga ctagcatcta cgataacgtt ggttggcaac 240 tcaactatca cgtctacagc gcagataaat caggttgttg atggaattga gagatcaaca 300 ctagctactc gcggtcttat tcaggagata gcgaacagcc ttgtagcaaa cggcaaccta 360 acgcttacgc agatagataa gataactaaa gcaactgctc agtgggcaac tgtaacatca 420 acggattcaa agacaatact tgggtacttt gatcaaatca caaaagaccc gatacaaggc 480 ttgattgcgc ttgataagca atttaacttc cttcagtccg ggcagcttan atacattgag 540 agcatcagga agacggctgg agaaacagag gcagtaaccg cagctactga gttattcgct 600 aacgttatgg aaaacaggct tgcggacgtt gcgctaagcc tcaatcctct tgaaagtgca 660 tggaatagct tcaggaagtt tgtgagtgag atctgggatg atatcgaaat cctgcagccc 720 gggggatcca ctagntctag agcggccgcc accgcggtga agctccagct tttgttccct 780 tta 783 <210> 15 <211> 701 <212> DNA <213> Bacteriophage YS01 <400> 15 atcatcccag atctcactca caaacttcct gaagctattc catgcacttt caagaggatt 60 gaggcttagc gcaacgtccg caagcctgtt ttccataacg ttagcgaata actcagtagc 120 tgcggttact gcctctgttt ctccagccgt cttcctgatg ctctcaatgt atttaagctg 180 cccggactga aggaagttaa attgcttatc aagcgcaatc aagccttgta tcgggtcttt 240 tgtgatttga tcaaagtacc caagtattgt ctttgaatcc gttgatgtta cagttgccca 300 ctgagcagtt gctttagtta tcttatctat ctgcgtaagc gttaggttgc cgtttgctac 360 aaggctgttc gctatctcct gaataagacc gcgagtagct agtgttgatc tctcaattcc 420 atcaacaacc tgatttatct gcgctgtaga cgtgatagtt gagttgccaa ccaacgttat 480 cgtagatgct agtcctcgag tattcttgtc tgcctggtaa gctgcaacac caagagcgcc 540 aagagctaaa cccaaagtcc caacaactgc tgttaatggg ttaattgctg ctaatgcana 600 ccttgncgta ttagcaatgc caccaaaact atccttaatc tgnaacgcct gctgaattgc 660 atgagccaga ttgggatgcc acccagcagg gaaagtccga t 701 <210> 16 <211> 721 <212> DNA <213> Bacteriophage YS01 <400> 16 atcaatcagc ttattcatct cagcattgtc accagcgtga ccagcagcgt ttgcagatga 60 caaataagtt ttacttagtt cgttgaaaat cataaccgct tcctgcattg tctccagttc 120 aatctctctg gatttatgag ggcttgagcc tgctggtgcg aaccagttac gaataacgtg 180 cagaacagaa gcgatacgga ttgcttgctt atcaaactta cctagagcac cgcgcaacat 240 tgtgtgagaa tacttacctc cgtcaccaag atgcggctcc atttcctggc gcgcacggtt 300 caggtaacgc attgaagaag cacttacagt tagtttcact tctacttcat tcataatctc 360 atgaacgagt cggaagtaat cagcctttag cccaccatca atagcctcat atgttgaatt 420 accattctca tcaacaaaca cgcgatcgcc aagaaatgac ttttcacgca ccagaagaaa 480 acgttccgac acaccaatac cacgagcacc ggcattcatg atggcgttaa ttgtttcatc 540 ctgngcaatt acagagatac atcccatagc tacgaagctc atattgtttt ctgcgttcgc 600 acgcgcaatg gacacgttac cagcatccca ggctttaagg acgagttcgc tgttcgtttt 660 cttagagcca tcgccatacg tcatgccaag cagtgagttt acgcttgntg cttcgtcaga 720 t 721 <210> 17 <211> 721 <212> DNA <213> Bacteriophage YS01 <400> 17 atctgacgaa gcaacaagcg taaactcact gcttggcatg acgtatggcg atggctctaa 60 gaaaacgaac agcgaactcg tccttaaagc ctgggatgct ggtaacgtgt ccattgcgcg 120 tgcgaacgca gaaaacaata tgagcttcgt agctatggga tgtatctctg taattgccca 180 ggatgaaaca attaacgcca tcatgaatgc cggtgctcgt ggtattggtg tgtcggaacg 240 ttttcttctg gtgcgtgaaa agtcatttct tggcgatcgc gtgtttgttg atgagaatgg 300 taattcaaca tatgaggcta ttgatggtgg gctaaaggct gattacttcc gactcgttca 360 tgagattatg aatgaagtag aagtgaaact aactgtaagt gcttcttcaa tgcgttacct 420 gaaccgtgcg cgccaggaaa tggagccgca tcttggtgac ggaggtaagt attctcacac 480 aatgttgcgc ggtgctctag gtaagtttga taagcaagca atccgtatcg cttctgttct 540 gcacgttatt cgtaactggt tcgcaccagc aggctcaagc cctcataaat ccagagagat 600 tgaactggag acaatgcagg aagcggttat gattttcaac gaactaagta aaacttattt 660 gtcatctgca aacgctgctg gtcacgctgg tgacaatgct gagatgaata agctgattga 720 t 721 <210> 18 <211> 342 <212> DNA <213> Bacteriophage YS01 <400> 18 cgagatggtt aagcaagcgc taaagtctga agataagcaa ttcagcgttt cattaactaa 60 gattgaaggg attcttggta atgaagaggg gtcgattaag aagctgaaga gctcgtacat 120 ttataacaca acggctcgta tgcctgagct tagggatgct ggactgaaag ctactcacca 180 ttacgatgaa gatgagtttg aaaatccatt tggtgaaaca attctatctg gagacaagaa 240 gttagttatt aaactggtcc ctggaacggc aaatattggt atcaggaaga agaccaacga 300 cgcaatcaag ttggaggcaa tcactgaatt caaggctaag at 342 <210> 19 <211> 342 <212> DNA <213> Bacteriophage YS01 <400> 19 atcttagcct tgaattcagt gattgcctcc aacttgattg cgtcgttggt cttcttcctg 60 ataccaatat ttgccgttcc agggaccagt ttaataacta acttcttgtc tccagataga 120 attgtttcac caaatggatt ttcaaactca tcttcatcgt aatggtgagt agctttcagt 180 ccagcatccc taagctcagg catacgagcc gttgtgttat aaatgtacga gctcttcagc 240 ttcttaatcg acccctcttc attaccaaga atcccttcaa tcttagttaa tgaaacgctg 300 aattgcttat cttcagactt tagcgcttgc ttaaccatct cg 342 <210> 20 <211> 828 <212> DNA <213> Bacteriophage YS01 <400> 20 actatgggcg attgggtacc gggcccccct cgaggtcatc attgaccttg acccgcgcgc 60 ttgcctgaag aaaattaaaa agtaaggaga tagataaatg gcaattcgtt ctgtagcact 120 ggcaggtatg gtagctgata cttccctgta taatatcgat ggcgcttgtg tagttggtgg 180 ctctgatgca atccctgttg gtgtggcggt ttccgtaact accgctcctg agattgatgg 240 tcacaaggtt gttaccgcag taaatgaaga aggtagcgct tacggtgttg ttgttcgttc 300 tcattatgag actccagatg gcacagccaa agctcaggag gctgttaacg taatgactgc 360 tggtcgtatc tgggtaaggg cagcagcagc aattaccgcc gcaaaccgag ccgttggtaa 420 accagtattt atcaatgccg caggtttggt tgttgatgca gcagtcgctg gtgcggcagc 480 tttcactact ggttggactt ttgctggagg ctatatcgca gcagcaacag cagccggcgc 540 aatgccaact caagacgata gtgttgatgg cattctggta gaggttcagg ttaaacagaa 600 ataagtttgg cgttaattaa ataagcctcc ttcgggaggt tttttattgc attttatctg 660 atagaatggt aggcaccata aataatcagg agattcanat ggagttagtt gaaatcgctt 720 caatagttgc cgaggatcga ttcctgcagc ccggggatca ctagttctag agcggcgccc 780 acgcggtgga gctccagctt tgtcccttag tgaggttaat ttcagctt 828 <210> 21 <211> 736 <212> DNA <213> Bacteriophage YS01 <400> 21 ccttcgcaac tatttgaagc gatttcaact aactccattt gaatctcctg attatttatg 60 gtgcctacca ttctatcaga taaaatgcaa taaaaaacct cccgaaggag gcttatttaa 120 ttaacgccaa acttatttct gtttaacctg aacctctacc agaatgccat caacactatc 180 gtcttgagtt ggcattgcgc cggctgctgt tgctgctgcg atatagcctc cagcaaaagt 240 ccaaccagta gtgaaagctg ccgcaccagc gactgctgca tcaacaacca aacctgcggc 300 attgataaat actggtttac caacggctcg gtttgcggcg gtaattgctg ctgctgccct 360 tacccagata cgaccagcag tcattacgtt aacagcctcc tgagctttgg ctgtgccatc 420 tggagtctca taatgagaac gaacaacaac accgtaagcg ctaccttctt catttactgc 480 ggtaacaacc ttgtgaccat caatctcagg agcggtagtt acggaaaccg ccacaccaac 540 agggattgca tcagagccac caactacanc agcgccatcg atattataca ggggagtatc 600 agctaccata cctgccagtg ctacagaacg aaatgcccat tatctatctc cctacttttt 660 aattttcttc anngcagcgc gcgggtcaag gtcaatgatg accctcgagg gggggcccgg 720 gtaccaattt cgcctt 736 <210> 22 <211> 783 <212> DNA <213> Bacteriophage YS01 <400> 22 gtagaactaa gtaatgatga ggttattgag caaatcaaga aagatggtaa atgggttaat 60 cttgaagagc acggaaaata cataaagaca aaaacgcttt acgtgacgcg cgttaaccac 120 aaatctggcg ttaagaagga tttcaaagaa ggaaagcgct atcaggtcaa tatgcacagc 180 ggcatgggcc aatcagctgg ttacatttac gatgaagatg gtaatgcatg gcagctatat 240 cgcagtgaag atgttggatt ccacacattg tgtggcacat attactttga ggcacaatac 300 aaatgattca tgaactaaag atactaccaa tccatttcga gccagttgag gatggcctta 360 agttagctga acttagaaaa aatgatagaa atttttccat aggagacaag ctaatactta 420 aagagttttc agatggagaa tacacaggaa gggttgttat aaaggaagtt ttacatatag 480 cagatgtttc cgcttatctt catggatacg ttttgctttc aatgcattaa tatcatttta 540 gacaatcaga tttaccatag gtgcgttaac attagcgcga caactaagga gttttttaat 600 gtttgatttc aacgaagata agctatctgt tgaacaagtt atggncattg ccgcgaaacg 660 acaccttcac cgcttcgcgt ggctattaat gctaaatggt atcgacaatc tcaatctttc 720 tggaaacacc agtcgagatt gacgctggta aatgacagta cccagttatc tcgctaggta 780 cga 783 <210> 23 <211> 739 <212> DNA <213> Bacteriophage YS01 <400> 23 atctaagtat ttcgatataa aagatgtaca gagatgttgg ctgctgagtt ccgtgatact 60 caacagtaaa gcgcccaagc atagcggcag acacgcaacc aagaaagtgc atgtaagcgg 120 aagactcggg aaactgaact gaacgagcaa tattagctga gagcttacca actacatcaa 180 tgtcgttacc tagcgagata actgggtact tgtcattacc agcgtcaatc tcgactggtg 240 ttttccagaa agattgagat tgtcgataac cattagcatt aatagccacg cgaagcggtg 300 gaaggttgtc gttcgcggca attgccataa cttgttcaac agatagctta tcttcgttga 360 aatcaaacat taaaaaactc cttagttgtc gcgctaatgt taacgcacct atggtaaatc 420 tgattgtcta aaatgatatt aatgcattga aagcaaaacg tatccatgaa gataagcgga 480 aacatctgct atatgtaaaa cttcctttat aacaaccctt cctgtgtatt ctccatctga 540 aaactcttta agtattagct tgtctcctat ggaaaaattt ctatcatttt ttctaagttc 600 agctaacttt agggcatcct caactggctc gaaatggnat ggtagtatct ttagttcatg 660 aatcatttgt aatgtgcctc caagtaatat ggggcacacc atgtgtggaa tcccacatcc 720 tccctgcgat ataactgcc 739 <210> 24 <211> 470 <212> DNA <213> Bacteriophage YS01 <400> 24 ccttgattcc agcattccag ccttacgaca gtattattct ggtaggtgat ttacctgtca 60 ggtgtttaat ggtatccggc acagacctaa gtatcactga tatcacccgc atgattgatg 120 cgatcgagac tagcacccaa ggcttgcata ctgccactga tgtagcaatc ccatacgaag 180 atgagatatt ggatgacctt attttcttca atttgctggt tgggcgcgaa ttcacttgtg 240 atggagataa acgcgaagct ggttcaaatt acgaggatgg tgaataatgt ttggattaaa 300 cgacgcatac tttaatgcag ttaagcgcca ggctgataag ctaaacgacg aatacggcaa 360 gctgggtgcg aagaagaaag atgacaaagt tgtggcggca ctgattactc aggtttgggg 420 tccagtttct acgctaatcg cgagggataa gtttgtgtgg atagcgggat 470 <210> 25 <211> 735 <212> DNA <213> Bacteriophage YS01 <400> 25 atcccgctat ccacacaaac ttatccctcg cgattagcgt agaaactgga ccccaaacct 60 gagtaatcag tgccgccaca actttgtcat ctttcttctt cgcacccagc ttgccgtatt 120 cgtcgtttag cttatcagcc tggcgcttaa ctgcattaaa gtatgcgtcg tttaatccaa 180 acattattca ccatcctcgt aatttgaacc agcttcgcgt ttatctccat cacaagtgaa 240 ttcgcgccca accagcaaat tgaagaaaat aaggtcatcc aatatctcat cttcgtatgg 300 gattgctaca tcagtggcag tatgcaagcc ttgggtgcta gtctcgatcg catcaatcat 360 gcgggtgata tcagtgatac ttaggtctgt gccggatacc attaaacacc tgacaggtaa 420 atcacctacc agaataatac tgtcgtaagg ctggaatgct ggaatcaagg atcaagctta 480 tcgataccgt cgacctcgag gggggggccc ggtacccaat tcgccctata gtgagtcgta 540 ttacaattca ctgggcgtcg ttttacaacg tcgtgactgg gaaaaaccct ggcgttacca 600 acttaatcgg cttgcagcac atcccccttt tcgcagctgg cgtaataccg aagaaggccg 660 caacgatcgc ccttcccaaa catttgccaa cctgaatgcc aaatgggaag cgccctggaa 720 acgcgcatta acccg 735 <210> 26 <211> 724 <212> DNA <213> Bacteriophage YS01 <400> 26 atcattgacc ttgacccgcg cgcttgcctg aagaaaatta aaaagtaagg agatagataa 60 atggcaattc gttctgtagc actggcaggt atggtagctg atacttccct gtataatatc 120 gatggcgctt gtgtagttgg tggctctgat gcaatccctg ttggtgtggc ggtttccgta 180 actaccgctc ctgagattga tggtcacaag gttgttaccg cagtaaatga agaaggtagc 240 gcttacggtg ttgttgttcg ttctcattat gagactccag atggcacagg caaagctcag 300 gaggctgtta acgtaatgac tgctggtcgt atctgggtaa gggcagcagc agcaattacc 360 gccgcaaacc gagccgttgg taaaccagta tttatcaatg ccgcaggttt ggttgttgat 420 gcagcagtcg ctggtgcggc agctttcact actggttgga cttttgctgg aggctatatc 480 gcagcagcaa cagcagccgg cgcaatgcca actcaagacg atagtgttga tggcattctg 540 gtagaggttc aggttaaaca gaaataagtt tggcgttaat taaataagcc tccttcggga 600 ggttttttat tgcattttat ctgatagaat ggtaggcacc atanataatc aggagattca 660 aatggagtta gttgaaatcg cttcaaatag ttgcgaaaga tcttttatat cgaaatactt 720 agat 724 <210> 27 <211> 727 <212> DNA <213> Bacteriophage YS01 <400> 27 atctaagtat ttcgatataa aagatccttc gcaactattt gaagcgattt caactaactc 60 catttgaatc tcctgattat ttatggtgcc taccattcta tcagataaaa tgcaataaaa 120 aacctcccga aggaggctta tttaattaac gccaaactta tttctgttta acctgaacct 180 ctaccagaat gccatcaaca ctatcgtctt gagttggcat tgcgccggct gctgttgctg 240 ctgcgatata gcctccagca aaagtccaac cagtagtgaa agctgccgca ccagcgactg 300 ctgcatcaac aaccaaacct gcggcattga taaatactgg tttaccaacg gctcggtttg 360 cggcggtaat tgctgctgct gcccttaccc agatacgacc agcagtcatt acgttaacag 420 cctcctgagc tttggctgtg ccatctggag tctcataatg agaacgaaac acaacaccgt 480 aagcgctacc ttcttcattt actgcggtaa caaccttgtg accatncatc tcangagcgg 540 tagttacgga aaccgccaca ccaacaggga attgcatcag aaccaccaac tacacaaggc 600 gcatcgatat tatacangga agtatcagct accataactg ccagtgctac gaaacaaatg 660 gcctttatct atctccttac cttttaaatt tcctcaagga agcgcgcggg tcaaggtcaa 720 tgatatc 727 <210> 28 <211> 702 <212> DNA <213> Bacteriophage YS01 <400> 28 atcggaactt cccttgctgg tggcatccca atctggctca ttgcaattca gcagggcggt 60 cagattaagg atagttttgg tggcattgct aataccgcaa ggtttgcatt agcagcaatt 120 aacccattaa cagcagttgt tgggactttg ggtttagctc ttggcgctct tggtgttgca 180 gcttaccagg cagacaagaa tactcgagga ctagcatcta cgataacgtt ggttggcaac 240 tcaactatca cgtctacagc gcagataaat caggttgttg atggaattga gagatcaaca 300 ctagctactc gcggtcttat tcaggagata gcgaacagcc ttgtagcaaa cggcaaccta 360 acgcttacgc agatagataa gataactaaa gcaactgctc agtgggcaac tgtaacatca 420 acggattcaa agacaatact tgggtacttt gatcaaatca caaaagaccc gatacaaggc 480 ttgattgcgc ttgataagca atttaacttc cttcagtccg ggcagcttaa atacattgag 540 agcatcagga agacggctgg agaaacagag gcagtaaccg cagctactga gttattcgct 600 aacgttatgg aaaacaggct tgcggacgtt gcgctaagcc tcaatcctct tngaagtgca 660 tggaatagct tcaggaagtt tgtgagtgag atctnggatg at 702 <210> 29 <211> 693 <212> DNA <213> Bacteriophage YS01 <400> 29 atcatcccag atctcactca caaacttcct gaagctattc catgcacttt caagaggatt 60 gaggcttagc gcaacgtccg caagcctgtt ttccataacg ttagcgaata actcagtagc 120 tgcggttact gcctctgttt ctccagccgt cttcctgatg ctctcaatgt atttaagctg 180 cccggactga aggaagttaa attgcttatc aagcgcaatc aagccttgta tcgggtcttt 240 tgtgatttga tcaaagtacc caagtattgt ctttgaatcc gttgatgtta cagttgccca 300 ctgagcagtt gctttagtta tcttatctat ctgcgtaagc gttaggttgc cgtttgctac 360 aaggctgttc gctatctcct gaataagacc gcgagtagct agtgttgatc tctcaattcc 420 atcaacaacc tgatttatct gcgctgtaga cgtgatagtt gagttgccaa ccaacgttat 480 cgtagatgct agtcctcgag tattcttgtc tgcctggtaa gctgncacac caagagcgcc 540 aagagctaaa ccccaagtcc aacactgctg ttaatggngt attgctgcta tgcaaccttt 600 gcgtatagca atgcacaaaa ctatcctaat ctgaccgcct gctgaatgca tgagcagatt 660 ggatgccaca gcaaggaagt ccgattcagc tat 693 <210> 30 <211> 856 <212> DNA <213> Bacteriophage YS01 <400> 30 nnnggggcgt cagggaggct gcaggcgata agttgggtaa cgccangggt tttcccagtc 60 acgacgttgt aaaacgacgg ttcgtgaatt cgagctcggt acccatccta cttggtgagt 120 ttgcaatagc ggcaaccaag acacttcagt caatacctgg acttggcgat ttaggtaata 180 aagcaatcgc tgagcagcaa aagattgtcg atgctgctaa gaagcaaaac attgagttag 240 cgaaaagcat agctgagcga gatgctagaa tcaggaaggg tgaacttggt tatgttgagg 300 gcaacaataa taccaacctg agcggcaata acgcatcaag cgataaggag tttgcagcaa 360 ggaagaaggc gcttgcggat gagtatgatg cgataaagaa agcgaggagt gagaggagca 420 aggcagtaaa agaagaaagg gatttaactt tatcttatga atctggtgtt cttgctcttc 480 aggctcagct gaaggtttta caagagcaca ggacgataaa cgatgtaata agcaacgaac 540 gcaagcagct attccaggaa gaagctaagt ttgcaatact tgagcaaagg aaagcggatg 600 gcacgcttac aaaatcacag gctaagctac ttgcgcagaa gaatatcatt cttgaacaag 660 ctaaacagaa ggctgaactt ggtgaccaga ttgtacttca ggagaggtca aacaagttac 720 ttgatgacaa cccttagaag actgtgcaga ttaagaatga ggncaataat ggttcgcttg 780 ctgctaattt atctgacagg nagaagcaga agagcaaaga gcttcaggca cttcaatcca 840 tcagataaac aaggtg 856 <210> 31 <211> 869 <212> DNA <213> Bacteriophage YS01 <400> 31 nnnnnnaagt atgtgcgatc ttatgcagct ggcacgacag gtttcccgac tggaaagcgg 60 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac 120 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag 180 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaggat 240 ccccccttgc tccatactct tcgccacgga atgtaattgt cttggctggt agtgactgac 300 cagatgcctg agcttgcatg atttcctcgg gcgtgtatga gatgttctcc ccatgaaacc 360 tgtaaaccct tgcgccgaac ttactgccgt caacctcaat aagcgtaacc acctcgccag 420 ggaaaagtga ctgaagctgg ttctcaaact ttacacttgc catattctct ccaataaaaa 480 aggccctcaa tgggccttag tttaactcat tgacgtgaaa acttcagtaa attgcatgct 540 aacagtctga acttctttcg tgcttggcga gacgctaatg ctatcctgct gaataaaaaa 600 cagctttagt tcgccctgcg gtgtagtcca tgcgaaaggt tttacgatgt gatcaaagca 660 gaagttcatt accgcctccc aatcacgacc gacgtagcta acagtatatg tcctctntgt 720 agtcctgtat ccaccagttc cgcgctgctg ataaccattt cgcaatgaga caaccctaac 780 attgttggcg ttagtaaaat gagaagcctc cattgagaac attttaacac caattaaaag 840 tttgcaatgc cattttaaat cctttaggg 869 <210> 32 <211> 850 <212> DNA <213> Bacteriophage YS01 <400> 32 nnggggcgtc aggaggctca ggcgattagt tgggtaacgc cangggtttt cccagtcacg 60 acgttgtaaa acgacggcca gtgaattcga gctcggtacc cgtaaacaag tttatgataa 120 cgactgtctt gatcaagcac gtgcttacta gttggagtta cagcctcaat tccatctgag 180 tgaaggaaat caccaccaat cagtaaaact gccttagatg aatttggagc tttagcgacc 240 gcataatcaa aatattggtt aagcactctc tcagcaattt cggtgctgta gttttccccg 300 cactcagcat ggtgagccat tgcgccaatg tgcatatcaa atacagggta gaggctcaat 360 gtgtcttcgt agcttgattc actgagcggc tttggagtct cgcgagggac ttcatctgta 420 agagcctgaa caatctgctg aagcatctct tctagcttct cggcatcaat tgcagtctta 480 acccagcgca gcttctcatt gccttccgca tcaatcatgg ttgaagtgcc cttgactaaa 540 tatccatccg gcactagctt cttgatacct tcgttcccat gcccaatacc actcttagcc 600 aacttgctgc gacgaagttc aacgttgcga atgttcatac cgtacttctc tgcaatggcc 660 ttattagtca tgccattttt caactcttcg ataagctgct ctgtggaaat ctttgctact 720 gccattttta agtccttagt tcatcacgat aaaatataag atagtaaagc taaacagtgg 780 tactaatgct actagggctg cgtacttcat acaacctctt tcaccagtaa tcaattaaat 840 ccgcactaat 850 <210> 33 <211> 855 <212> DNA <213> Bacteriophage YS01 <400> 33 nnnnnnnggc agtggcgatc ttatgcagct ggcacgacag ggtttcccga ctggaaagcg 60 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta 120 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca 180 ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga 240 tccccatcac tgcgaagttg ccctgtcggt tgttaattcg cgcaagacct tcaggcgtag 300 tatcggatac cgggaacaca atatcacaca gcttctcaag tttttcctcg aggtcttctt 360 tttcttcaaa cagtgaagcc atctcagaag gtgacttctc ctgtttcatc tctttagata 420 gagccgacag tttagccata atctttttgc gatcacgttt acgcacttca ttcaaccgct 480 cagtctcagc aatcattggg gcaatcgaca gggagttgat agccgatttg ccagttgatg 540 gaggctggct agttactacc ctcgcaagta tgtcagggtt aagcctggtg tcacgataaa 600 cacgaatgag ttcctganag atcagggatt caaatttaag ataacgaagg gttagttatg 660 ttgggattag atttttcacc ctgagcatta tgatttagtt catggcagta gtggtgctaa 720 atttagagtg atacctatta ctgactggtt tccacctgac tatggggaat gtaatgcgaa 780 gaccaaagat ggtaagtggg gggcaaaatt accactcttc agctttgcgg tatctctggt 840 tgacaaacct gaacg 855 <210> 34 <211> 853 <212> DNA <213> Bacteriophage YS01 <400> 34 nnnnggggcg tcagggaggc tgcaggcgat taagttgggt aacgcncagg gttttcccag 60 tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg gtacccatct gacgaagcaa 120 caagcgtaaa ctcactgctt ggcatgacgt atggcgatgg ctctaagaaa acgaacagcg 180 aactcgtcct taaagcctgg gatgctggta acgtgtccat tgcgcgtgcg aacgcagaaa 240 acaatatgag cttcgtagct atgggatgta tctctgtaat tgcccaggat gaaacaatta 300 acgccatcat gaatgccggt gctcgtggta ttggtgtgtc ggaacgtttt cttctggtgc 360 gtgaaaagtc atttcttggc gatcgcgtgt ttgttgatga gaatggtaat tcaacatatg 420 aggctattga tggtgggcta aaggctgatt acttccgact cgttcatgag attatgaatg 480 aagtagaagt gaaactaact gtaagtgctt cttcaatgcg ttacctgaac cgtgcgcgcc 540 aggaaatgga gccgcatctt ggtgacggag gtaagtattc tcacacaatg ttgcgcggtg 600 ctctaggtaa gtttgataag caagcaatcc gtatcgcttc tgttctgcac gttattcgta 660 actggttcgc accagcaggc tccagccctc ataaatccag agagattgaa ctggagacca 720 tgcangaagc gggtatgaat tttcacgaac taagtaaaac ttatttgtca tctgccaacg 780 ctgctggtca cgctggtgac catgctgaga tgaataaact gattgatggg atctcctaga 840 gtccacctgc agg 853 <210> 35 <211> 856 <212> DNA <213> Bacteriophage YS01 <400> 35 nttttgcggc aatgcgattt tatgcagctg gcacgacagg gtttcccgac tggaaagcgg 60 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac 120 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag 180 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaggat 240 cccatcaatc agcttattca tctcagcatt gtcaccagcg tgaccagcag cgtttgcaga 300 tgacaaataa gttttactta gttcgttgaa aatcataacc gcttcctgca ttgtctccag 360 ttcaatctct ctggatttat gagggcttga gcctgctggt gcgaaccagt tacgaataac 420 gtgcagaaca gaagcgatac ggattgcttg cttatcaaac ttacctagag caccgcgcaa 480 cattgtgtga gaatacttac ctccgtcacc aagatgcggc tccatttcct ggcgcgcacg 540 gttcaggtaa cgcattgaag aagcacttac agttagtttc acttctactt cattcataat 600 ctcatgaacg agtcggaagt aatcagcctt tagcccacca tcaatagcct catatgttga 660 attaccattc tcatcaacaa acacgcgatc gccaagaaat gacttttcac gcaccagaag 720 aaaacgttcc gacacaccaa taccacgagc accggcattc atgatggcct taaatgtttc 780 atcctgggca attacagaaa tacatcccat agctacgaag ctcatattgg tttctgcggt 840 cgcacgcgca atggac 856 <210> 36 <211> 855 <212> DNA <213> Bacteriophage YS01 <400> 36 nnnggggcgt cagggaggct gcaggcgata agttgggtaa cgccangggt tttcccagtc 60 acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt accatcagca aaaggacact 120 tggggcgctt aacttaataa ctgacgttgt tgctggtacg attgagcaga taaggtcgtt 180 aataaactcc ggtgatatcc cgctatccac acaaacttat ccctcgcgat tagcgtagaa 240 actggacccc aaacctgagt aatcagtgcc gccacaactt tgtcatcttt cttcttcgca 300 cccagcttgc cgtattcgtc gtttagctta tcagcctggc gcttaactgc attaaagtat 360 gcgtcgttta atccaaacat tattcaccat cctcgtaatt tgaaccagct tcgcgtttat 420 ctccatcaca agtgaattcg cgcccaacca gcaaattgaa gaaaataagg tcatccaata 480 tctcatcttc gtatgggatt gctacgggga tcctctagag tcgacctgca ggcatgcaag 540 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 600 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 660 actcacatta attgcgttgc gctcactgcc cgctttccag tcnggaaacc tgtcgtgcca 720 gctgcattaa tgaaatcgnc aacgcgaatt nccgacagta agacgggtaa gcctgntgat 780 gataccgctg ccttactggg tgcattagcc agtctgaatg acctgtcacn ggataatccg 840 aagtggtcag actgg 855 <210> 37 <211> 845 <212> DNA <213> Bacteriophage YS01 <400> 37 nnngggccgg tgttgcattt tatgcagctg gccgacaggt ttcccgactg gaaagcgggc 60 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac 120 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga 180 aacagctatg accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc 240 ccgtagcaat cccatacgaa gatgagatat tggatgacct tattttcttc aatttgctgg 300 ttgggcgcga attcacttgt gatggagata aacgcgaagc tggttcaaat tacgaggatg 360 gtgaataatg tttggattaa acgacgcata ctttaatgca gttaagcgcc aggctgataa 420 gctaaacgac gaatacggca agctgggtgc gaagaagaaa gatgacaaag ttgtggcggc 480 actgattact caggtttggg gtccagtttc tacgctaatc gcgagggata agtttgtgtg 540 gatagcggga tatcaccgga gtttattaac gaccttatct gctcaatcgt accagcaaca 600 acgtcagtta ttaagttaag cgccccaagt gtccttttgc tgatggtacc gagctcgaat 660 tcactggccg tcgttttaca acgtcgtgac tggaaaaacc tggcgttacc caacttaatc 720 gccttgcagc acatcccccc ttcgncagct ggcgaatagc gaaaaggccc gcacgatcgc 780 ctttccaaca gtgcgcagcc tgatgccaat gcggctgatg cgtatttctc ttaccatctg 840 ggcga 845 <210> 38 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> M13 forward primer <400> 38 gtaaaacgac ggccag 16 <210> 39 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> M13 reverse primer <400> 39 caggaaacag ctatgac 17  

Claims (8)

E. coli (Escherichia coli) specific infection and killing ability, the genome comprising a nucleic acid molecule represented by a tail protein having the amino acid sequence of SEQ ID NO: 1 and the base sequence consisting of SEQ ID NO: 2 to SEQ ID NO: 37 Siphoviridae family, characterized in that it has a bacteriophage belonging to the genus T1-like phage (T1-like phage genus). The bacteriophage according to claim 1, wherein the bacteriophage is bacteriophage YSO 1 having accession number KCCM10947. i) seedling the microorganisms that are not E. coli, and ii) inoculating the cultured microorganism with the bacteriophage having E. coli specific infection and killing ability in the culture medium, wherein the bacteriophage has a amino acid sequence of SEQ ID NO. And a genome comprising a nucleic acid molecule represented by a nucleotide sequence consisting of SEQ ID NO: 2 to SEQ ID NO: 37 in the Siphoviridae family, T1-like phage genus A method for preventing or controlling contamination of E. coli in fermentation by culturing a microorganism which is a belonging bacteriophage. delete The method according to claim 3, wherein the bacteriophage is bacteriophage YSO1 having accession number KCCM10947. The method according to claim 3 or 5, wherein the microorganism other than E. coli is Corynebacterium having L-amino acid production ability. The method of claim 3 or 5, wherein the bacteriophage is inoculated at a concentration of 10 ⁴ pfu to 10 ⁶ pfu per 1 L of the culture medium in step ii). Or how to control. The method of claim 3 or 5, wherein the temperature of the culture medium inoculating the bacteriophage in step ii) is 30 ℃ to 33 ℃ prevent or control the contamination of E. coli in fermentation by the culture of microorganisms How to.

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