RU2326170C2 - Corynebacterium glutamicum genes encoding proteins of the phosphoenolpyruvate-sugar-phosphotransferase system - Google Patents

Corynebacterium glutamicum genes encoding proteins of the phosphoenolpyruvate-sugar-phosphotransferase system Download PDF

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RU2326170C2
RU2326170C2 RU2002102452/13A RU2002102452A RU2326170C2 RU 2326170 C2 RU2326170 C2 RU 2326170C2 RU 2002102452/13 A RU2002102452/13 A RU 2002102452/13A RU 2002102452 A RU2002102452 A RU 2002102452A RU 2326170 C2 RU2326170 C2 RU 2326170C2
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nucleic acid
acid molecule
polypeptide
seq
activity
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RU2002102452A (en
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Маркус ПОМПЕЙУС (DE)
Маркус ПОМПЕЙУС
Буркхард КРЕГЕР (DE)
Буркхард КРЕГЕР
Хартвиг ШРЕДЕР (DE)
Хартвиг ШРЕДЕР
Оскар ЦЕЛЬДЕР (DE)
Оскар Цельдер
Грегор ХАБЕРХАУЕР (DE)
Грегор ХАБЕРХАУЕР
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Басф Акциенгезелльшафт
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract

FIELD: biotechnologies.
SUBSTANCE: A description is given of extracted molecules of the Corynebacterium glutamicum nucleic acids that encode polypeptides having activity of the II ABC saccharose-specific component. The invention relates as well to recombinant expression vectors comprising the described nucleic acids molecules. A method for producing a host cell containing the described expression vectors is exposed therein. The present invention relates as well to a method for producing the described polypeptide by means of cultivating said host cell. A method of testing patients for Corynebacterium glutamicum comprising the detection of the described nucleic acids molecules is exposed therein.
EFFECT: diversification of the nucleic acids molecules encoding proteins of the phosphoenolpyruvate- sugar-phosphotransferase system.
27 cl, 4 dwg, 13 tbl

Description

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Claims (27)

1. Выделенная молекула нуклеиновой кислоты Corynebacterium glutamicum, содержащая нуклеотидную последовательность SEQ ID NO:1, или комплементарная ей последовательность, где указанная молекула кодирует полипептид, обладающий активностью сахароза-специфического компонента II ABC.1. An isolated Corynebacterium glutamicum nucleic acid molecule containing the nucleotide sequence of SEQ ID NO: 1, or a sequence complementary to it, wherein said molecule encodes a polypeptide having the activity of a sucrose-specific component II ABC. 2. Выделенная молекула нуклеиновой кислоты, которая кодирует полипептид, содержащий аминокислотную последовательность SEQ ID NO:2 и обладающий активностью сахароза-специфического компонента II ABC, или комплементарная ей последовательность.2. The selected nucleic acid molecule that encodes a polypeptide containing the amino acid sequence of SEQ ID NO: 2 and having the activity of a sucrose-specific component II ABC, or a sequence complementary to it. 3. Выделенная молекула нуклеиновой кислоты, которая кодирует природный аллельный вариант полипептида, содержащего аминокислотную последовательность SEQ ID NO:2 и обладающего активностью сахароза-специфического компонента II ABC, или комплементарная ей последовательность.3. An isolated nucleic acid molecule that encodes a natural allelic variant of a polypeptide containing the amino acid sequence of SEQ ID NO: 2 and having the activity of a sucrose-specific component II ABC, or a sequence complementary to it. 4. Выделенная молекула нуклеиновой кислоты, содержащая нуклеотидную последовательность, которая по меньшей мере на 50% идентична полноразмерной нуклеотидной последовательности SEQ ID NO:1, или комплементарная ей последовательность, где указанная молекула нуклеиновой кислоты кодирует полипептид, обладающий активностью сахароза-специфического компонента II ABC.4. An isolated nucleic acid molecule containing a nucleotide sequence that is at least 50% identical to the full-length nucleotide sequence of SEQ ID NO: 1, or a sequence complementary to it, wherein said nucleic acid molecule encodes a polypeptide having the activity of a sucrose-specific component II ABC. 5. Выделенная молекула нуклеиновой кислоты, содержащая фрагмент из по меньшей мере 15 последовательно расположенных нуклеотидов нуклеотидной последовательности SEQ ID NO:1, или комплементарная ей последовательность, где указанная молекула кодирует полипептид, обладающий активностью сахароза-специфического компонента II ABC.5. An isolated nucleic acid molecule containing a fragment of at least 15 consecutive nucleotides of the nucleotide sequence of SEQ ID NO: 1, or a sequence complementary to it, wherein said molecule encodes a polypeptide having the activity of a sucrose-specific component II ABC. 6. Выделенная молекула нуклеиновой кислоты, которая в жестких условиях гибридизуется с молекулой нуклеиновой кислоты по любому из пп.1-5.6. The selected nucleic acid molecule, which under stringent conditions, hybridizes with the nucleic acid molecule according to any one of claims 1 to 5. 7. Выделенная молекула нуклеиновой кислоты, содержащая молекулу нуклеиновой кислоты по любому из пп.1-6 и нуклеотидную последовательность, кодирующую гетерологичный полипептид.7. The selected nucleic acid molecule containing the nucleic acid molecule according to any one of claims 1 to 6 and a nucleotide sequence encoding a heterologous polypeptide. 8. Рекомбинантный вектор экспрессии, содержащий молекулу нуклеиновой кислоты по любому из пп.1-7 и одну или несколько регуляторных последовательностей.8. A recombinant expression vector comprising a nucleic acid molecule according to any one of claims 1 to 7 and one or more regulatory sequences. 9. Рекомбинантный вектор по п.8, который является плазмидой.9. The recombinant vector of claim 8, which is a plasmid. 10. Способ получения рекомбинантной клетки-хозяина, включающий трансфецирование клетки-хозяина вектором экспрессии по пп.8-9.10. A method of obtaining a recombinant host cell, comprising transfection of the host cell with the expression vector according to claims 8-9. 11. Способ по п.10, где указанная клетка является микроорганизмом.11. The method of claim 10, wherein said cell is a microorganism. 12. Способ по п.11, где указанная клетка принадлежит роду Corynebacterium или Brevibacterium.12. The method according to claim 11, where the specified cell belongs to the genus Corynebacterium or Brevibacterium. 13. Способ получения рекомбинантной клетки-хозяина, включающий трансфецирование клетки-хозяина молекулой нуклеиновой кислоты, содержащей нуклеотидную последовательность SEQ ID NO:1, где молекула нуклеиновой кислоты имеет одну или несколько модификаций нуклеиновой кислоты по сравнению с последовательностью, представленной в SEQ ID NO:1, и где молекула нуклеиновой кислоты кодирует полипептид, обладающий активностью сахароза-специфического компонента II ABC.13. A method of producing a recombinant host cell, comprising transfecting the host cell with a nucleic acid molecule containing the nucleotide sequence of SEQ ID NO: 1, where the nucleic acid molecule has one or more modifications of the nucleic acid compared to the sequence shown in SEQ ID NO: 1 , and where the nucleic acid molecule encodes a polypeptide having the activity of a sucrose-specific component II ABC. 14. Способ получения рекомбинантной клетки-хозяина, включающий трансфецирование клетки-хозяина молекулой нуклеиновой кислоты, содержащей нуклеотидную последовательность SEQ ID NO:1, где регуляторная область молекулы нуклеиновой кислоты модифицирована относительно регуляторной области этой молекулы дикого типа, и где молекула нуклеиновой кислоты кодирует полипептид, обладающий активностью сахароза-специфического компонента II ABC.14. A method of producing a recombinant host cell, comprising transfecting the host cell with a nucleic acid molecule containing the nucleotide sequence of SEQ ID NO: 1, where the regulatory region of the nucleic acid molecule is modified relative to the regulatory region of this wild-type molecule, and where the nucleic acid molecule encodes a polypeptide, possessing the activity of a sucrose-specific component II ABC. 15. Способ получения полипептида, включающий культивирование рекомбинантной клетки-хозяина, трансфецированной вектором по п.8, в подходящей культуральной среде с получением, таким образом, полипептида, обладающего активностью сахароза-специфического компонента II ABC.15. A method for producing a polypeptide, comprising culturing a recombinant host cell transfected with the vector of claim 8 in a suitable culture medium, thereby producing a polypeptide having the activity of a sucrose-specific component II ABC. 16. Выделенный полипептид, содержащий аминокислотную последовательность SEQ ID NO:2, где указанный полипептид обладает активностью сахароза-специфического компонента II ABC.16. The selected polypeptide containing the amino acid sequence of SEQ ID NO: 2, where the specified polypeptide has the activity of a sucrose-specific component II ABC. 17. Выделенный полипептид, содержащий природный аллельный вариант полипептида, содержащего указанную аминокислотную последовательность SEQ ID NO:2, где указанный природный аллельный вариант обладает активностью сахароза-специфического компонента II ABC.17. The selected polypeptide containing a natural allelic variant of the polypeptide containing the specified amino acid sequence of SEQ ID NO: 2, where the specified natural allelic variant has the activity of a sucrose-specific component II ABC. 18. Выделенный полипептид, который кодируется молекулой нуклеиновой кислоты, содержащей нуклеотидную последовательность, которая по меньшей мере на 50% идентична полноразмерной последовательности нуклеиновой кислоты SEQ ID NO:1, где указанный полипептид обладает активностью сахароза-специфического компонента II ABC.18. The selected polypeptide, which is encoded by a nucleic acid molecule containing a nucleotide sequence that is at least 50% identical to the full-length nucleic acid sequence of SEQ ID NO: 1, where the specified polypeptide has the activity of a sucrose-specific component II ABC. 19. Выделенный полипептид, содержащий аминокислотную последовательность, которая по меньшей мере на 50% идентична полноразмерной аминокислотной последовательности SEQ ID NO:2, где указанный полипептид обладает активностью сахароза-специфического компонента II ABC.19. An isolated polypeptide comprising an amino acid sequence that is at least 50% identical to the full length amino acid sequence of SEQ ID NO: 2, wherein said polypeptide has the activity of a sucrose-specific component II ABC. 20. Способ по п.12, где указанная клетка выбрана из группы, включающей Corynebacterium glutamicum, Corynebacterium herculis, Corynebacterium, lilium, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium acetophilum, Corynebacterium ammoniagenes, Corynebacterium fujiokense, Corynebacterium nitrilophilus, Brevibacterium ammoniagenes, Brevibacterium butanicum, Brevibacterium divaricatum, Brevibacterium flavum, Brevibacterium healii, Brevibacterium ketoglutamicum, Brevibacterium ketosoreductum, Brevibacterium lactofermentum, Brevibacterium linens, Brevibacterium paraffinolyticum и штаммы, представленные в таблице 3.20. The method of claim 12, wherein said cell is selected from the group consisting of Corynebacterium glutamicum, Corynebacterium herculis, Corynebacterium, lilium, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium acetophilum, Corynebacterium ammoniagenes, Corynebacterium fujiokense, Corynebacterium nitrilophilus, Brevibacterium ammoniagenes, Brevibacterium butanicum, Brevibacterium divaricatum, Brevibacterium flavum, Brevibacterium healii, Brevibacterium ketoglutamicum, Brevibacterium ketosoreductum, Brevibacterium lactofermentum, Brevibacterium linens, Brevibacterium paraffinolyticum and the strains shown in Table 3. 21. Способ диагностики Corynebacterium diphtheriae у пациента, включающий обнаружение по меньшей мере одной молекулы нуклеиновой кислоты по пп.1-5 или по меньшей мере одной молекулы полипептида по пп.16-19, 22 или 24, то есть, проведение диагностики или определение активности Corynebacterium diphtheriae у пациента.21. A method for diagnosing Corynebacterium diphtheriae in a patient, comprising detecting at least one nucleic acid molecule according to claims 1-5 or at least one polypeptide molecule according to claims 16-19, 22 or 24, that is, conducting a diagnosis or determining activity Corynebacterium diphtheriae in a patient. 22. Выделенный полипептид, содержащий фрагмент аминокислотной последовательности SEQ ID NO:2, где указанный фрагмент сохраняет активность сахароза-специфического компонента II ABC.22. The selected polypeptide containing a fragment of the amino acid sequence of SEQ ID NO: 2, where the specified fragment retains the activity of the sucrose-specific component II ABC. 23. Выделенная молекула нуклеиновой кислоты, содержащая фрагмент по меньшей мере из 15 последовательно расположенных нуклеотидов нуклеотидной последовательности SEQ ID NO:1, или комплементарная ей последовательность, где указанная молекула нуклеиновой кислоты может использоваться в качестве зонда.23. An isolated nucleic acid molecule containing a fragment of at least 15 nucleotides of the nucleotide sequence of SEQ ID NO: 1, or a sequence complementary to it, wherein said nucleic acid molecule can be used as a probe. 24. Выделенная молекула нуклеиновой кислоты, содержащая нуклеиновую последовательность SEQ ID NO:1, где указанный полипептид обладает активностью сахароза-специфического компонента II ABC.24. An isolated nucleic acid molecule containing the nucleic acid sequence of SEQ ID NO: 1, wherein said polypeptide has the activity of a sucrose-specific component II ABC. 25. Выделенная молекула нуклеиновой кислоты, содержащая фрагмент по меньшей мере из 15 последовательно расположенных нуклеотидов нуклеотидной последовательности SEQ ID NO:1, или комплементарная ей последовательность, где указанная молекула нуклеиновой кислоты может использоваться в качестве антисмысловой молекулы.25. An isolated nucleic acid molecule containing a fragment of at least 15 nucleotides of the nucleotide sequence of SEQ ID NO: 1, or a sequence complementary to it, wherein said nucleic acid molecule can be used as an antisense molecule. 26. Выделенная молекула нуклеиновой кислоты, содержащая фрагмент по меньшей мере из 15 последовательно расположенных нуклеотидов нуклеотидной последовательности SEQ ID NO:1, или комплементарная ей последовательность, где указанная молекула нуклеиновой кислоты может использоваться в качестве праймера.26. An isolated nucleic acid molecule containing a fragment of at least 15 consecutive nucleotides of the nucleotide sequence of SEQ ID NO: 1, or a complementary sequence to it, where the specified nucleic acid molecule can be used as a primer. 27. Выделенный полипептид по любому из пп.16-19, 22 или 24 дополнительно содержащий гетерологичные аминокислотные последовательности.27. The selected polypeptide according to any one of claims 16-19, 22 or 24 further comprising heterologous amino acid sequences.
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