Сравнительный анализ структуры и экспрессии генов hsp70 у видов Dropsophila с различной термальной адаптацией
Диссертация
На основании сравнения структуры генов hsp70 и паттерна БТШ70 D. virilis и D. lummei с данными, полученными для других видов Diptera, определить общее направление эволюции генов hsp70 в данной группе и их участие в термальной адаптации. Клонировать гены hsp70 D. virilis и D. lummei, определить их нуклеотидную последовательность, определить общую структуру кластера и число копий генов hsp70 D… Читать ещё >
Содержание
- Список используемых сокращений
- 1. Обзор литературы
- 1. 1. Экспрессия генов теплового шока: общие аспекты
- 1. 2. Классификация и функции БТШ
- 1. 3. Регуляция экспрессии генов ТШ
- 1. 4. Изменения в аппарате транскрипции и трансляции при ТШ
- 1. 5. Функции БТШ в апоптозе
- 1. 6. Структура и эволюция генов ТШ
- 1. 7. БТШ в адаптации организмов к неблагоприятным условиям среды обитания
- 1. 8. БТШ в медицине и экологии
- 2. Материалы и методы
- 2. 1. Материалы 51 2.1.1 Линии и виды ЭгоБорИИа
- 2. 1. 2. Штаммы Е. соИ
- 2. 1. 3. Клоны /75р70 В. melanogaster
- 2. 1. 4. Ферменты рестрикции
- 2. 1. 5. Олигонуклеотиды для анализа связывания факторов транскрипции с элементами ТШ
- 2. 1. 7. Антитела
- 2. 2. Методы
- 2. 2. 1. Условия содержания дрозофил
- 2. 2. 2. Условия теплового шока
- 2. 2. 3. Определение базальной и индуцибельной термоустойчивости
- 2. 2. 4. Включение метионина -358 в белки
- 2. 2. 5. Подсчёт общего включения метионина- 8 в белки
- 2. 2. 6. Диск-электрофорез белков с ДЦС-Иа (по Лэммли)
- 2. 2. 7. Двумерный электрофорез белков по О’Фарреллу
- 2. 2. 8. Иммуноблоттинг
- 2. 2. 9. Иммунопреципитация
- 2. 2. 10. Очистка белков методом препаративного электрофореза
- 2. 2. 11. Обработка белка модифицированным трипсином для построения пептидных карт
- 2. 2. 12. Анализ связывания факторов транскрипции с элементами ТШ
- 2. 2. 13. Включение радиоактивной метки в ДНК
- 2. 2. 14. Выделение геномной ДНК
- 2. 2. 15. Расщепление ДНК рестрицирующими эндонуклеазами
- 2. 2. 16. Электрофорез ДНК
- 2. 2. 17. Перенос и гибридизация по Саузерну
- 2. 2. 18. Определение числа копий гена по Саузерну
- 2. 2. 19. Выделение тотальной РНК
- 2. 2. 20. Электрофорез РНК
- 2. 2. 21. Нозерн-гибридизация
- 2. 2. 22. Получение геномных фаговых библиотек
- 2. 2. 23. Скрининг фаговых геномных библиотек
- 2. 2. 24. Выделение ДНК бактериофага X
- 2. 2. 25. Построение рестриктных карт рекомбинантных фагов
- 2. 2. 26. Выделение фрагментов ДНК
- 2. 2. 27. Клонирование фрагментов ДНК
- 2. 2. 28. Трансформация компетентных клеток
- 2. 2. 29. Выделение плазмидной ДНК
- 2. 2. 30. Секвенирование ДНК
- 2. 1. Материалы 51 2.1.1 Линии и виды ЭгоБорИИа
- 3. 1. Определение базальной термоустойчивости у видов и линий группы хтШ
- 3. 2. Определение индуцированной термоустойчивости у видов и линий группы irilis
- 3. 3. Определение термоустойчивости реципрокных гибридов
- 3. 4. Изучение общего белкового синтеза при ТШ у ?). irilis и И. 1итте
- 3. 5. Накопление БТШ70 при тепловом шоке
- 3. 6. Определение кинетики индукции мРНК БТШ
- 3. 7. Исследование ДНК-связывающей активности ИББ
- 3. 8. Анализ паттерна БТШ70 и динамики синтеза отдельных групп БТШ методом двумерного электрофореза
- 3. 9. Идентификация белков по молекулярным массам пептидов
- 3. 10. Рестрикционный и Саузерн-анализ генов Ыр70 В. irilis
- 3. 11. Определение числа копий гена 1гзр70 у разных линий Б. irilis и umme
- 3. 12. Определение структуры кластера генов /ир70 ?>. irilis и О. 1итте1 на основе рекомбинантных фагов X
- 3. 13. Определение нуклеотидной последовательности генов Ъзр70 и прилежащих участков 0.1итте1 и, А irilis
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