Akkermann mucophilic bacteriophage AkkZT003P and application thereof

Abstract

The invention relates to ackermann mucophilic bacteriophage AkkZT003P and application thereof, and belongs to the technical field of microorganisms. The phage provided by the invention is Akkermansia muciniphila phage AkkZT003P, the phage is preserved in the Guangdong Microbial Culture Collection Center on January 6, 2025, and the preservation number of the phage is GDMCC (China General Microbiological Culture Collection Center) NO: 65724-B1. According to the invention, Ackermann mucophilum and Ackermann mucophilum bacteriophage are successively separated from excrement through in-vitro experiments, and animal experiments prove that the bacteriophage can crack Ackermann mucophilum to maintain the integrity of mucus. The effect of the bacteriophage on Ackerman muciniphile can lead to increase of downstream symbiotic bacteria streptococcus thermophilus, so that the intestinal barrier is further protected, the intestinal inflammation is improved, and the intestinal health is recovered.

Description

Technical Field

[0001] The present invention relates to the technical field of microorganisms, and in particular to an Akkermansia muciniphila phage AkkZT003P and its application. Background Art

[0002] Akkermansia muciniphila (A. muciniphila) is one of the common symbiotic bacteria in the human intestine. It is a Gram-negative obligate anaerobe and is known for its ability to degrade mucin. Mucin is the main component of the mucus layer in the human gastrointestinal tract and is crucial for maintaining the intestinal barrier function. Although it is considered a beneficial bacterium for colorectal cancer and metabolic diseases, it is worth noting that due to the nature of A. muciniphila's foraging on intestinal mucus, in the case of mucosal barrier defects, such as in the case of inflammatory bowel disease (IBD), it will exacerbate intestinal inflammation. Some existing studies have shown that an increase in the abundance of A. muciniphila in IL10 - / - mice can promote intestinal inflammation, or exacerbate Salmonella typhimurium-induced intestinal inflammation by disrupting host mucus homeostasis.

[0003] The human digestive system contains diverse microorganisms, such as bacteria, archaea, fungi, and phages. Among them, intestinal phages are key members of a healthy microbial community, and their ratio to bacteria ranges from 1:1 to 10:1. Phages, with their advantages of safety, specificity, no residue, anti-drug resistance, and rapid R & D, can effectively target specific bacteria without affecting the normal flora. Therefore, using phages to precisely target pathogenic bacteria in the intestine to promote the growth of beneficial bacteria in the intestine is gradually becoming an effective treatment strategy for promoting intestinal health.

[0004] Currently, there has been no report on isolating phages of A. muciniphila from feces. Although current studies have shown that excessive colonization of A. muciniphila in the intestine may exacerbate intestinal inflammation, the research reports on how to control the excessive colonization of A. muciniphila by treatment means to alleviate intestinal inflammation are relatively limited. Summary of the Invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an Akkermansia muciniphila phage AkkZT003P that lyses Akkermansia muciniphila and has an intestinal protection function and its application.

[0006] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0007] In a first aspect, the present invention provides a phage, characterized in that the phage is Akkermansia muciniphila phage AkkZT003P, which was deposited at the Guangdong Microbial Culture Collection Center on January 6, 2025, and its deposit number is: GDMCC NO: 65724 - B1.

[0008] In the present invention, Akkermansia muciniphila phage AkkZT003P promotes the growth of Streptococcus thermophilus by lysing Akkermansia muciniphila, thereby improving intestinal inflammation, and is a gut - protective phage.

[0009] In a second aspect, the present invention provides the application of the phage in the preparation of a drug for preventing and / or treating enteritis.

[0010] Furthermore, the drug is a drug that inhibits Akkermansia muciniphila.

[0011] Furthermore, the drug is a drug that promotes the growth of Streptococcus thermophilus.

[0012] Even further, the drug is a drug that reduces the expression of intestinal pro - inflammatory cytokines.

[0013] In a specific embodiment of the present invention, the pro - inflammatory cytokines include at least one of interleukin - 1, interleukin - 6, and tumor necrosis factor α.

[0014] Furthermore, the drug is a drug that protects the intestinal barrier.

[0015] Even further, the drug is a drug that increases the expression of intestinal barrier factors.

[0016] In a specific embodiment of the present invention, the barrier factor includes human mucin 2.

[0017] In a third aspect, the present invention provides a drug for preventing and / or treating enteritis, and the drug contains the phage described above.

[0018] Furthermore, the drug further contains a pharmaceutically acceptable carrier and / or excipient.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0020] In this invention, Akkermansia muciniphila and Akkermansia muciniphila phage AkkZT003P were successively isolated from feces through in vitro experiments. Through animal experiments, it was verified that the phage could lyse Akkermansia muciniphila to maintain the integrity of mucus, and the effect of the phage on Akkermansia muciniphila could downstream lead to the growth of symbiotic bacterium Streptococcus thermophilus, further protecting the intestinal barrier, improving intestinal inflammation, and restoring intestinal health. Brief Description of the Drawings

[0021] Figure 1 To isolate Akkermansia muciniphila ZT003. Among them, a is the growth morphology; b is the whole genome sequencing.

[0022] Figure 2 To isolate Akkermansia muciniphila phage AkkZT003P and conduct biological characteristic characterization. Among them, a is the phage isolation flow chart; b is the transmission electron micrograph of the phage; c is the analysis of the phage whole genome sequence; d is the lysis situation of the phage on Akkermansia muciniphila.

[0023] Figure 3 To verify through in vitro experiments that Akkermansia muciniphila phage AkkZT003P lyses Akkermansia muciniphila, thereby promoting the growth of Streptococcus thermophilus. Among them, a is the in vitro experiment flow chart; b is the influence of the phage on the growth of Akkermansia muciniphila and Streptococcus thermophilus.

[0024] Figure 4 To verify through animal experiments that Akkermansia muciniphila phage AkkZT003P lyses Akkermansia muciniphila, promotes the growth of Streptococcus thermophilus, and achieves an anti-inflammatory effect. Among them, a is the animal experiment flow chart; b is the change in the body weight of mice; c is the change in the colon length of mice; d is the histopathological result of the intestinal tissue of mice; e is the gene expression level of pro-inflammatory cytokines and barrier factors in the intestinal tissue of mice. Detailed Embodiments

[0025] To better illustrate the purpose, technical solution, and advantages of this invention, the following will further illustrate this invention in combination with specific embodiments. Other materials, reagents, etc. used in the embodiments can be obtained from commercial channels without special instructions.

[0026] Example 1 Isolation of Akkermansia muciniphila ZT003

[0027] I. Experimental Methods

[0028] 1. Inoculate 100 mg of fresh fecal samples into Brain Heart Infusion Broth (BHI) selective medium, add 0.2% (m / v) mucin and antibiotics (vancomycin 6 μg / mL, gentamicin 10 μg / mL, and kanamycin 12 μg / mL), and culture under anaerobic conditions at 37°C for 48 h to obtain a bacterial suspension.

[0029] 2. Spread the bacterial suspension from step 1 onto the BHI agar selective medium supplemented with mucin and antibiotics in step 1, and select single colonies for PCR identification.

[0030] (1) Experimental procedure: Extract the bacterial liquid DNA of single colonies as a template, design primers according to the specific gene sequence (outer membrane protein Amuc-1100) of the target strain Akkermansia muciniphila (primers: 5’-TGGCGAACAATCCGGAAGAA-3’; 5’-GGGGCAT CATCCGTTCGTTA-3’), use a commercial PCR amplification kit (brand: TAKARA; catalog number: R010A), and perform PCR amplification according to the instructions. Set the program as follows: denaturation (94 - 95°C, 30 s), annealing (65°C according to the Tm value of the primers, 30 s), extension (72°C, 30 s - 1 min), number of cycles (38 cycles), and final extension (72°C, 5 - 10 min) to obtain PCR products.

[0031] (2) Result analysis: Electrophorese the PCR products in 1 - 2% (w / v) agarose gel, stain with GelRed non-toxic nucleic acid dye (a fluorescent nucleic acid gel staining reagent), and observe specific electrophoresis bands. If a band of the same size as the specific gene of the target strain (183 bp) appears, the strain can be preliminarily identified as the target strain.

[0032] 3. Further identify the strain in step 2 by whole-genome sequencing.

[0033] II. Experimental Results

[0034] As Figure 1 shown in Figure 1 a and b, the growth morphology and whole-genome sequencing results of the strain isolated in this example. The strain isolated in this example is Akkermansia muciniphila, named Akkermansia muciniphila ZT003.

[0035] Example 2 Isolation and Biological Characterization of Akkermansia muciniphila phage AkkZT003P

[0036] I. Experimental Methods

[0037] 1. Isolation of Phage

[0038] The flow chart for phage isolation is shown in Figure 2 a. Collect an appropriate amount of fecal samples, suspend them in SM buffer (sodium chloride magnesium sulfate buffer), stir evenly, place them at 4°C overnight, centrifuge at 10000 rpm for 10 min, and filter through a 0.22 μm filter to obtain fecal supernatant containing phage particles.

[0039] Enrichment: To improve the isolation effect of phage, mix 100 μL of fecal supernatant, 100 μL of Akkermansia muciniphila ZT003 isolated in Example 1 (OD600 = 0.6), and 5 mL of BHI broth medium, incubate anaerobically at 37°C for 24 h, centrifuge at 10000 rpm, and filter through a 0.22 μm filter. Repeat this enrichment process 3 - 5 times to obtain a high-titer phage lysate.

[0040] Isolate single phages from the phage lysate using the double-agar overlay method and perform further amplification and enrichment using the plate method.

[0041] Directly titrate 20 μL of phage onto the semi-agar layer of the plate medium mixed with 100 μL of Akkermansia muciniphila ZT003 (OD600 = 0.6), incubate anaerobically at 37°C overnight to form large plaques. Immerse the plate medium containing plaques in 4 mL of SM buffer and place it at 4°C for more than 4 h to obtain a lysate. Centrifuge the lysate at 5500×g for 10 min and filter it aseptically through a 0.22 μm filter membrane to obtain the final high-titer phage, achieving the amplification and enrichment of the phage.

[0042] 2. Whole-Genome Sequencing and Characterization of Phage

[0043] Prepare high-titer phage using the solid titration method for whole-genome sequencing of the phage.

[0044] Mix 100 μL of the Akkermansia muciniphila ZT003 (OD600 = 0.6) isolated in Example 1 with 0.7% (w / v) of upper soft agar, and pour it onto 1.5% (w / v) lower agar to obtain a plate medium containing Akkermansia muciniphila.

[0045] Take 10 μL of the phage isolated in this example, drop it onto the plate medium containing Akkermansia muciniphila and culture it overnight to form large plaques. Take the plaque and soak it in 4 mL of SM buffer, culture it at 37 °C for 24 h, collect the lysate, centrifuge it at 5500×g for 10 min, and filter and sterilize it with a 0.22 μm filter to obtain a phage stock solution with a high titer. Take a transmission electron microscope (TEM) picture of the phage with a high titer, and extract the genomic DNA of the phage using the MinElute Virus Spin Kit (brand: QIAGEN; product number: 57704), and perform whole-genome sequencing.

[0046] 3. Detect the lysing ability of the phage against Akkermansia muciniphila.

[0047] Take 5 mL of BHI selective medium supplemented with 0.2% (m / v) mucin, add 50 μL of Akkermansia muciniphila ZT003 in the logarithmic growth phase, and add 100 μL of SM buffer or phage at a concentration of 10 4 PFU / mL to it respectively. Collect the bacterial liquid at 0, 6, 12, 18, 24, and 36 h to detect OD600 to monitor the lysing ability of the phage against Akkermansia muciniphila.

[0048] II. Experimental Results

[0049] 1. Transmission electron microscope (TEM) shows that the phage is a long-tailed ds-DNA phage, with an icosahedral head with a diameter of 50.66 nm and a tail of 132.58 nm ( Figure 2 b). Whole-genome sequence analysis shows that the phage has a circular genome with a full length of 31848 bp, containing an integrase gene ( Figure 2 c), and it is a phage of the genus Fernvirus. The whole-genome nucleotide sequence is shown in SEQ ID NO: 1.

[0050] 2. As shown in Figure 2 d, the phage can effectively lyse Akkermansia muciniphila.

[0051] The phage was named Akkermansia muciniphila phage AkkZT003P and deposited at the Guangdong Provincial Microbial Culture Collection Center on January 6, 2025. Its deposit number is GDMCC NO: 65724 - B1, and the deposit address is on the 5th floor of Building 59, No. 100 compound, Xianlie Middle Road, Guangzhou.

[0052] Example 3: In vitro experiment to verify that Akkermansia muciniphila phage AkkZT003P lyses Akkermansia muciniphila, thereby promoting the growth of Streptococcus thermophilus

[0053] I. Experimental method

[0054] As Figure 3 shown in a, to verify the competitive relationship among Akkermansia muciniphila phage AkkZT003P, Akkermansia muciniphila, and Streptococcus thermophilus in vitro, 50 μL of Streptococcus thermophilus CICC 6038 (Wuhan Huizao Biotechnology Co., Ltd., product number HZB196192) in the logarithmic growth phase and Akkermansia muciniphila ZT003 were simultaneously inoculated into 5 mL of culture medium, and 100 μL of Akkermansia muciniphila phage AkkZT003P (1×10 4 PFU / mL) or SM buffer was added respectively. Anaerobic incubation was carried out at 37 °C for 24 h, and qPCR was used to quantify the growth of Akkermansia muciniphila and Streptococcus thermophilus.

[0055] II. Experimental results

[0056] As Figure 3 shown in b, Akkermansia muciniphila phage AkkZT003P can lyse Akkermansia muciniphila in vitro and promote the growth of Streptococcus thermophilus.

[0057] Example 4: Animal experiment to verify that Akkermansia muciniphila phage AkkZT003P lyses Akkermansia muciniphila, promotes the growth of Streptococcus thermophilus, and achieves an anti - inflammatory effect

[0058] I. Experimental method

[0059] As Figure 1 shown in a, it is the experimental procedure for mice in vivo.

[0060] Grouping and treatment of mice: Male C57BL / 6J mice at 6 weeks of age were pretreated with a mixed antibiotic for 3 days by drinking water administration. The types of antibiotics in the mixed antibiotic were neomycin (brand: Aladdin; catalog number: N109017-25g), metronidazole (brand: Aladdin; catalog number: M109874-25g), ampicillin (brand: Aladdin; catalog number: A105483-25g), and vancomycin (brand: Aladdin; catalog number: V301569). Among them, the concentration of neomycin was 1 g / L, the concentration of metronidazole was 1 g / L, the concentration of ampicillin was 1 g / L, and the concentration of vancomycin was 0.5 g / L.

[0061] Mix Streptococcus thermophilus CICC 6038 at 1×10 8 CFU / mL and Akkermansia muciniphila ZT003 at 1×10 8 CFU / mL together, and gavaged 200 μL per mouse to allow the strains to colonize for 2 weeks. Then, 200 μL of SM buffer or the same volume of Akkermansia muciniphila phage AkkZT003P at a concentration of 10 6 PFU / mL was gavaged to the mice for 1 week. Before phage gavage, each mouse was gavaged with 100 μL of sodium bicarbonate to neutralize gastric acid, and phage gavage was performed 15 min later.

[0062] Establishment of a mouse colitis model: Mice were fed with 2.5% (w / v) dextran sulfate sodium (DSS) for 7 days, and the body weight changes of the mice were monitored during this period.

[0063] The results of mouse intestinal tissue pathology showed that intestinal tissues of mice in each group were collected for hematoxylin-eosin staining (H&E staining).

[0064] RT-qPCR was used to detect pro-inflammatory cytokines and barrier factors in mouse intestinal tissues. The pro-inflammatory cytokines were interleukin-1 (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α); the barrier factor was the gene expression level of human mucin 2 (Muc-2).

[0065] Mouse colon tissue samples were immediately immersed in RNAlater (brand: ThermoFisher; catalog number: AM7021) and stored at -20 °C until further processing. RNA was extracted from mouse colon tissue samples using the TransZol Up Plus RNA Kit (brand: TransGen Biotech; catalog number: ER501-01-V2). RNA was reverse transcribed into cDNA using HiScript III RT SuperMix for qPCR (brand: Vazyme; catalog number: R323-01). The expression levels of cytokines were detected by quantitative real-time PCR (qPCR) using qPCR SYBR Green MasterMix (without Rox) (brand: Yeasen; catalog number: 11201ES08) on a QuantStudio 7Flex real-time fluorescence quantitative PCR (Applied Biosystems). The primers were as follows:

[0066] β-actin: 5’-CGTTGACATCCGTAAAGACC-3’; 5’-AACAGTCCGCCTAGAAGCAC-3’.

[0067] IL-1β: 5’-GGATGAGGACATGAGCACCT-3’; 5’-AGCTCATATGGGTCCGACA-3’.

[0068] IL-6: 5’-TGGGAAATCGTGGAAATGAG-3’; 5’-CCAGTTTGGTAGCATCCATCA-3’.

[0069] TNF-α: 5’-GGTGCCTATGTCTCAGCCTCTT-3’; 5’-GCCATAGAACTGATGAGAGGGAG-3’.

[0070] Muc-2: 5’-ATGCCCACCTCCTCAAAGAC-3’; 5’-GTAGTTTCCGTTGGAACAGTGAA-3’.

[0071] II. Experimental Results

[0072] Mouse body weight detection: As Figure 4 shown in b, compared with the control group of mice (given SM buffer), after treatment with DSS, the experimental group of mice (given phage) showed a significantly smaller decrease in body weight, indicating that the intestinal inflammation of the experimental group of mice (given phage) was significantly alleviated;

[0073] Mouse colon intestine: AsFigure 4 As shown in c, compared with the control group mice, the experimental group mice showed a longer colon length.

[0074] The histopathological results of the mouse intestinal tissues showed that: as Figure 4 shown in d, compared with the control group mice, after DSS treatment, the experimental group mice showed a lesser degree of histopathological damage.

[0075] The RT-qPCR detection results showed that: as Figure 4 shown in e, after DSS treatment, the gene expression levels of pro-inflammatory cytokines in the intestinal tissues of the experimental group mice were significantly lower than those of the control group mice; the gene expression level of the barrier factor (Muc-2) in the intestinal tissues of the experimental group mice was significantly higher than that of the control group mice.

[0076] The above results indicate that the Akkermansia muciniphila phage AkkZT003P protects the intestinal barrier by lysing Akkermansia muciniphila, and at the same time promotes the growth of Streptococcus thermophilus, thus playing a protective role against intestinal inflammation, and it is an intestinal protective phage.

[0077] The complete genome nucleotide sequence (SEQ ID NO: 1) of the Akkermansia muciniphila phage AkkZT003P is as follows:

[0078] ATGGATGAAGAAAAAGATCCTCAAGCCATCTATAAGCAGAATGTCAGAGCTTGGCTTCGA

[0079] GACCGAGGTCTTTCTATGAAGTGGCTAGCTAGCTGGTTGGGGAAGTCAGAAGGAACAGTT

[0080] AAAAATTGGTTTTACGGAGGGAATCCTATTATTGAAGAGAATGTTGATAGTATTGATGCA

[0081] CTTCAGGTAAATTATCATAAAGACAATCGCTATCATTTTATAAATCCGGATTTCGATCCA

[0082] TCTCCAAAGGCGTATAATATTCCCAAGAAACAATTGATAACCAGACTCCTGTTTGCCCGG

[0083] AAGGGGCTCCCGCATGGGTCACTTATGGCGCTCTTTTTTCCACTCCGGTAGAAGTAGCGC

[0084] CTCAACTGTATCAGAGAAAAAAGAATGTTGATACTTCAATCTCAGCGACTATATGGACAG

[0085] GAGTTGGCACTATCTTTATGATTATCTCACTTCCTCTTATAGCCACACCTTGTTTTAGGA

[0086] AATACGGAGAACCATCTACTGTTATCTTGTCTCTTTTCCCTGCCATTCTGGGATACCTAT

[0087] TTATTGCTTATTGGGCTCATGTAGTAGGAGACCGCACTATTTGTCGTTACATAGGAGGTA

[0088] TTTTTTGTCTGGTAGTTAGCGGACTATGCTTCTTCAGCACATCAAGACTTTCGGGGTTTG

[0089] GACATTTGGGCATCGTATTTCTAATTTTTTCGATACTTTTTGCGATGCCTATTATCCGTT

[0090] TATTCATCAAAAAACGCAAAAAATAATCTTTATTAAATAAAAATGAATGTGAAAATATAG

[0091] TAAGAGAAAGCAAATAGAAAAAGTTCTTAATTTTATATTTATTCATTGAACTAGCAAATA

[0092] TTTATTATGAATTTCCATCTATCCTTTACAACGGGAAATAACCCCGAACAAGCCCCAAAG

[0093] CAACAAATAGATGAAACTGAAAAAAATCGCTGTGATTTTGAAAATACTCTTACTATAGAA

[0094] AACTATAAAATACAACAACAAGAAAGGCTTCGTGCGTTGGAAGTTTCATCACGAGCGCAA

[0095] GCCAATTGGGACAAAACAACTATAACAATTTCATCCTCCGCCATTGGTCTAATATATCTG

[0096] CTAAAAGGAGAAAATATAATACTTTCTACCATACATTTTCGTATATCACTGATTTTGTGG

[0097] GCCGCAGTTGTTTCTTGTGTAATGATTTCATTCCTTGCTGGCGCAACGCTTCATCGTGAG

[0098] TACGCTAACAAAATAGCTCATCAACAAAATGGAGATGAAAAACAAGCTGCCAAAAGTGAT

[0099] GAAGAAGCAAATAAAATGGATAAAATAGTAAAATATTGCAACAACGCAGCATTTCTTTTA

[0100] TTCTTGTTCGCATTAGCAAGTACTGTATATCTTTCCTTTGAAGCCATATCAAACAGCCAT

[0101] GCCAGAACAGCCAATCAACAACAGACAAACGAGAAAGTCGAACAAGGAATTAATCACACA

[0102] TAGTGACAGAAAACCAGTATATCTCCGTCCTATTTCCATGGATAAAATGAAACTCCCATC

[0103] TTCCCTTTTGTCCCAAGGAATTGTCTAAAGACAAGCCAAGACAAGAGAAATAAGAATATT

[0104] TTTCTTGGGTATCCTAAGAAAGACAATAGTTCTGCCTCAGCAAAAAATGACACAATGTGA

[0105] AAACAATCCTTGCTATTTTCACATTGTGCAACTATATTGTAGGCATGCCTCCTAAGATAC

[0106] GACAACTAGTTGCCGATCTTCGCAAGGCGGGCTTTGACAATACGGGCCGCGGCAAAGGTT

[0107] CCCACCGTCAGTATGAGCATCGGGCCTCCGGTACGGTAGCGACCATCTGCGGGCATGATG

[0108] GAGACGATGCCAAGCCCTATCTGGTCAAGCACGTCAGGGAGAAGATAGAAGAGGCAAAAA

[0109] GCAAGGGAGCGTAAGCTCCCTTGCAACCCCAAGAAAAACAACTATGAAAATGACTGAACA

[0110] GGACATGAAGGCGCTTGCCGGGAAATACAGCCGTTTCATCCGGTGGAGCGATGAAGATAA

[0111] TTGCTACATCGGTTCCCTGCCTGATTTTGAAAAGGACTGCACCCACGGGGAAACCCTGGA

[0112] AGAAGTCAACCGCAACCTTCAGGAAGTGGCGGAAATGTATATTGAACGTTATCTGGAAAA

[0113] GGGAATGGAACTCCCCCAGCCCCGGTCCATAGCTATTTCTCCGTCACCTTTCCGCGAAAC

[0114] AGGCAATGAAAAAAGCATTGCGCGGTTAAGGAAAAGCCAGGGAATGAGTCAGAAAGATTT

[0115] TGCCGCCGTGCTGGGAGTCAGTCCTTCCACGCTGGTCAAATGGGAACACGGCTTGCGGCG

[0116] CCCCTGCGGCCCTTCCGCAAAATTGCTTGACCTTATTGAAAAGCATCCTGAATTGATCAA

[0117] TTCCTGACGTTATTTTGCCCAGGCAACAAAAAAAGCCCCCTCAAATAGGGGGCGTGGATA

[0118] GAAACAAATAACCCTAAACAAGCTCCTGGGCCACGCTGAACGCATCGTCGCAGCGGTTCA

[0119] ACCAGCCACGCCCGAAGGTCGGGAACTGCTTGCACGAACGGTAAAACGCCTGACGCTTCT

[0120] CCTGCAGAGCGATAAGGAACACCGCTTCACCCGTGGCGGCCAGCTGGTCCTGTAGTTCCT

[0121] GCCGGGTCTTGGGGCCGACAATCCCGTCCACCGTGAGCCCGGCACCGTGAATATTCAGCG

[0122] CACGTTGCAGGATCTTCCCGGTATTCCTGCTCCCGGAATTGAAAAAATGGTCCCGCAGGA

[0123] TGAATTCAACGCCAGGAAAAGCGTCAGAACCCAGCCAGGAACGCACGGCGGCGGTATTAT

[0124] CCAGGACGTACTGAAGACAACCTTCCCAGGCGTCTTCACGTCTTCCGGCATCCAGCAGGG

[0125] CCTTCAACCTGTTAAACACGTCCGGTTCAATACCGTTGCAAATGCCGCAAATCTCCCACT

[0126] TGCCGCCCTTGTCGGCGGCGGGAAGGCGGGAAACGCGCAGGGAATCAGGCCCGGTGACGC

[0127] GGCTGTCTTCAAACCGGAGGATGGCCGCAGCCATCTTTCTTTCTGTAGGATTCATAGTTA

[0128] TCAATTATTTCTGTTAGAACTGGTTAGAACTTGTAAGAAAAACTTTACAGTTGAAACTAG

[0129] TCCCTGTTGTCCAGGAATTCTTCATGCGCCTTGCGCGTGAACTCGCAGCCGGAACATTTA

[0130] TTTTCCGCATCAATGCGTTTTTTGCGTTCCTCGTCATAGAGCCGCTCATAACGTTCCGCC

[0131] CGTTTCATTTCCCGCCACAGAAAAATTCCCATGACCGCGGCCACGCTGGCCCCGTTTTGA

[0132] ATGTACTCCAAAAAGGGGTTGCCTGACGTGACGGACGCAATCACGGACAGGGCATTAGCC

[0133] CCCAGCAGGCCCGCGTTGACAACAGGTCCGGTCATGGCTTCACTTTTTCAGAGGTTGAAC

[0134] AACGGGCGGAACGTCCGTTTCCGGCTGGGCCTGGGAATAGGAGATATGTCCCTGCTCAAT

[0135] GACGAGGCAGGAGCCGTCTTTGCATACCTCGGCGCGGCCCGGCGTTACGTCCACGGAATG

[0136] ACCGCAGCCGGAGAGGGACATTCCCAAGCCGCCAAGGATGGCGCCAGCGATAACCGCTCC

[0137] GGCTGCATATAGGGCCTTTTTCCACCAAGTGGACGCGCCGGAAGCCTTGAGGCCGAGATA

[0138] GGCCCGAACATCTTCCAGCGCATGCTTACCGATGATCGGGAGGGCAGTATTTGCTACGGC

[0139] AATCCATCCTTGTTGATCGTTTTCCGTCAGGTCTGCCCAGTGAGGGATTGGAGTGTTGGA

[0140] CTCATTGTGTGCCTGGGCTGCATAGTACATGTGCATTTCTCTGGCGATAGCCTCGGCATG

[0141] ATTGCATTGATTATTAGTAGTCATATGATTATGTTGTTATTGGTAGAGGTAGTGAAGTGC

[0142] TTGAAAAAACTCCACGGCGGCGGGGTCGGTCAGGATAAAAGCCGGGTAGTCCGAGACTGT

[0143] AAAAATCCTGCGGCCTTTGGTCTCCGCATGGACGGCCTCAACGGTCAAAGATACCGCATC

[0144] AATCATTGTATAGGCACCATCCTCCGCAAGGGTCAGGACATCTTTTCCCAGCCGTGCCCA

[0145] CACCTGCACCGCCTGCCAGTCCTCACCCATGCCCACCAGAGAGGCAACGACGGCCTGCAT

[0146] GGCCGGGGTCTGTTCCGCTGGTATCTCGTCCTGCGTATAGCGCGCCGGAGGTCTATAACC

[0147] GCCCTTGTCCTGATAAATGGCCGTCAGGGTGAATTCTCCCCACTCGCCGGGCTTCGGAAA

[0148] CTGAATCTGTATCTCTGCGTTATTCATTACTAGAGAGGTATGTTAATATCTTCAAAATCA

[0149] GCCGTTTCTTCGGCTTCAATAGCATTGACGGCCATTGCTTCCAATGCGTGATAGGTTGGA

[0150] TTGGTCAATCCATTGGAATAAAGGTGCCTGGTGCCTGTGCCCGCGTCGGCTGAAAGGGCA

[0151] TATGTTTTCTCATTGCGCGCGTCGATGATCAGAGTGTTTACGCCTGTTCCTGCCTCGAAA

[0152] GCGATGAAGCCGCGGAGAGAAGCTATTTTGAACAGGGTATTGGTACTGCCGCCTCCCAGC

[0153] TCCATATAAAGAGCCGCCTTTTCCTCCCGCACTGCTGTACTCGGCGGCCCGCTCTGCATG

[0154] TAAATAAGCCTGTTCAGCCCGTTCGGCATCAGCTCATTCGTCCCTACTGAAAGAAATACG

[0155] GTTGTCGTCTTCACCTGCCAGTGTCCAACAGACGATACATAAAAGATTTCCCTTACTCTG

[0156] ATTTCATATCCCTTGCGGACAGTATCGTAAGGCGTATTGATGGTAACATCGATGATTTCC

[0157] CCGTGGTTGACGGCCAGATCGTTCCCCGGAATCATAGAATAAGAATCCATCGTCAACCCG

[0158] GTTCTGTTGGTTTTTGAGCCGCGCCCTATGCCAAAAGTAAATTTGGCGAAGGCTGTCGCA

[0159] TTGACCGCAAGGGAAAATCCGCAGACAGAACTGTAATTAAATTGGCCGTTAGGCCCTGTC

[0160] AGGGGAATAACCGCTGACCCGTACGCATTGGACCCGGCAGAAGCCGCGCCCACCGAAAAA

[0161] CGCTGCGTCAACCCGGCAAGAGTACCCTTGGAAGATTTAGAAATGGAACCCGCGACTGTG

[0162] ATCGAACTGGAATTAAGGTATATTGGCTGAACCAACGCTGATACAGCTCCTGCCAATCCT

[0163] GACGTATAAAAGCGATTAACCGCGCCCGTATCCGTCGGCGCTCCCACGGCCAGCGGGATG

[0164] TTGATGCCTCCATTGGCGTTGACGGCCCCGTCAAACGTGCCTCCCGCCGCGGTGATATTG

[0165] CCGTCCAGCGTCATGTTGCCGGATGCGTCCACCTGCGGAATGGCCTCAAGGGCCTGTTGG

[0166] GCCGCCGTGGCGGATGTCGCGGCACTTTCTGCTGATTTTTCCGCTTCTTTTTTGGCATTC

[0167] TGAGCATCGTTCCTGGCATTTGAGGCTTCTTGAGCAGCGGATTCAGCCCGTTCAGCAGCA

[0168] TCCACGGAAGGATCGATAATTTTCACCTCTCCTTCCGCGGCGTCAGGGATCATGATTTTC

[0169] ACGTCTCCAATTGTAACCACATCTTCACTGCCCGGCGGCGTAACACGGGAAGCGGCCTGG

[0170] ATTTCCCCTTTAATCAAGGGGCGCTCCGCCCCATCAGGAGAGGTCATGAAAAGGTCATAA

[0171] GCATGCGTTCCGGCCTTGAGAGGGCTCCACGACAAAAGAGCCGTATTGGCGTCTACAGGC

[0172] TCGCATTTGATGGCGGCGGCGCCAGGGCCGCGGACAGCACAACGGAACTCATGGCCGGAA

[0173] ATATCTTTTGCGGCGCCGGCACCGTCCACAAAACGAAGCGTCACCGATTGGGCAATGCCG

[0174] CTGGTAGTGGCCATGTCAAACGGGGCTGCCTGACTCAGAATCATCATATACTAGTGCGCG

[0175] CCCGTTGCGCTTAAATCATCCGGTGAATCATGGCAAAAATCACCGCCATGCAGGAACAGT

[0176] CCCACATGTGGGAATTATTACCGGACGCAAGCCATTTGAGAGAAACCCTCCCCGTGCGCT

[0177] TGTCTATGGACTCCTGTTTGCTTTCAGCCTGCATCTGCTTGATGTACTCGGCAGAAACAT

[0178] CATGGGGAACCTTGAAATGCCCCGATTTAATCCGCGCGGCAAGAACGTCCTTCGCCCGCT

[0179] GGGAAGAAAAGTTGAAATGAAGGCATTGAATCCCCTCCACCATGTGCCGCTCAAGAGGCG

[0180] CATAAATCAATTTAATCCCCCGCCCCCTGATTTTATGCACGTATTCTTCCTTGACCGTCC

[0181] CATTCATGGAAAACCAGTGATGAAGCCCGCATATCTTCCGCACGGCATCCGTATCAAAAG

[0182] CGCAGTCCAAAGCAACGCAATGATCAGGCACGTTAAACTCCGCCTGTTTACGTTCAATAT

[0183] CCTCCAACGTTTCAAGACGCCCCTCTGAAAGTAAATGGAAAGCCCCTCCGGCATCCACGC

[0184] CGTAAACAGTATGCCAAAAGTGCCCTTTCTGGACATCTACCGTTAAGAAGCGCGCCCGTT

[0185] CGTTTTCCACGGCGTACCCCTCCATGCTTTCCGCCGTGAGCGGGATTTTCAACACGTCGT

[0186] CTTTCTCGCCCAAATCCACAATGTTTTGAGCAAGCCGCTTCTGGATAAACTTTTTCAGCG

[0187] GTTCTATATCCCCGTTCCTCTTCTTTTTATTGGCCAGGATCCATTCCACGGCCAAATCCG

[0188] CCCATGATACCCAGTAACAGGCCAAAGCATTAAAATTGTAGCTTACGATCTCCGGCAAGG

[0189] CGCTCGTATTGCCGGAAAACACGTACTTGCCGCCGCCGGCCAGATAGCGCCGGTTGTGAA

[0190] CAGTATCCTCTATTTCGTGCAAACACCGGGGACAAACCATTTTGACGGATTCCTTAAAAC

[0191] GTTCCCAATCAATATTTTCTTCTTTGCTGTAAATGACATCCCGCCAGTCATAGGCATTCC

[0192] ATCCCTGGCAGCCGGGGCATTGCCAATGGTAATGGTGGATGCGGCCCTTTCCGTACTCGT

[0193] CAACCCAGTCCGTCCCCTTGTCCCCTCCCTGGGAAACCATCAGGATTTTACGGTTGAACC

[0194] GATCATGGTGGCGGGCCAGGAACTCCCGGACCATTCCCTTTTCCCACGCCCACATTTCAT

[0195] CACCAATCAAATACCGGCAGGACTTCGACTGCAACTGCCCCTTGGTAGCGGAGACCATCC

[0196] ACAAGTTCATGTGCGGAAAAAGAATCTCCGTTTTCCTGGCCGCATGCCTTTTCTTCGGCA

[0197] GCAAGTTTTTTAAAGCCTTATTATCCCGGAATGTGGGAAAAAGCCTCGTTTCCGCGAAAT

[0198] CAGAGGTTTCCTGTTCATTCTGGAAAGCGACCAAGGTATTCCCCGCATCATTGGCAATGA

[0199] GGTAATTAAGCAAGCCGACGAACAATGTCGACTTTCCGGAACCGGTGGGAGCCATCAGGT

[0200] TAATGTGCCTGACCGTGCCGTCATTCAGGAATTTGTCAATGGGTTCCAGCAGAAACGGGC

[0201] TCAACGTAAAATCAATATGGGTAGACCGCGCCGATTGCTGGTCTACCACGTTTTCACGTA

[0202] TCCATTCTATCGGCTCCACGTCCATTCCAGCGGATACGTTATCCGCGAAAAGATTCAGAA

[0203] ATCCGGCAGCCTTTTCAACGTCTCTCATTGTTCCGCCCGGCTGATTATCTTGATCTGGGC

[0204] CCTGGATTCTTCCGCAATGGAATACAGAGCGGCCTTGATTTCCTCCCGCAGCATGGGAAC

[0205] CATTTCCGCAGCCGTCAATCCTTCCATCTTCCCCGGCAGTTTATTTTCAAATGCCTGGAG

[0206] AACGCCTTTGGTAATACTGGCCAGATGTACGACAAGCTCATTCACATCTTCCATGTCCAC

[0207] CAGCCGGGCCTTATCCTTTTCAAGCTTCACGCGGGCCTGTTCAGCCTGCAAATTCTTGTA

[0208] GGTAATTTCCGCAGCAAGTTTCCTTTTTCTCAGTTCCGCTACCCCTTCCTCCGGAGCGTT

[0209] TTTCCCAATTTCTGGAGAAGCAGGTTCAGCAATTGGATTTTCCTTCCTCCTGGCTTCAAT

[0210] CAGCTCCCGGATTTGCGCATGCGTCAGACCACGCTTCACGTGGTACGTGATCGTGCTTTT

[0211] ATTGACGCCCAATTGCCGGGCAAGTTCCGCCTGTGAAATCCCTGCCATTATTCCCTTATT

[0212] TACATATTATTATATCATGATATATTGATATGTAAATACACCAAGCATGGAACCGCAACA

[0213] GTCATACTACACAGCACATACGGATTTTGAGTATCCGGAAGAAGGGCAGGAGCAGGAAAA

[0214] CGAATTTCGGGAACGCGCCCTTGAAATGATCCGGATCATGTCCCTGGCCCTGTATCACGT

[0215] CATTAACAGCCGCACTCCCAATGTCACCGCATTCGGCGTGGCATACGCTCTGGGGTTGAC

[0216] GTCTGTGCTCGGAAATGAACGCATGGCCCAGAGGGCGCGGAAACTGGGCGTGCATAAAGC

[0217] GGCCATTTCCCGCGCTGCTTCCAAATTCCTTGCTGAGAGCGGACTCCCGCCCTCCCTGAT

[0218] GATGCAGCAGACGGAACATGCGGCCATGAAGCGCCGCCCTGTCAAAAAGCTGGAGGCACC

[0219] GCGGAAAAAGACGCCGGAACAAATCATGAAAGCATACGGCACGGGAAAGAAACGTTTTGA

[0220] GGAAAACCAGATGATTCTGGATCTGTTCAAAGCCTGAATCCTTCCGGTTCACCCGGGTGA

[0221] AGGATTTCCCACATGATACGGGCGCAAGCTTCCCGGATGGTACAGCCTGATTGCTGGATG

[0222] ATCGGCTCATAAAAATCCTGCTGTCCGCAGTGAACGGCAGACACGCGGCGGGCCAGTTTC

[0223] GCCAGCTTTGATAACTGGCCAGAATCACATGTTCTGATGCCGATCAGGTACTTGCCTCCC

[0224] TGGAACTCAATGACATCAACCCCCTTGCGGCCGTACAGGTACAGAGCCAGCCTGGAAACG

[0225] GTCACGTCCTCCGGATAATCCTGATTGCGTGTCGGGAATTCACGCGTTACCAGGATATCA

[0226] AGAGCATCACTGATGAATTTCCGCGTTACCCAATAACATACTCCGGACCAGGCGAATGGA

[0227] AAGGCGCACTGATATCCCCCTGCCAGTTTCCTGCGCTCAAACAGTGATTTCCGTATTTCT

[0228] TCAAGGGACATCAGCAGTGTATCAGCGTCTATTTTGATGATGGGCTCCCGGCCCGGGATA

[0229] TCACGCATGCAGGACAACATGCCGCGGATACATTCGAGGCCATTCAAGTTCCTGTTGCGT

[0230] TCAAAATAGGTCTTTTTGTAAATGATGTCCCTTCCCCTTGGGATGTCTTTTCTTGCGAGA

[0231] GGGGCTTTTCCGTCGTCAAACAGGTAAATCGTACAATCAGGATCAACCCGGCGGATTTGA

[0232] CCAACGCACAATTCAAGGCATTTATAATCTTCTCTATAACAGAATATTGCGTAATTCATT

[0233] TTTATTAATTTATTGGTGAATAAATTTTGGAAGTCACAATCCATTCCCCCTGCTGGATAT

[0234] AGATTTTCCCGTTTTCGTCCCGGTGCAGGCGTAAGGCGTGGTCAGTGTCGCAGGCCAACG

[0235] CTGTCCAGGAATCGCTGATCAGGGCTCCGTCCCCGCCGCCGGTCGTGTTGGACGTATCGA

[0236] CCTCAAGATCAAGCTGTCCATTCCGGACATACAAGGCAATAGTCGTGCCGTCCGGCAAGG

[0237] TGTGTGGCTCCTTGACATAATCCAGCACAACATACTTCCCATTTTTCCGCAAAGGGGCGC

[0238] GAAAACTCAATTCTTCCGCGTCGCCTCCAGGAGTGACCGCAAGAGACAATACTCCCCTGG

[0239] TGGCCCTGGACAGCTTGATTTTCCACCCTCCCCATAAATCATGCTCCGTATCATCAATTG

[0240] TCAAATCCACCTCGCCTTTCAATCCCTCCTGCGACACACTCAGGTCAACGGGGATTCTTC

[0241] CCTCTTGCCCCGTTGAAGAAGAAACCGTGGAATCAATCCTGACCTGCAGGATTTCCGTCT

[0242] GGATGTTATGGCCATCCGCCCCTTTTTCCGTTTTCCATTCAAGGCCGTCTCCCGGCTGCA

[0243] CCTGCTGAAAACTCCGTGAATCTTTGATTCTCACGCCAAGCCTGACCGGCCCATCTTCTG

[0244] CCCCGCCCTCCCCTTTCCCCTTTTTCTGCACCAGCCATACCTTCAAATTGCTGTCGGCAC

[0245] CCAAATCTTCTTCTTCTTCACTGGAAGATTCAATATTCAGAGTCAGTTCCTTTTCGGTCT

[0246] CTTCTTCCTTTTCGTTCTTTTCTTCCTTGAGTTCCAGCGCGCACAAAGGATCCGTCTTGA

[0247] GCTTGAATTTCTCAAATTCCCATTTGATTGCATCCCAGTTGATACAGGTAATTCCTTTGT

[0248] GCCTGTAGATAATGGATTGGTCGCGTTCCTCAAACTCGCCAACAATAATGTAGGCCTCAC

[0249] CCTCTTCTGCCCCCGGCGGTTCTTTCCCTTTTTCCAGGATTGCAACATAAGCCTCCCATG

[0250] TGTCCGGATCATAGGTCACATGAACCCACACCTTATCGCCTGATTGGAAGGTTGCTTCAT

[0251] ATTCGCTGATTTCCATGCATTCCGTTTTCAGCGATATTTTTTCTTCCTGTTCCTCTTTGT

[0252] ATGCGGACAACAGCACACCCCCGGAGATCTTGCCGGAAAGGCTACCTTCCTCATCATCCG

[0253] TGATTTCCACCAAATCAAAGGCCAGGGGGCGGGAACCGCGCTGATAAGGGGAAATGAAAA

[0254] CGTTCTGCCCGCCGCCACCGCGCAAGGCTGTTTTGCCATGCGCGGCCGCATTGACGGCAT

[0255] TTGTAAGAGCATTGGCCCAATCCGCCTTAATTGGGTCACCCTTGGTGATATTATCCATTT

[0256] TTGTACAGGTCCGCAGACCAGCCTCCGGGGTTGGATTGGGTGTATTCTTCCGTAATTTCG

[0257] TATTCCCGATTGCCTACTTTGTGGACGGAATACCGGGTAAAGAGGTAATCAAGCTTATAC

[0258] TCTTGCACGTTCGTTGCGCCTCCAAACTCGCCGCCCTGGACAATCTTACCCACGCGTTCC

[0259] ACCTTGGACGGTTCCACACGGCGCACCGTGTAGGTTTTCGTGATGACCGTATTGTTCTGT

[0260] GATATTTTTTTATACCCGAGCGTGATATACGGCGTGATGGGCGTAACGGGCAATTGAATA

[0261] TGTCCCGGCGTTTCGTCTTTTTCGCATTTCAGCCGGTACTGTCCATTGCTGCACATGACA

[0262] AGGCCGCCGGACAAATAAACGGCCAGAATCAGCATATCCCCCTCCGGATATTTGCTGCTT

[0263] CCGTCCGGATTCCGCAGCTTGTAATAGCTCAGCAAGGGTTCAAGAGTCATATTTGAACGG

[0264] TCGCTGATCTTGACTTTGTATTTCTCGCTCGCCTCACCTCCGCCTTCCGGATCATCCGGG

[0265] GATGGGTTGCCCTCCTCATCCTCTTTGACACCTTCGGATGGATAGGAGTACGTTACGGTG

[0266] ACCTTGTAAGCTGTCGGAGAATGCAGGGAGATTTTATAACCCGTCGCCACCATGCCGGGA

[0267] GCATTGGGATTGGTTGACGCGGCCGCCCATTTGGGACGCCGGAACCCGGCAATGGGAACA

[0268] ATTTCCGATTGCGAGACGGTACGGTTCACTCTGTATGTCCCGTAGGCCTGGTGACTACCG

[0269] GCATCCGGCGTAATGTCCGTGTTAATCCGTAAATCGTTTGTCCAGATCATGAGCCAATAA

[0270] TCAAAATTCCTTCTTCTAAAATCTGGGAGAAGGATTGCGTGGTTTCCATGTTCCGCCCAT

[0271] AGGTATTCTTGTCTTCCCCCATCTTTACGAAATTGCCGCCTCCCGCAGAAATCAATGATT

[0272] GAAGGGCGGCATCAGGCCCCTTTCCTTCCGTGGTCGTGATGGAATAGGAGCAGGATCTTG

[0273] TGTAATAGTCAGTGATTCCCTTATTAATGATCTTGTTGAACAAGTCAGAAAGTTCCTGGG

[0274] AGTATTTAACACCCCCCAGGCCGAAATCATACGGATCTTTCCTCTTCAGCCTGGAATCAA

[0275] TCTTCAATGATTCAATATCCACGCGGCCCGACATAATTGCTTCATAAGCTTTCTTCAGAT

[0276] CCGGATTATCCTGAAACATGGATTTGAAATCCGGATGGTCAAAGATGCTCTTGCTCGCCA

[0277] CGCCGGCCGTAAATTCCCAGGTGGTTTTCGTGTCTTTGGGGTCAAAGTCCCATGTGTCCG

[0278] GTTCTCCGGGTTCCGGATCGTCAGGCTCTTCATCCTCCTGCGGAGCGTCTTTTTTAAAGG

[0279] TACAGGACGTTTTCCACAAATCCGCCATCCGTGAATGCTGGTAACCGGACAGGGTCAAAT

[0280] CCGTGAAGCCCGGAACATGGCAATGCAGAGAGTAAGAAGGAGTGAATTTCTCCCCGGGGG

[0281] GCGCCACGTAAGAAACGCTTGCCGACCAGGAGCCATCATCAGACATGGATGCTTCAAAGT

[0282] CTCCCTGCCATCCGTACCCGCCTGTGTGTTTAATATCCTTGTTCATGTCAGCTAAAGAGG

[0283] GCAACACCGCCTTTCTCGGCCTTTTCCGCAATCGTCCTGAGGTATCCGTTGGAAGTCTGC

[0284] ACTTCCTTTGTGAGGGAGTCCACGGCAGCGGAAAGGCCGGCATCCCCCTTTCCCACCAGG

[0285] CTAAAGAGGGAACCGAAACCAGACCCCACACGGGATACGCTTGAAAGCGTCTGCTGTACT

[0286] GCGTCAAACCGCGTTTTAAACGCATCGGATATTTCCTGCGCCCGCGCCCTGGAATCCTCT

[0287] TTTTCCTTTTTCTCCCTTTCCCTTTTGTCCTCAATCTTGTATGTCAGGGAAACCTTCTGT

[0288] TCCGCCAGCCCCAGGGCGTCATGCTCATTCATGCCGCTTTTCACGAGGGATTGCATTTCC

[0289] TTCCGCACATCCCGCTGCTTTTCCAGTTCGCGCCGCTTTTCCTTTTCGCCGTCAAGTTCG

[0290] GCCTGCATGATGGCAATTTCTTCCTGCGCATTCTGTCTCACATCATTAACGGCTCTGTTC

[0291] TTTTTATTTAATTGAGCGTTAATTTCCCGGATGGAGTCAGCCACGGATAACAGCTTTTCA

[0292] TATCTCGTGGCTGCATTAAAATCTAAAGCTCCTGCTTGGAATTCTTTTTCAAGTAATTTC

[0293] AATTCCTCTTCCAAATCATCCAAGGAAATATGTTGAACAGCGGCTAATTCGTCTTCAAGC

[0294] TGTTTCTTTAGACCATCGACTGATAAATCATCCTGTTTCTTTTTAGGCTTTTGCTTATTT

[0295] GCTTTATCTCTTGTTTCATTAAGCTTCTCCATTGCATCGTTCAAATCAGCGGCAGCCCTC

[0296] GCTTCCTTCTGATTGTTTTCTACGATCTCTTTAATTGATTGCTTCTCATTCTCCCTCCAC

[0297] AAAACTCCTTCTTCTATTTCCTCAAGAAGTTCTTGACGGGCGGCAATCATCCTGTCAATA

[0298] GCCTCAATTTCCGCCTTTGCTTCTGCACTGTTTACGCCATCGTTTGCAATATAATTCTGA

[0299] CGGATGCCGCGTTGTTTAATAAGATCATCCCATTCTTTACCTAGCCGCTCTTGAACAAGC

[0300] TCAATTTCATCATTGCTTTTGGCCTTGGATATTTCTTCCCTGATTTTATCAGATGAACCG

[0301] TAATCAATCAGATTTTTCCTTTGGTCATACGTCCCCTCTTGATATGGATTTTCCCACTTT

[0302] CCATTCACAAAAATCCCTGTAAGACCAGCAAAACCAGCCTGAACTCTAGATAATATGATC

[0303] TCAAAAAGAGATAAATTCTCTTGATTCATGGCGTAAAAATGAAAACCAACCTCGGAGGCC

[0304] TCTTCTATTCGAGATGTCAGGTACTGAAAAAGCCCGGCCGTTTTTGAGACATTATCACTC

[0305] ACCAGATCCCCCATAGTATTTTCAAAAGTAGCCTTGAGCGTTTTCCATGAACCGATGGTC

[0306] GTATTAGACAATGCGGAATTCTTCCCGGAAAATTCCGCGTCACGCATTTTCTTCATGGCG

[0307] TTCACGTACTGGTCAGCGGAAATTCCCCGCTTCTGGGCGAGCTCAACAACCTGTCTTTCC

[0308] GCCACGCCCATTTCTTCCGCCAAAGCCTTGATAATGGGAATGCCCTGATTCTGCAGCTGG

[0309] TTAATGGCCCGTGAATCGGCAAATCCGTTTGCCATCACCTTCGCCCATGAATTGGAAAGC

[0310] TGCTCTGCGGAAATCTTTCCGGACGCGGCAATATCTGCAAAAACTTTCGACCAGCTCATG

[0311] ATAGCCGTATTATTTGTCCCAAAATGCATGGCCAGGTTTTGAGCGGCTTTCCCTAAATCG

[0312] TCCAGTCTAACAACGCCATTTGCCGCCAGATGATCAAGCTGGGACTGCAAATCCCTGGCC

[0313] GACTCAATATTTTTTGTATAGGAAGATAACTGTACCGCCATGTATTCATCATCCATCGCG

[0314] TCATTGATTCCTGTTTTGACAGCGCTCCATGCCGCCCCAAATCCTGCCGTGGCAGCGCCG

[0315] GCAGCCGTAATTGCAGGCAATAACTTCTTGGAAGACGCCCCCAATCCCTCTACGTCAGCC

[0316] TTACCGCCCTTAGCAACACGCATATTTGCCGCCTGCCCCGAGGTCTCGCGGATCTTTTCC

[0317] TTAATGTTCTCCAGTTCTTTGAGCAATTCCGAAGAATCTCCGGAGAATTTAAATTTTACG

[0318] GCACTCATTCTTCAACATCAACAGGCTTGTTAAACAGAGCGTCCAAACGCTCCAAATCAG

[0319] CATGATCCACCTTCCGGGCATGGGTGTAATAACAGCTTACGCCATTTGCAACACACTCGC

[0320] TCATCAGGAACTGCAGGATCATGCTGTAGGGAATATCATATTTAATGTAATTTAAGGGAT

[0321] AGCCCGTTTTCGACGCAATCGCCCAAACCACCTGGATAAAATAATCCGGCCTGTCCCCTC

[0322] CCGTTTTTTTTTGGAGTCCTCAACAGTAAAAGCGGCTTCATCCACCTGCTGCGCCTGCAG

[0323] GCCTACCAGGGAGGAAATACGCTGCATTTCATCAGGGCTTAATTCATATTTGAATCTTCG

[0324] GACGGCTTCCTTAAACGCTTCAAGGCTCATGGACTCAAGCGTTTCAATATCCTCCGTATG

[0325] AATATAAATATATTCCACAATGCAGCCCAGATTCATGCTCCCGTCCATGAATCCACAGTT

[0326] AAACCGCTGCATCAGCTCCATGCTCCCAATAGACAGGGGACGGAGCTTCAAGGCTTTTTC

[0327] CTCATCTTTCATCATATTCCAACACTCAAGAGTTCAAACGCCCCATGATTTCCATTTTAT

[0328] CCTCTTCCGGACAATTCTCCGGAATTAAGGCGACCTTCTCCCCATCCCTGGAACGGATGC

[0329] AGGCTATTTTCCGGAATCGGGAAAGGCCAGACAGAATGCCGCGGCGGCAATCAAAGGCCG

[0330] CCTTGACCAGGGCAAACGGATCTTCCGGATTGTTCTTCACAAAATCAGGATCCTTCCACT

[0331] TCCTGATCATTTCCCGCGTCTCCAGGGTATCCGTAAAATAAAAAACCTTCCGTTCTCCCC

[0332] GGTCTGTAATGATTATCTCCGCCCGCTTGGCGATAGGGGCTGTAGCAGCGAGCAAAGCGC

[0333] TGGCGAGATTAATATCTTCAATATTGATTATATTATTCATGTTCTGTCTATTATATTATT

[0334] TTTTGTTTGAGGGGAGGGGAGGCGTGAAGCTCCCCCCCCTTAAATTTGTCATTCGGAAGC

[0335] AATATCCGGATTCAGAGCAACGATATTAGGACCATAAGTAGCTTTAACCGTGAAACTCCT

[0336] ATAATTCTGCGTCTCCATCTTCACATCCACGCTCTTGCAAATGGCCTTAGTCACCTTTGC

[0337] CGGAATTTCTTCACCGGAATTCTTGATGCAGAGATTCAAGACATCAGCATGCACATCAAC

[0338] AATATCGCCAATGTGGGGCACTTCCGTGCCAGTCTTTCTCATATAGGCTTCCCAGGTAAT

[0339] TTCACAGCGGGAATCATAAAGCGTGTTCCCCTCCATCAATCCCTTGTCCCCAGTAAGTTC

[0340] TACATCACCTTTGGCCGACCATGAATAACTAACGGAAGAAATGATTGCTCCGAATTCCTG

[0341] TTCACCGGAATAATTGAGCCCGAAAATACCTTCTCCGTGCTGGATAAAATCAGATGCTTT

[0342] TGCCATACAAATGGCCGTGCGTTGCGCTTCCGCCCCCCCCCCCCAAAAAACAGGAGAATG

[0343] AACCTCCTCCACAAAAAAGCCCCGCTCATATTTCATGAACGGGGATGATTCAAATAAAAT

[0344] CGAAGCGTAGAGAGAAAACCGGATTACTCGCCGGATTCCGGCTGGAAGCGGGCCGCCTCC

[0345] GATTCTTCAGTAATTTTGGCCGGTTCGATACGGAGGAGCTTTTTCATGGCGCGGCCTGAG

[0346] TTACTCTCCGTCATCGGGAAAAAACTGACGTCATTTCCCAAATTGTAGGAAGCCAGTCCA

[0347] TAAATGCTGTCAGCCGGAATAAAATCTTCCGGCGTACCCAGCAGTTTTTCTACCTCCTTT

[0348] GAAGTCGCAAGGACATAACTCAGATTACAACTCGAATACAGTTCTGTTTTAGGAGAGCCG

[0349] CCCACAGGGAGATTATTCGCCGCCCCTCTCCACTGAACCAGGGCTTCTACAGGAATGTCT

[0350] CCCCGGAGGGACTGAATGACGCGGGCGTAGAATATCGTCTCCTGGGTAGTTTCCACCGTC

[0351] TTATAAATGACAACCGCTAAGACAATATCGCTTTTCTGCAGGAGTCTGGCTATACCACGG

[0352] AGAGAAATATCCCCGGACACTTCAGCCTGCGTGGCAACAGGAGACGTAGCACTAACCGAT

[0353] AACGCCATCAGACCGGAAAGCAAACAGCAAAAGAGAGATGTTTTATCCATAAAAGTATAT

[0354] ATTATTTTTTCCCCATTATGTTCAAAAAACGGGGCGAACGCTCTAACAGCATTCGCCCCG

[0355] TGAAAGAGAAGTTTATGAATTATTCAGCGTCTTTCAATGTGGACCGATTGATTAACCAAA

[0356] TAGGGCCAGTCAGAAAACTGCCAATGAATATGTTGTCTCCCGGACACATTCCTACATGGC

[0357] CATTTCCTGCAAACACAATGATGTCGCCCGGCTGCGGGCTGCAATAACCTTTATCCTGGG

[0358] CGTCGGGATAATCAGGGTCGCCCAAGAGTTTTTGCACATAAGCAGCCATATCCCGTGCAC

[0359] GGGTGATGACATGGGTTTTACCTCCTATTGTATCAGCGTTCGCCTTGTCTCCTATGAGCT

[0360] CCGCTCCTGCTTCACCCTTCAAATCCTTTCCGAAGGAACTTAGAGCAATACTCATTCGCA

[0361] AGGCACAACTGTTATAATAATTGCGCGATTGTTCCGGAACATTGTCCTTAATTTCCTTTA

[0362] AGTGAGCCTGGTAGAGCCACCCCCCAATATAAGCCCAAACCTGTCCAGGATCAGGCATAG

[0363] TGACATAATCCACCACATTGTAACATTTCAGCAGTTTTTGGGCATCCTCCATCTTCCAGA

[0364] GATTCGTGGGAGCATCAATTTCCCAAAGTTCCAGGTCAACACCATCCATTTGAGGAACGA

[0365] ATTCTTCCGCATTGGGATAGGGAGCAATTGCATCCGGAACCTTCAAATTTTCATGATGGA

[0366] GCTTGACATAATGGTAACCTTGCTTGAGCTCCTTCTTACCCTCCGCTTTCTTGGATGAGT

[0367] TGCTACCGTTGAGTTCCACCACTTTCTGGTTGTCGATTTCCAAGTAACCGTTGTCGTCCA

[0368] TCTGAATGGTAAAGGCACATTCCTTATCGAAGGGAATACGAATGTATCCTTCCCAATCAT

[0369] GCGCCCCGGTTTCTTCCGTCTCATGCCAGACGATCTTGGGTGCGTCCTTTTTCTTCACTT

[0370] TATAACGAGGATTATTCTCGGCGTCGAGCGGGTGGAATGGTATTGTTGATATAGCCATAA

[0371] TGTTATGCTTTCTAATCGTTTATTGTTGAATGAACTGAATATTCAGTTCATGAAAACTTT

[0372] GAAAAATTTCTCGCTCCCCATCAACACGTTTAACGCGTAGTTCAACCCACATTCAGAACG

[0373] TGAGTAAAAAGACTAAAACGGCACGATAACCAAGCCGGAAATAAAGGATATGAAGGGGTA

[0374] CAAGAAGTACAACCATTGATTATCAAAATAGTATAAAACTATCTTTCAATATAACGCAAT

[0375] GTCCAGGTAGTTACCCAGTCATCATCATCCAGCGTAGCCGAATAATCCGTAATATTGAAA

[0376] AAATCACACTGTTCACCACAATCCAGCCGGAATAACGTCTCCACAAGGGTTCTTTCCATT

[0377] TCCGAATCAGACCGGTTATTGATCAGAATCATGGAAAGCGTATATTCCCTGGCGGGCAAA

[0378] GGCGTCCAAGGCTGAGTATACCCGACCATGCCTATGACGACGCAGGGCTTCCGTTCTATG

[0379] TCGCTGTTATTCCCGGCTTTGTAGACAGTAAACCCGGAATTCCTCAAATGACTGATGACA

[0380] CGATCCATAATCATACCCTGTTTCTCCTTTGCCACTCATGGTGAAATTTCAAGTTCGTGG

[0381] CCTGCAATGCCTTGGTTTGGACAAACGGTACAAGCGCCTTAAGCATAGCCTCCGAGGCAT

[0382] AAGACAGATTATTTTCTATTTCGACACTTCCATCAGGCAACTTCTTGATTGTCCCGTTTT

[0383] TCCCATGCCGCGCAATCCATTGAGGCGCACGAGCAGCCCATCCATAAGAAGTATTAATCA

[0384] GCCACCCTGATTTTGCCTTCCCGATATGCTTGAATACCTTATTCTTATAGTTCTCCGTAC

[0385] TCTTTCTATTGAGCACGACATTCCCAGTAAAGGAGGACACACGACCATTCTTCCGGGCAG

[0386] AAGTATGAAATTCCGGGCTGTAACTCTGATAAAAGCGCACAACCTGCCCAAAAACATCCG

[0387] TCGCCACTTCGGCAGCCTTTTGAGGATCTGCCTTTAAAGAAAGCATGCCGGCAACCTGTC

[0388] TTGCCCTTTTTATGTCTCTTTCCTTTAGTTGGTTGATAAACGATCCAACACCGCGCCCGG

[0389] CGCGGGAAACGTCCCGGCTGACAGACCCTTCCCCGGATTTTCTTCCGGAAACCTTTGATT

[0390] TGCGATTTTTTCCCAGCGGAAGAGTGGCGTTAAAGCACATCCCGCTAGCGGTATTGTAGG

[0391] CAAGCAACGTAGTATATAAATTTCCGAAACGCTCAATATTCGTTTTCACCTTTTTCAAGG

[0392] CGCTATCCATTCCTTTCACATCTATCCGGACTTCCATGCTACTCATATAAATCGGCTGAT

[0393] AAAGTTAATACCTTGTAAACACAATTACTTGTGACGGCGGTAATCTTGAATTTCATCCCT

[0394] TCGCACCCCACACGCCCCCCAATCTGAAAAGCGTCCCCTGTCTGGATGTGAATGATGCGG

[0395] TCACGCTGGTTGCAATACCCTCCTTCCTGGCGTTCAATGGTTATGTCCCGGGAATTCATG

[0396] TACCCCCGGTACGTCTGGCCATTGTAGACAATTTTTGTCTGTGGTAATACCCGGTTAATA

[0397] CTCCGGTGAGCAACGTTGGCAGCCTCTGTGAATCTCATACAAATGGCGGTGCGTTGCACA

[0398] AAAAACCGCCCCGGACAAATCCGGGACGGTAAACACCAGCATGAATACAAAGAAAATCAG

[0399] CCTTCATCTGACGCAACAGCAGCGGCAACGTATCCTTTCACCTTGTCAGCCTCGAGAATC

[0400] TTTGCCCCGAACATCAGGCCCAGAGAAGCCCATTCCGCGCGAGTTTTCAAATCAAGCCAA

[0401] GTGCAAAACAGGATAGAAAGACCAGCAAGACCTTCCACCTGAATGACCTCGGAATGGATA

[0402] TCATCCTTGAAAGCCTCGTCGATCTCCGGAAGGGCGGAACCAACAGCAACCGCACCTCCA

[0403] TCATAGGAAATACCGACTACGTTATCAGCAAAGACGGAGGTCTTGACCTTATGGATGGAA

[0404] TCAAATCCGTAAGCGCCTCCATCAAGCCGGAATTCCGTCGTATTTTCCGGAAGAATAGCT

[0405] CGGTAATAATCAGGCGCCAGCAAAAGGCGAGGTTCGTCGGATTCTACGTCTCCGCTGAGG

[0406] TTGATGCACTTACGCTTTGTGAAAGCGTCAGCCGTACCGATAACCTTGGCTCCATCCGCT

[0407] GTAACAAGGGAAAGTATGCGCTTGTTCAAGTTATTGGCAAACGCCGTAGCGTTCGTTTTT

[0408] GCCAAATCCGCCAATTTCCACCCATTCCGCTTTTCGGCGGGCGTAATGCGCCAGGGTTGA

[0409] TGGAGTTCAGTAAGTGACAGGGGGATTAAATCCCCTTCATTTTCTTTATCCGTCTGGTAA

[0410] TCAGAAGGGTTTTCAGTAATATTTCCGGCGGCTTTTACATTCTTGAATTTAATCTCAGAA

[0411] GCATACTCCATTCGGAAATCCTTCTTAAAGTATTCCAGCGGAGCCAGAGCTTTACGCAAA

[0412] GAAGAAATGACTTCACTGTGAATAACTATCTTGTGAGCATCTGAAAGAGCCATAATTCTA

[0413] AAAAATGGTTTTAATCGTTAATATTGTTATTGATTACTTGCAAAGTTTTGCCAACTCATC

[0414] CTTATGAGCAGCATAAAACTTCCTCTGTTCCTCTCCGGAAAGAGCTGAGTACTGCTCCAG

[0415] GATAGAGGAGTTTTCTTCGCCGTCTTCCGGCTTATCCACTACGGCTTTATGTCCCATACC

[0416] AGCCACCATTTCAGCGGCCTTCTGGCTCGCCTTCGTATCGGCTTCTTCCTCAAGCTGGTT

[0417] TTTCTTGAGGGCAACGGATGCGTGGAGCTTGACATTCTCCTTTTCCAGGGCTTCGACACG

[0418] CTCACAAAGATCTTTAATAGAGGATTGGATTTCTTTAATCCCATTCTTCATTTCTGCCCC

[0419] AAAAGGGGCAAGCTCTTGTTTGAGCGCATCAACGATATTTTGATCAGACTTGAATAGTGC

[0420] CATACTTATGACAAAACCGTTGCGTTTTCATACCACTCCAGGAAAATCGACCCAGGGTAT

[0421] TACATCATCAATCAGGCCCAGGGAAACAGCATCTTCCGCCACAAAAACCTTGCCGGAAAA

[0422] CGCATCCGCAGATAAACCGGGACGTCCTGAAGCCACAAATTCCTGAAACCGCCCGGCGAT

[0423] CTTGTTGCACAAATCCCTGTAATAGGCGATTGCCTCCGTATTCATCGGGGTTTCCGGATG

[0424] CTTGAAAACAGCATCATCATTCGTGATGGTCAATATTTCCTCACTCTTGGCATTCCACAG

[0425] GCTGATGACCGTCCCGATACTGCCAACCAGCGCCGTTTCCGTCACGACCACTTGATCACA

[0426] AGCGGAAGCCAAATAATAGGCGGCAGAGCAGCACATGCCTTTGACATAAGCCACTGTTTC

[0427] CACTGGCAGAGATTTAATCAATTCTGCTGTTTCATGACAGCCTTGCACGTCACCTCCCGG

[0428] AGAGTCAAAAATGAAAACAACTTCATCAATGTTCCCGGCAGCACCCTTTATTTCCTTCAC

[0429] GATGGACTGATAATTCGTGCCGCCCAGGACTTCATTCAACGGCACGCCGTCACCGTATAG

[0430] AGGCCCAATCACGTCAATCACCAAGGATGACCCAATTACCCGCGCTTTTTGGCGTGTCTG

[0431] GATGAAAAATGAGAAGTCAAAATCCGGATTCCCTGCGGCTTTCAGTTCCAGGCTCTTCCG

[0432] CTGGCGAATCTTCGCCAGCCATGCAGTTTCTTCACAATATAACGGTTTCATCACATTAAA

[0433] ACTTCCTGATATAATCCGGAGAGATTTCTACTCCAAATTCTGCTTCAGCTTCACGGCGCA

[0434] ACTTCTCCGCAATGGCGGCTTCCCTGGCACGTATTCTCAAATGGTCCTCAACCTGCGTTC

[0435] CGTTTTCCTCACAGATTTCTGTGAGGTTTTTAATACCGGCATTGTACTCTTTCAACTGGC

[0436] TGTTGGAGTCGCGGCCCATGTCTACGCTTGGACGCTTGGCGCGGCTGAACATCATGTTCC

[0437] ACCAGCCTTCCGCGTGCGGGATGTCCTCCCGCTGGATACCTACGGCCAAGATATACTGTA

[0438] CAAGCAGTCTGGCAGCATCTTCAAGCAGCGTGGCCCGATCTCGCGTGGTATTGTCAAATT

[0439] TGGACAGAGACATTCTGTTCCCAACGCCGCTTGAGTCCGGATTGACCACGATATCATAGG

[0440] GCACGCCCACCCCGGATAAAACGCCCTTCAGAAGCCGTTCTGAAAAACTCATGTAGTTCG

[0441] GAGAGGGGCGCTCAATCTTCAGGGAGTCAAGGGAATCTTCAGACTTCAAAAATACAATTT

[0442] GCTTATCACCAGTATTCCGGATGAGCACCTCCCCATTATTGGACGGTTTGTCATAAGCCT

[0443] GCGTTATGTCAACTTCTCCCGTTTCGTTCTTTTTTACCAAGGCAATTTGTGACGCAATCT

[0444] TCATGGCCCCAAGTTCGGCTGTATTGATATCATCAATGTATCGGAATTCCTTGAGGCCAT

[0445] GAGAGAAGAGAGGCTCGCCCCGGCAGGTGGAAAGAAAGTCGTCATCCACGATATGAAGTA

[0446] TTGAGGATGCCGGGATGATTTCACCTCCCCCTGGTTCGTCGCCGTCCAGCCAATAAGCGA

[0447] TTTCACGTCCCATGCGATTGGTTATGACGCCGTGCAGCACGTTCAATCCCTTATATTTTC

[0448] CGGAGTCAATAAGGCCATTTTTAGGAGAGCAAACACGGTTTGCCCGGATAAATTGCAGTT

[0449] GAGGAAACCCCGTCTTGCTTTCCGTCAACATTACAAAGCAATCCCCATCAATGTCGATGG

[0450] CGGTGGAGATCAGATACAGGAGCGAGTGAAAATTCTTTCCGTTCACGCATGCAACTTTGT

[0451] AAAACGCCTTGATGTACTCGTCATACTTCGCCGCAACTTCCGGATTCTTGCACAGAGATT

[0452] TGAACAGAAACGCTTTCCCGATAGAGTATTCAGCCTTCATGTTCACGGCGCCTTTTACAA

[0453] CGCCATTATTTTTGTACAACAGACGGGATGCGGAAATAAGGTTCTTCCGGGTAAGGTTCC

[0454] CGGAAAGCTGTTCCGGTTCTTTGAGATAATTCGGAAGCCGCTGCAGCCCGTCATCATAAC

[0455] TGCCGCTCTTGTAAATGGGGCGGCTTGAAAACGGCCGGCCGAACTGATCCAAAATCTGGA

[0456] CATTTCCTGCTTTCATCGAAACACTATCCTTGCAACGTTATTCCCTCCTTCAAACCGGCC

[0457] ATGCTCAAGAGAACGAATAGCAATCCCCAAAGCTTCCACCCATAAATCAATAGTCATGGA

[0458] ACCTGGCGCGAACTGGAATGAACTGTCCCCCACGTTCCCTTGAGTGATTTCCTTTCCCTG

[0459] GTTTTCAATGATATCTTCAACAGCCTTATCCAAAGCTTCTTTCAAACGGGGTATGCGTTC

[0460] ATCAGCGTCCACCGCCCCCCAAGCATAAAGAGCTTTTGCAATCCGTAACATACAAATGCG

[0461] GCCCCGTTGCGTTTCCTCTACGGCAGACACGCAGGCTTTTACTTGAACCTATACTTTTCC

[0462] CGTTGCATTGTTGGAATAGTTGCATGAAATTTTTCAACTCCATGCATGCAAAAACACAGA

[0463] TGACCCATGACCCAGTTCGAGCCTCTACGCTCAAGGAGACTCCCTTTTTCGGCCTATTTC

[0464] AGACTATTTTTCATCCTTCCGTGTGCTCAACATTATCACAATAATGAACACATTTTATAA

[0465] TATATTATTCTAAATAAACATATTAATTCCTACCGGGCCTATCAATTACGCCGGGCCATT

[0466] TCCGCATTGTCCACGGAATTGACCGGAGAATCTATGGAAATGCCCTCTGTCAAATTATGG

[0467] CTTGATGACCACATGCGGCGCATTACGCGCAATGGGCTTAAACCCGCATCCATAGCGAAC

[0468] TACAAACAAGCCTTTGACAAATTTCGCGCCTCAATGGGAGAACGTATCAACCTGCCTCTG

[0469] GATCGCATTACTCCTCTGATGCTGGACGATTTCAAAAACCATCTTCTTTCCCGTGTCTCA

[0470] CCATCTACCGCCAATATTGCTCTTACGCTGGTTTCCGCGGCGTTCCAGGCGGCAGTTGAT

[0471] TATAAAATTATTGAAACCAACCCCTTTACGGCGATTACCAAGCCTCACAAGGGGAAAGCC

[0472] GTCAAACGGCGGAAATTCGAATTGGAAGAGCTTGAAAAGGTAATGGCCGCATGCAATCCG

[0473] GAATGGCGCTCCATGGTGAAAACGTGCCTCTATACGGGCGGTCAAAGATTGGGAGACGTG

[0474] GCAACGCTCCGGTGGTCCCAGGTTGACGAGAAACGAGGCGTTATCCGGATGACCACGCAG

[0475] AAAAAGGGAAAGCCTCTGATGATTCCGATTTTTCCGGCGCTGAAAAAACACCTGCAGCAA

[0476] CGGAAGAAAGAAGCTCCTGGGGACTTCCTGCATCCTGAATGCGCGAATATTTTTGAAAGC

[0477] AAGGGATCCGGACGCCTGTCAAATATCTTTAGCCACATCCTGTACCAGTGTGGCCTTATT

[0478] GCCAAAGACCCTCTGGCTGCAGGCAAAAAATACAAAAAGCAGGAAGGAAACGGCACAGAG

[0479] ACGCGGCGCCACGTCAATGAATTGTCCTTCCACAGCCTCCGCTATACGGCAACAACCATG

[0480] TTACATGACGCCGGTGTTCCCCCTGCTCTTGTGCAAGCCATTGTGGGGCACGATTCCCGG

[0481] GAAGTCCATGAAGGATACATCGACTTTGGAGCCAAGGAGTTTACACAAGCCCTTGAAAAG

[0482] CTTCCCAAATTGTAGCACTACCTTAAATAGTGATATCATGTTTTAAACCTCCCTGTAAGT

[0483] CGTGGCGGCAGCGTCCCATTTCAGGTCAATGTAACCAATCTCCCCGAAGCGGTTCTTGCC

[0484] GACGATGATTTTCGCCTCTGCCGGATCTGCGTTTTTGTCCATGACGTAAGGCCTGTAGAG

[0485] CAACAATATCTGATCAGCGTCCTGTTCGATAGACCCTGAATCCCTCAAATCGGACACACG

[0486] GGGAACCCCGGCTTCCTTCCCGGCGCGTTTCTCCGCTTCCCGGTTCAACTGGGCCAACAC

[0487] GATCACCGGAATATTCAGTTCCTTGGCCAGGGCTTTGAGGCCGGCGGAAATCTCCGACAC

[0488] CTCCCGTTCCCGGGACGCCTGGCGTCCGGTAGGATTGGCAAGCTGCAGGTAATCCACGCC

[0489] GATGCACCTCACGCCGTGGTCCGCCACCATGCGCCGGGCGGTCGCCTGGATCTGGTCAAT

[0490] CCTCAAGGCGCCCCGGTCGTCCACGAAGAAAGGCAGGCTCCTTACCTTGCGAACCGCGTT

[0491] GGTGAAAGCATCCTGCTGCCACTTGGTCAGCTTGATGCCTCGGCGCAGATTGGCCGCATT

[0492] GATTTTGGACATGCCAAAAAGCGTGCGTTCAAGCAACTGCTCCTTGGACATTTCCAGGGA

[0493] GAACATGCCTACTGGCACCCCTTCGGCGGCCAGATTGTACAGAATGTTGGTCATGAAGGA

[0494] AGTCTTCCCCACGGCCGGCCGGGCTCCGATGACCACCATAGCCGTGTTCTGTAAGCCGTC

[0495] CAGCATCCTGTCCAGGGAGGGGTAGCCGGTAGGAAGCCCCTTGGTCTGTCCTGGATTCTT

[0496] GATGCGAAACTCCAACCCTTCCACCACCTTCTGTGTGCCATCAGCCATACGCGCCACTTG

[0497] GGCCACTCCGTAACTTTCCCGTAAGGAGGACATCACCTTTTCGGCTTCCGCCAGCACTTC

[0498] GTCTTTGCTCAGGGTTGGGTTTTGAAGATTTTCCAGCCCGGAAATGAACAGGGATTCCAC

[0499] ATCCCGCTTCTTCTTGGCTTCCACCAGAATCTTCACGGAGGGCTCGAACTGGAAATGGTA

[0500] GGCAAAGCTGGTTGAAAGCTCGACAAGCCCGGCGTGACCTCCTACGGATTCAAGCTCGCC

[0501] GGCGGCTTCCAGGTGCTGGATCAGGTCGGTGATATTGACCTTTTCTGGAGTCTTGGCCAG

[0502] AGTCTCAAAGGCGCTCCAGACCTTCTGATGGGCCAGAAGGACAAAATGAGCCTTCGTGAA

[0503] ACCTTGCTCGATCAGGGCGGCTACCTTGTCGGCGCCGTCAATGCAGTTGCCAAGAACGGT

[0504] CTTTTCGGCGTTGAGTTGTGTTTCTGTAAAATGCATGATGGTTTCTCTTTTTGAGGGTTA

[0505] AATATTTTTTCTAAATTTTGGATCGAAGGGGTCGCTTTCCTCTCCGGGTAGTTGCCCTTT

[0506] GGCATCAAGATTTCTTGCCCAGCGCAGGGCGTAGCCCAGGGCATTGGCTTTCCAATCGGC

[0507] AATGGGGCAGGAGAACTTGTCCACCCAGCCGGAGCCGCCCCGGTCCGCCCAGTATTGCAG

[0508] GGCGCATTCCGAAACGTCCCTGGGGAGCAGCTTCAAGCGGCATCTGGCTACCTCGTTTTT

[0509] CAGGTAGGCTTCCACCTCTTCCGGTGAGGAAGGAAGAGGAAGTGAAGAACGAGAAGGAGC

[0510] CGCAAGCGAGCCGTCAGGCGCAGCTTCCTTCGTCTCCGTCTCCGTCTCCGTCTCCGTTTC

[0511] CGATTTCGTATACGTATTCGTATTCGACTCCGTCTCCGTCTTAGTGTGCATATGCTGCGC

[0512] ATCTGCTAAGCATGTGCTGCGCATATGTGCATCAGGCGCAGGGTATTTGCTTTTCTTGCT

[0513] CCGTACCTGTTGCCGGAAGTTGGTCACTTCCAGATATTCTTTTCCCTCGATGGAGTACAG

[0514] GACGACGAGCCGGGCTGCCTCACAGGATTTGAGGCAACGTTGAACGGAATCCTCCCTCAT

[0515] GGAGTCGAGTTTCAAGGGGTACAGGGCAGAGCGTAGAACAGACGAACGGGCGTCAAAACG

[0516] TCCAAAGTCGTCTACAACGGAGAGCAAGCGGCGGAAAAATACCTCCGCTTCCCAGCTCAG

[0517] AGAATTAACCGCTTCGCTGGTTAAAATCCCTTCTCTGATTATTCTATTTGGCATATCAAA

[0518] AAAAGTGTCAGTTGGGGGTTTAAATTTCATAACCACTTGATGGTGCTAACTGTATTGCGG

[0519] CTTTTAACCCAGTGGAACCATGCGTATGCAATGGCTCCGCCTTCGGTTTTGGCAAAATCC

[0520] CCATTTTTCGCACAAACCCTCCGATCTGAAAACACCCACACATCTGCGGGTGGCGCTACG

[0521] TCGAATAACCGGCGCCTGGCCTTACCCTCTAAAAATTGGAGACGCAACAGCATCCAGATG

[0522] TTGGCACCCGGACTGGCACAGATAAGCGCTCTCTCAACAAATTCAAGGGCCGTTGCATAG

[0523] GGGGGATTGGTCATGATGTCCACATCATCATCCGGCACAGGGGAAAACTCCCAAAGGAAA

[0524] TCTTGCAGGCTTGAATCAGGGCAACCCCTGTCCACTATATCGGTTGCGTACACTTCATGC

[0525] CCGCGTTCCCTCAAAACGTTGACAATGTGGCCGGCCCCGCAGGCTGGTTCCCATACATAT

[0526] TTCCGGAGGGGCGCACCGGCGTTGAGCAGATCCCTCACCATGTCCGGATGCGTGGCGTAG

[0527] TAGTCTTCCCGTTGCCGATTCTCGGTAGTGTGACTGGATGCGCCCAGCGTGGCGCCAAGG

[0528] GTTCGGTTGTTACCTGTCCAGTCCTTCATGCCCCCTCCTTTCCACGCTGTAATCTTCCGA

[0529] GCAGACCCCAGAATGCAGTTCCGCACACCTCTTGCGGACGTAATCCAGCACCGCGCCGGC

[0530] GTCCTGGTGTTTGACGTGGAGCAGAAAACCCGGCTTGCCGAACTCTTCTACCCAAAACTC

[0531] TACTTCCGGGCCTGTAAGGTCAAAATACACGCCGTAGCGGCCTGTACGCCCCAGTTCCCC

[0532] CTTGATGCGGCAAAGCTCCTGAATGAAACGGCGCTCTTCCTGTTCTGGTGTTTCCATAGG

[0533] AAAACAAGCTGTCTCATTTTCAATTCCAGCACGGATGCCGCGGCCTGGTTGGTCAATGTA

[0534] GGTGGATTTCACTTTTCTCCTCCTTTCTTCTCTTATTCAAACATAACCACCTGGAGCTAC

[0535] CATGTGCCTTACCCGTCCGGCCCCGTCCACGGAACCGCTCTTAGAGATCTTCCCAGGAGC

[0536] TTCTTCAAACTCGAACACGGCCTGCTTGCTCAACTCCACCATGGACCATACATCATGGGG

[0537] GGAAACCAACAAAGCTTCCCCGCTAAAAAGATGCACCAGTAACCTGCTGGGAGTAGCTCC

[0538] CTTCACACGGGCTCTCCTGCCGCATGTTACCAGAACACACTGGCCATAACAAAAACTGGT

[0539] TCCGCAAACACGATTAAGGGTTTCAAGCTCGGTTTGAGCAACAAGCTCTTCTGCCGCCCT

[0540] TTTCTCCTTCTCCCTCTTCTGCCTGGCGGCCTCATCCCTCTTTCTGGCAGCTTCTGCCGC

[0541] CTGATTGGCACGCAAAATCGCCATGGACTCTCGTTCGGCTTGCTCTTGGGTCTTCGTGAG

[0542] CCCTGCCTTTTTGCGGAGCTTGTAATCTCTCCATATCCGGCGTTTTTTCTCCATATCTTC

[0543] AGGAGTCATGCCCGGTCCCTCCTTCCTTTTTTGGGAGTGTACATGTCGGTCATTCCGCGT

[0544] GAGCATTCACAACAATCCGCACAGATACAGCCTTCGCATTGACCTTCCTTGGAATCTTGC

[0545] CGGGTTTCCCCGGCAAGTTCCTCGGTATAGACGTTTTTAATCGCCTCGTCCAAATAGCCG

[0546] CTTAAGTCCAGGCGTAGTTGCCTATCTTCAAAAAAATCAGAGAAAATTCCTTTGAGTGCA

[0547] GCGCGAGCTAGGAACTCTTCTTCGATTCTCGCCAACCCATCTTCCACCGCAAATTTGGCG

[0548] GTATCATACATAGATTGAGAGACAGGTATTTTCAGAGTAACAACCTTCATCACCTCATTC

[0549] CTCCTTTCATGGAAGCAAGTGTTGCGGCAAGGGAGAAGGACTTCTTGGCGCTTTCCAATA

[0550] CATGGTTGAGTAGTTCTGTCGCCATAGAATAGCGTGTCTGGTCTATGTAAACCTTTCCTT

[0551] CATCCCACTGGATACGGCCGGCATTGATGGCAGTAAACGCGGCAAGAACATCACTGTCTT

[0552] TTTCATCTATTTCCGTCAGGGCAAAAAAAGTCTTCCGCCTCTTGTCCAAAACCTTGAAAG

[0553] GAATATTCTCTTCAAATAGCTCCGCCGCATTCGCCAGAACGATGGCGGCATCCAGAAAGG

[0554] ATTCCTTCACGCTGTCGACAATCTGCTTCTGGTCGTCGTCGCAGTAATACAGGGCGTGGA

[0555] ACATATCCAGAATGGCTTCGGCCGCGCCCCGGAGCATGCAGGCATTCTCTACGAAAGCTA

[0556] CCAAATCCTCTTCATCATACCGCTTCTTCCTGGGGGCGGCGGGCTTTCGGACTGTCCTGG

[0557] CAGCCTTTGCCGGTTGACAGGGCTTCCCGTCTGTCGTACTTACTGGCTTGTGAGCAATGA

[0558] TAGGATCAGTCGTCTTGCCATTATTCACAACATCCGCCCCTTTCCCGCTGCAACGGGAGA

[0559] GGGGTTCTTCGTTTGTCAGTAGTGCTGTATTCATAGCGATCATAATCGTATTTCTTCTAT

[0560] TTTTATTTATTATCAATTAACTATTACTATTTTACCTTGTCTGTCAGAATCAATCGACGC

[0561] GCCCCGGGTGGAGGCGGGAAGGAATCATTAGCCTTCTTCGGCCAGGGAGAATCCTTCGTC

[0562] ATCCAACTCAAAAACGCCCAGCCAGGGAGCTTTGATGAAGGCGACATAGAAAATTCTCTC

[0563] ATCTTTGCGACGGACGACAATAAACTCGTCTTCCGGGGCAAAACGGTAGGTTCTGCCGCG

[0564] ATTAGGCATAGTGATGATAAGCTGGGAATTTAACATCACCCTGTCGCCCGTATCGAAAGG

[0565] CTTTGCCTGGACAGGTTCCGATGCTGCATCAATATTACTTGTAATCATTATATTATGTAT

[0566] TTTCTATTGGTTATTGCTTCCTCATGCCGTGAGGGCGGGACGGTTTTGATAAACCGTCAA

[0567] AAGCTTTCATGGGGGAGGGAGACTCCGGGCAAAACCCGGAATGCGGACTCTTGCCGGCCT

[0568] GCAGCTCGGCGTCATCCAACTCCACCGCCAGCCAGAACAGGGTTACAGCGAAGAGGCCAC

[0569] AGGCTCCACCTGCCAGGAATTGGAACAAGGTCTTCATCTACTCACTCCTCCTTCTCCATA

[0570] TTCACGGAGCAACGCCCGGCGGAACTGCTTGCCGTGTACTTTCATCTTCCCTTGCTTGCC

[0571] CCAGTACAGGATCTCGATCACATGCCCCTTGTCCTTCAACTCATGGACGGTCCGCTTGAT

[0572] CACATCCCGGTCGGAATCGTACATCAGGGCCAAGGTCTTGCAGTCGTAAAACTCTGATTC

[0573] AGGGTAGGTCATAATATTTTCATTATTAAAGTTCGTGCCAGCCGAGCCGGATAAATTCGT

[0574] CAATCAGGGCTTCTTCCATTAGGCTGCGGGCTTTTTGGGTTCAACGTTCTTCTTCGAGGT

[0575] GAGCACTCTTTGAGCTTTCGCTATTGCCAAGGCTCTCATGCGTTCTCCTATCGTTATTCC

[0576] CAATCCCCGGGCGATTCCCTCCTCTAAGAGGAGGACCGATGCGGGAAGGGAAATCCGTTC

[0577] TTCGTGTTTTTCTGTCGCGTTCATATTTCCATATTGAGAATTTTTCATCACTAGTCAATC

[0578] TATTTTTGCGGAAAGGTATCAAATATTTTACTTGCATAATGAGGATTCTTCCTCATACTC

[0579] TCCTCATGGAGAATGAAAAGGAACTCATCAAATCTTGGCTAAAGAAAAATAAAAAAAGCC

[0580] GTGAATGGTTGGCTCAGAAATGCTTTGTTAAAAAGGCGTCCGTGGATGATTGGTTTCGAT

[0581] CTAAAGGCATAATTCCTCCGGCCAAGCTTGAGCTAATCCGGAAAGAAATAAATAGGCAAG

[0582] GAGAAGAAGCGGCCAAGACAGCATTCTCTGTCCCTATATTGAATGGTGTTGCAGCGGTAG

[0583] CGGTTCCTCTTTCGGAAGATGATTTAAAACTCATTTCCAAGGCCGCACAAATCAGCGGGC

[0584] AAACTATTGAGGAGTTCATCCGTAGTGCTGCGCTGGAAGACGCGGGAGAATAATCCTCAT

[0585] TGAGGATTCTGTGGTGATGTAGGGTTGTTGTCTTCATAACTTGGATTTAGATTGAGGCAG

[0586] CAGAACTCAGGTCAAGTCCCCGTTTCCAAAAGGAAACGGGGAAATTTTTGCTGGTTGAGC

[0587] CAAATTCGAATACACGCACCAGATGTAAAATCTTAGGCAGCAGGTATATTCTCAATGGTT

[0588] AATCCCAAACGCTTCATCAACCATATTTGACCGCGGGGTGTCAAATACGGTGTCGGTTTG

[0589] GACTGCTGTGAACCGTCGGAACGCTGAATAACCCGGTAAGTGAGACGGAAATACCCAGCT

[0590] TCAATATACTCTTGGGCTGGAACATTGTGACGATTGCCGTGCCTCCCCATAATGCCTTCA

[0591] TTGCGTAGATACTCAAACAAACGGTTTTGTCCTGTCTCCATTCCATTCTGAGCGAGAATC

[0592] TTGGCAAATTCTCCAATCAGGATGCAGTCCTCGGAAACTTCCACGCATCGTCCGAAATAT

[0593] GCATAGGGAGCATCTTCCGCAGCTTTTGCTTCAAGAGCTTTTCTCTTCTCCCGTTCAGCT

[0594] TTCAATGCCTGAAGCGTCAGAATCATGGTATCCGGGTCGGCAAGCATTTCTTCCAGCTTC

[0595] TCTCCGGTGGCGTACACACCATGCTTCCGGATGGAGGGAAGGACCTCAGCGGTCACCCAC

[0596] TTCTTGAACTTCTTCGCCTCCGGCTTGCGGGAGCGGAGAATTAGAGAGTAAAGTCCGGAT

[0597] TCATTGATGATGAGAGGTGCGCGACCTCCGTTTTGAGCAATATCATTAGTAATGATGTTG

[0598] CTCTTTTCATCTTCATCCAAGTAGCTGACGGCTTGGGATACATTACCAATTTCCAGCGCA

[0599] TCGCAAACATCCTTTGCGACAAACCAGGGTTCCCCGTCCTTCACCACGGCGCGGACCGTG

[0600] CAGTTCAGGGTTTCGTTCTGGAACGGTACAACACCGTTCTGCGGCATCAAGGCATTTTCG

[0601] CCTTGCGTTTTGTGGTTGATGATATTATTAATCATTTACTTACGTTAGTTGGGCTCACGC

[0602] CCGTTATTAAAGGTCACCTGTTGGCGCAGGTGGCCTTCGTTGTTTTCGACATGGGCCGCA

[0603] TGTCGGACGAGAGGCTTACTCAAGCCACGGAAGTTCTTTTCTTGCTCCGTTCTTCCAGCC

[0604] AGGCTTTCACCTTTTCCAGATCGTAACGAGGGCGAGCTCCGCGGCGAGGTTCCGTGACTC

[0605] TGCCAATGTAGATGCAGGGCATCCCCATGCGAGCCCACATAGAAACGGTGTTTTTATGTA

[0606] CTCGGAGTGACTGTGCTAGTTCCGTAGCAGTAAGATTGCTGATGTTGCTCATATGTGTGA

[0607] TGAGATTATCACATAATCTCACATAAAGTCCACCAAAATATTTTAGCATGTGAAAAAATT

[0608] ACTCGCACATGGCTTGACATATCACATATAATCACACTATATAAATCT。

[0609] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A bacteriophage, characterized in that, The phage is Akkermansia muciniphila phage AkkZT003P, which was deposited at the Guangdong Provincial Culture Collection of Microorganisms on January 6, 2025, and its deposit number is: GDMCC NO: 65724-B1.

2. Use of the phage according to claim 1 in the preparation of a medicament for preventing and / or treating enteritis.

3. The application according to claim 2, wherein The medicament is a medicament for inhibiting Akkermansia muciniphila.

4. The application according to claim 2, wherein The medicament is a medicament for promoting the growth of Streptococcus thermophilus.

5. The application according to claim 2, wherein The medicament is a medicament for reducing the expression of intestinal pro-inflammatory cytokines.

6. The application according to claim 5, wherein The pro-inflammatory cytokines include at least one of interleukin-1, interleukin-6 and tumor necrosis factor α.

7. The application according to claim 2, wherein The medicament is a medicament for protecting the intestinal barrier.

8. The application according to claim 7, characterized in that The medicament is a medicament for increasing the expression of intestinal barrier factors.

9. The application according to claim 8, characterized in that, The barrier factors include human mucin 2.

10. A drug for preventing and / or treating enteritis, characterized in that, The medicament contains the phage according to claim 1.

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