ES2652336T3 - Nuevo bacteriófago y composición antibacteriana que comprende el mismo - Google Patents

Nuevo bacteriófago y composición antibacteriana que comprende el mismo Download PDF

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ES2652336T3
ES2652336T3 ES14756932.1T ES14756932T ES2652336T3 ES 2652336 T3 ES2652336 T3 ES 2652336T3 ES 14756932 T ES14756932 T ES 14756932T ES 2652336 T3 ES2652336 T3 ES 2652336T3
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phage
enterobacteria
shigella
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ecom
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Eun Mi Shin
Gi Duk Bae
Jae Won Kim
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CJ CheilJedang Corp
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    • C12N2795/00—Bacteriophages
    • C12N2795/00011—Details
    • C12N2795/10011—Details dsDNA Bacteriophages
    • C12N2795/10211—Podoviridae
    • C12N2795/10233—Use of viral protein as therapeutic agent other than vaccine, e.g. apoptosis inducing or anti-inflammatory

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Abstract

Un bacteriófago ΦCJ20, depositado como KCCM11362P, que tiene una actividad bactericida específica contra la Escherichia coli enterotoxigénica (ETEC).

Description

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bacteriófago de la presente invención no existen. Por lo tanto, se puede confirmar que el bacteriófago era un bacteriófago aislado recientemente.
La siguiente Tabla 2 muestra los resultados obtenidos comparando homólogos de la secuencia genómica del ΦCJ20
5 y se muestran posteriormente secuencias descodificadas del genoma de otros bacteriófagos, y la secuencia genómica parcial del ΦCJ20. Tabla 2
[Tabla 2]
Consulta
Sujeto Valoración Identidades
Nombre
Longitud Inicio Final Descripción E-Valor Coincidencia /Total Pct. (%)
contig00001_orf00002
354 1 351 proteína hipotética conservada PseT.3 [fago de Enterobacterias vB_EcoM-VR7] 5E-42 110/117 94
contig00001_orf00004
489 7 486 VR7_gp239 hipotética conservada [Fago de Enterobacterias vBEcoM-VR7] 2E-82 152/160 95
contig00001_orf00038
933 1 930 Iniciador de la placa base del tubo de cola [fago de Shigella SP18] 4E-172 309/310 99
contig00001_orf00047
465 1 462 Proteína de unión a ssADN y reparación de recombinación UvsY [fago de Shigella SP18] 1E-82 152/154 98
contig00001_orf00072
492 1 489 Proteína terminasa pequeña [fago de Enterobacterias vB_EcoM-VR7] 1E-77 162/163 99
contig00001_orf00074
801 1 798 Proteína gp15 de terminación y estabilizadora de la vaina de la cola [fago de Enterobacterias vB_EcoM-VR7] 1E-142 264/266 99
contig00001_orf00066
261 1 135 Proteína del núcleo de procápside precursora de péptidos internos [fago de Shigella SP18] 1E-16 45/45 100
contig00001_orf00076
939 1 936 Proteína gp13 del cuello [fago de Enterobacterias vB_EcoM-VR7] 3E-169 308/312 98
contig00001_orf00093
456 1 453 proteína hipotética conservada 57B [fago de Enterobacterias vB_EcoM-VR7] 3E-82 150/151 99
11
Consulta
Sujeto Valoración Identidades
Nombre
Longitud Inicio Final Descripción E-Valor Coincidencia /Total Pct. (%)
contig00001_orf00087
603 1 600 Subunidad en cuña gp53 de la placa base [fago de Enterobacterias vB_EcoM-VR7] 2E-112 192/200 96
contig00002_orf00005
456 1 453 lisozima [fago de Enterobacterias T7] 4E-81 143/151 94
contig00002_orf00007
699 1 633 gp2.5 [fago de Enterobacterias 13a] 4E-105 208/211 98
contig00002_orf00011
309 1 300 gp1.7 [fago de Enterobacterias 285P] 1E-47 92/102 90
contig00002_orf00035
1032 1 996 gp10A [fago de Enterobacterias 13a] 1E-137 313/333 93
contig00003_orf00016
363 1 360 SP18gp253 hipotética conservada [fago de Shigella SP18] 3E-62 117/120 97
contig00001_orf00056
1008 1 1005 ARN ligasa 2 [fago de Shigella SP18] 6E-172 321/335 95
contig00001_orf00101
390 10 387 Proteína de fago hipotética [Escherichia phage wV7] 6E-66 120/126 95
contig00001_orf00089
825 1 822 Proteína gp2 protectora final de ADN [fago de Enterobacterias vB_EcoM-VR7] 7E-147 273/274 99
contig00001_orf00019
348 1 345 Cochaperona de ensamblaje de cápside con GroEL [fago de Shigella SP18] 2E-61 115/115 100
contig00001_orf00008
906 1 903 polinucleótido 5’-cinasa y 3’-fosfatasa [fago de Shigella SP18] 1E-170 287/301 95
contig00001_orf00006
321 1 318 EpJS10_0215 hipotética conservada [fago de Enterobacterias JS10] 2E-50 102/106 96
contig00001_orf00020
324 1 321 Proteína accesoria de inhibición del fenotipo de lisis rápida [fago de Shigella SP18] 5E-53 105/107 98
12
Consulta
Sujeto Valoración Identidades
Nombre
Longitud Inicio Final Descripción E-Valor Coincidencia /Total Pct. (%)
contig00001_orf00017
582 1 579 Desoxicitidilato desaminasa [fago de Enterobacterias vB_EcoM-VR7] 6E-111 191/193 98
contig00001_orf00081
870 1 867 Conector de fibra de la cola de cuña de placa base [fago de Shigella SP18] 5E-161 286/289 98
contig00001_orf00013
360 1 357 SP18gp231 hipotética conservada [fago de Shigella SP18] 2E-47 113/119 94
contig00001_orf00037
324 1 321 SP18gp210 hipotética conservada [fago de Shigella SP18] 3E-51 104/107 97
contig00001_orf00079
666 1 663 Subunidad cuña de la placa base y punta de la cola[fago de Shigella SP18] 2E-118 215/221 97
contig00002_orf00043
903 1 900 exonucleasa [fago de Enterobacterias T7] 6E-168 293/300 97
contig00003_orf00017
912 7 876 Proteína de unión al ADN de cadena sencilla [fago de Shigella SP18] 6E-145 287/290 98
contig00001_orf00063
819 1 816 Proteína central de procápside [fago de Enterobacterias vB_EcoM-VR7] 5E-99 267/272 98
contig00001_orf00044
729 1 726 Ensamble catalítico del núcleo de la placa base [fago de Shigella SP18] 2E-128 241/242 99
contig00001_orf00088
354 1 351 Proteína de terminación de la cápside [fago de Shigella SP18] 2E-64 115/117 98
contig00001_orf00023
369 1 366 SP18gp221 hipotética conservada [fago de Shigella SP18] 4E-68 122/122 100
contig00001_orf00046
390 1 387 Subunidad en cuña de la placa base [fago de Enterobacterias vB_EcoM-VR7] 5E-66 128/129 99
contig00001_orf00053
849 1 846 Proteína de la cápside externa [fago de Shigella SP18] 3E-141 268/282 95
13
Consulta
Sujeto Valoración Identidades
Nombre
Longitud Inicio Final Descripción E-Valor Coincidencia /Total Pct. (%)
contig00001_orf00042
459 1 456 Subunidad gp28 del núcleo distal de la placa base [fago de Enterobacterias vB_EcoM-VR7] 3E-83 150/152 98
contig00001_orf00064
639 1 636 Proteína y proteasa del núcleo de matriz de procápside [fago de Shigella SP18] 4E-118 210/212 99
contig00001_orf00052
669 1 666 Inhibidor de la proteasa gp21 de procápside [fago de Shigella SP18] 2E-91 206/222 92
contig00001_orf00045
624 1 621 Subunidad del núcleo de placa base [fago de Shigella SP18] 4E-115 206/207 99
contig00001_orf00086
178 8 1 178 5 Subunidad central de la placa base y lisozima de la cola [fago de Shigella SP18] 0 588/595 98
contig00001_orf00094
366 1 363 VR7_gp156 hipotética conservada [fago de Enterobacterias vB_EcoM-VR7] 2E-48 119/121 98
contig00001_orf00016
336 1 333 SP18gp229 hipotética conservada [fago de Shigella SP18] 2E-54 108/111 97
contig00002_orf00001
408 1 405 proteína 4.7 [fago de Enterobacterias T7] 2E-54 123/135 91
contig00002_orf00013
102 3 1 102 0 ADN ligasa [fago de Enterobacterias T7] 0 317/359 88
contig00002_orf00027
150 0 1 975 gp17 [fago de Enterobacterias 13a] 6E-120 268/330 81
contig00002_orf00033
591 1 588 Proteína de la cola [fago de Yersinia pestis phiA1122] 2E-99 194/196 98
contig00003_orf00003
411 1 408 endonucleasa II [fago de Shigella SP18] 5E-72 136/136 100
contig00003_orf00008
861 1 858 dTMP timidilato sintasa [fago de Enterobacterias vB_EcoM-VR7] 2E-146 245/286 85
contig00003_orf00019
669 1 648 Cargador de gp41 ADN helicasa [fago de Shigella SP18] 4E-118 211/216 97
14
Consulta
Sujeto Valoración Identidades
Nombre
Longitud Inicio Final Descripción E-Valor Coincidencia /Total Pct. (%)
contig00001_orf00110
306 1 303 SP18gp146 hipotética conservada [fago de Shigella SP18] 4E-50 99/101 98
contig00001_orf00058
128 1 1 127 8 Proteína vertex de la cápside [fago de Enterobacterias vB_EcoM-VR7] 0 411/426 96
contig00001_orf00041
1746 1 1743 Subunidad del núcleo de la placa base determinante de la longitud de la cola [fago de Shigella SP18] 0 570/581 98
contig00001_orf00084
1959 1 1956 Subunidad en cuña de la placa base [fago de Shigella SP18] 0 648/652 99
contig00001_orf00080
1785 1 1782 Subunidad gp10 en cuña de la placa base y punta de la cola [fago de Enterobacterias vB_EcoM-VR7] 0 583/594 98
contig00001_orf00040
1095 1 1092 Protección del tubo de cola de la placa base [fago de Shigella SP18] 0 362/364 99
contig00001_orf00090
600 1 597 Proteína gp3 de terminación de la cola y estabilizador de la vaina [fago de Enterobacterias vB_EcoM-VR7] 2E-112 199/199 100
contig00001_orf00068
492 1 489 Proteína del tubo de la cola [fago de Shigella SP18] 2E-93 161/163 98
contig00001_orf00091
678 1 675 gp1 desoxinucleósido monofosfato cinase [fago de Enterobacterias vB_EcoM-VR7] 2E-127 222/225 98
contig00001_orf00032
717 1 714 SP18gp215 hipotética conservada [fago de Shigella SP18] 3E-128 235/238 98
contig00001_orf00025
366 1 363 SP18gp220 hipotética conservada [fago de Shigella SP18] 1E-66 120/121 99
contig00001_orf00001
405 1 402 Inhibidor de transcripción de huésped [fago de Shigella SP18] 2E-61 131/134 97
15
Consulta
Sujeto Valoración Identidades
Nombre
Longitud Inicio Final Descripción E-Valor Coincidencia /Total Pct. (%)
contig00001_orf00112
594 1 591 SP18gp145 hipotética conservada [fago de Shigella SP18] 7E-100 193/197 97
contig00001_orf00075
786 1 753 Proteína del cuello [fago de Enterobacterias vB_EcoM-VR7] 1E-144 251/251 100
contig00002_orf00006
456 1 453 gp3 [fago de Enterobacterias 13a] 1E-82 149/151 98
contig00002_orf00017
1080 1 1077 Proteína cinasa [fago de Enterobacterias T7] 2E-163 325/359 90
contig00002_orf00024
360 1 357 gp18.5 [fago de Enterobacterias 13a] 1E-57 112/119 94
contig00002_orf00030
591 1 588 Proteína interna del virión [fago de Yersinia pestis phiA1122] 1E-74 178/196 90
contig00003_orf00009
582 1 579 Frd dihidrofolato reductasa [fago de Enterobacterias vB_EcoM-VR7] 6E-105 187/193 96
contig00003_orf00011
342 1 339 SP18gp249 hipotética conservada [fago de Shigella SP18] 3E-59 111/113 98
contig00003_orf00014
366 1 363 SP18gp252 hipotética conservada [fago de Shigella SP18] 2E-60 113/121 93
Véase por favor la [secuencia del genoma parcial de ΦCJ20] adjunta.
Ejemplo 7
5 Ensayo de estabilidad de ΦCJ20 dependiendo del pH
Con el fin de confirmar si el bacteriófago ΦCJ20 puede tener estabilidad o no en un ambiente a pH bajo en el estómago, se llevó a cabo un ensayo de estabilidad en un amplio intervalo de pH (pH 3,0, 3,5, 4,0, 5,5, 6,4, 7,5, 8,3,
10 9,2, y 11,0).
Para el ensayo, se prepararon soluciones a distintos pH (tampón de acetato sódico (pH 4,0, pH 5,5, y pH 6,4), tampón de citrato sódico (pH 3,0 y pH 3,5), tampón de fosfato sódico (pH 6,9 y pH 7,4), y solución Tris-HCl (pH 8,2, pH 9,0, pH 9,8, y pH 11,0)) a una concentración de 0,2 M, respectivamente.
15 Después, 180 µl de cada una de las soluciones de pH se mezclaron con 20 µl de solución de bacteriófago que tiene un título de 2,1 x 1010 ufp/ml de manera que una concentración de cada pH era de 1 M, y cada una de las soluciones de pH se dejó a temperatura ambiente durante 2 horas. En un grupo de control, 20 µl de solución de bacteriófago (2,1 x 1010 ufp/ml) se mezcló con 180 µl de solución SM y entonces se dejó a temperatura ambiente durante 2 horas.
20 Entonces, la reacción de solución se diluyó poco a poco, 10 µl de la solución diluida en cada paso se depositó y se cultivó a 30 ⁰C durante 18 horas por un método de depósito en agar blando, y el título se midió mediante la presencia o ausencia de lisis (FIG. 4).
16
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Claims (1)

  1. imagen1
ES14756932.1T 2013-02-27 2014-02-24 Nuevo bacteriófago y composición antibacteriana que comprende el mismo Active ES2652336T3 (es)

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KR1020130021498A KR101381796B1 (ko) 2013-02-27 2013-02-27 신규 박테리오파지 및 이를 포함하는 항균 조성물
KR20130021498 2013-02-27
PCT/KR2014/001476 WO2014133289A1 (en) 2013-02-27 2014-02-24 Novel bacteriophage and antibacterial composition comprising the same

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KR101381795B1 (ko) * 2013-02-27 2014-04-07 씨제이제일제당 (주) 신규 박테리오파지 및 이를 포함하는 항균 조성물
KR101381793B1 (ko) * 2013-02-27 2014-04-07 씨제이제일제당 (주) 신규 박테리오파지 및 이를 포함하는 항균 조성물
KR101592177B1 (ko) 2014-05-07 2016-02-05 주식회사 인트론바이오테크놀로지 대장균에 대한 광범위 항균 활성을 갖는 박테리오파지를 활용한 대장균 감염을 방지 및 처치하는 방법
WO2016168560A1 (en) 2015-04-16 2016-10-20 Kennesaw State University Research And Service Foundation, Inc. Escherichia coli o157:h7 bacteriophage φ241
KR101993125B1 (ko) * 2016-12-22 2019-06-26 주식회사 옵티팜 신규한 esbl 생성 대장균 특이 박테리오파지 eco4 및 이를 포함하는 항균 조성물
CN109207435A (zh) * 2017-07-04 2019-01-15 中国科学院大学 一株新型大肠杆菌h21噬菌体st31的分离鉴定
CN111057681B (zh) * 2019-12-02 2023-02-28 衡阳师范学院 一株噬菌体及其应用
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