Комплексы полинуклеотидов с белками рибосомы Escherichia coli и их свойства
Диссертация
Одним из особо сложных вопросов в этой области является вопрос о структурной и функциональной организации декодирующего участка рибосомы, т. е. участка, где взаимодействует кодон (ь мРНК с антикодоном (антикодонами) тРНК. Благодаря успехам в области иммуно-электронной микроскопии известен приблизительно тот регион, где молекула мРНК входит и выходит из 70S рибосомы Е. cot-i. Многочисленные… Читать ещё >
Содержание
- СПИСОК УПОТРЕБЛЯЕМЫХ СОКРАЩЕНИЙ
- ВВЕДЕНИЕ
- ОБЗОР ЛИТЕРАТУРЫ
- Глава I. ВЗАИМОДЕЙСТВИЕ мРНК С РИБОСОМОЙ
- 1. Связывание мРНК в инициаторном комплексе
- 2. Взаимодействие мРНК с рибосомными белками во время элонгации
- Глава II. ВЗАИМОДЕЙСТВИЕ тРНК С 30S СУБЧАСТИЦЕЙ
- Глава III. МЕХАНИЗМ ОТБОРА аа-тРНК
- ЭКСПЕРИМЕНТАЛЬНАЯ ЧАСТ
- Глава I. МАТЕРИАЛЫ И МЕТОДЫ ИССЛЕДОВАНИЯ
- 1. Использованные реактивы и материалы
- 2. Выделение суммарного белка субчастиц и рибосомы Е. со Ъъ
- 3. Выделение рибосомного белка S
- 4. Изолирование тРНК Е. ооЪг
- 5. фосфорилирование TPHKPhe и тРНК^а
- 6. Выделение препарата аминоацил-тРНК--синтетазы
- 7. Иммобилизация полинуклеотидов на гидразидагарозе
- 8. Аффинная хроматография рибосомных белков на колонках с иммобилизованными поли-нуклеотидами
- 9. Аффинная хроматография тРНК на колонках с преформированными мРНК -белковыми комплексами
- 10. Видовой анализ тРНК, связанной с мРНК -белковыми комплексами
- 11. Равновесное связывание тРНК на преформированном мРНК -белковом комплексе и определение константы диссоциации образующегося комплекса
- 12. Определение распределения мРНКИ-белкового комплекса в пределе колонки с мРНК-агарозой
- Глава II. РЕЗУЛЬТАТЫ ИССЛЕДОВАНИЯ
- 1. Результаты иммобилизации полинуклеотидов
- 2. Аффинная хроматография СБ70 на колонках с иммобилизованными полинуклеотидами
- 3. Связывание тРНК на колонках с преформированными мРНК -белковыми комплексами
- 3. 1. Связывание тРНК на мРНКИ-СБ70 комплексы
- 3. 2. Анализ тРНК, связанной на колонках с иммобилизованными полинуклеотидами без рибосомных белков
- 3. 3. Селекция деацилированной и аминоацилированной тРНК на колонках с преформированным мРНК -СБ70 комплексом
- 3. 4. Связывание деацилированной тРНК на преформированные мРНК -СБ30 и мРНКИ-СБ50 комплексы
- 4. 1. Определение константы диссоциации мРНКИ-СБ30)-тРНК комплекса
- 4. 2. Определение константы диссоциации mPHKh-S13)-tPHK комплекса
- 1. Аффинная хроматография СБ70 на колонках с иммобилизованными полинуклеотидами
- 2. Связывание тРНК на колонках с преформированными мРНКи-белковыми комплексами
- 2. 1. Связывание тРНК на мРНКи-СБ70 комплексах
- 2. 2. Связывание тРНК на мРНКи-СБ и мРНКи-СБ50 комплексах
- 2. 3. Связывание тРНК на MPHK-S4, -S9 и
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