Свойства и роль пирофосфат-зависимых 6-фосфофруктокиназ у аэробных метанотрофов и метилобактерий
Диссертация
Апробация работы. Основные положения диссертации доложены на ежегодных отчетных конференциях ИБФМ РАН (2006;201 Orr), на Всероссийской молодежной школе-конференции «Актуальные аспекты современной микробиологии» (Москва, 2007), на международной научной конференции «Современное состояние и перспективы развития микробиологии и биотехнологии» (Минск, 2008), на I и III школе-конференции «Биология… Читать ещё >
Содержание
- Список сокращений
- ОБЗОР ЛИТЕРАТУРЫ
- Глава 1. Пирофосфатзависимая 6-фосфофруктокиназа
- 1. 1. Свойства и метаболическая роль ПФК
- 1. 2. Характеристика ПФК метилотрофных бактерий
- 1. 3. Структурный анализ ПФК
- 1. 4. Классификация и филогенетический анализ ФФК
- 1. 5. Фруктозо-1,6-бисфосфатальдолаза
- Глава 2. Характеристика аэробных метилотрофных бактерий
- 2. 1. Энергетический метаболизм
- 2. 2. Конструктивный метаболизм
- ЭКСПЕРИМЕНТАЛЬНАЯ ЧАСТ
- Глава 3. Материалы и методы исследования
- 3. 1. Культивирование бактерий
- 3. 2. Выделение геномной ДНК
- 3. 3. Выделение плазмидной ДНК
- 3. 4. Выделение РНК
- 3. 5. Определение стартовой точки транскрипции методом удлинения праймера
- 3. 6. ОТ-ПЦР анализ
- 3. 7. Нозерн-блот анализ
- 3. 8. Получение компетентных клеток и их трансформация
- 3. 9. Клонирование и экспрессия геновр/р я/Ьа
- 3. 10. Выделение рекомбинантных белков аффинной металлхелатной хроматографией
- 3. 11. Измерение активности ПФК
- 3. 12. Измерение активности ФБА
- 3. 13. Определение субъединичного состава белков и молекулярной массы субъединиц
- 3. 14. Электрофорез в градиенте пористости ПААГ
- 3. 15. Определение нативной массы белка с помощью гель-фильтрации
- 3. 16. Анализ нуклеотидных и аминокислотных последовательностей
- 3. 17. Аналитические методы
- РЕЗУЛЬТАТЫ И ОБСУЖДЕНИЕ
- Глава 4. Характеристикарекомбинантных ПФК и ФБА Methylococcus capsulatus Bath
- 4. 1. Клонирование гена pfp и очистка ПФК-HiSô
- 4. 2. Характеристика рекомбинантной ПФК
- 4. 3. Изучение транскрипционной организации pfp и hpp генов
- 4. 4. Клонирование гена fba и очистка ФБА-Шэб
- 4. 5. Характеристика рекомбинантной ФБА
- 4. 6. Изучение транскрипции гена fba
- 4. 7. Филогенетический анализ ФБА
- 4. 8. Роль ПФК и ФБА в метаболизме Methylococcus capsulatus Bath
- Глава 5. Характеристика рекомбинантной ПФК Methylomicrobium alcaliphilum 20Z
- 5. 1. Клонирование гена pfp и очистка ПФК-HiSô
- 5. 2. Характеристика рекомбинантной ПФК
- 5. 3. Роль ПФК в метаболизме Methylomicrobium alcaliphilum 20Z
- Глава 6. Характеристика рекомбинантных ПФК Methylosinus trichosporium ОВЗЬ и Methylobacterium nodularis ORS
- 6. 1. Клонирование генов pfp и очистка рекомбинантных ПФК
- 6. 2. Характеристика рекомбинантных ПФК
- 6. 3. Роль ПФК в метаболизме Methylosinus trichosporium ОВЗЬ и Methylobacterium nodularis ORS
- Глава 7. Филогенетический анализ ПФК метилотрофных бактерий
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