Пространственно-временная организация репликации субхромосомных доменов ДНК в ядрах клеток человека
Диссертация
Решение задачи осоответствии отдельных наблюдаемых на препаратах ядер PC какому-либо определенному периоду S-фазы может быть облегчено, если использовать методы синхронизации клеточных популяций. При этом, однако, нужно учитывать, что в любой синхронизированной клеточной культуре всегда присутствуют клетки, которые могут находиться в любой. точке S-фазы. Процедура синхронизации обогащает клетками… Читать ещё >
Содержание
- I. ВВЕДЕНИЕ.б
- II. ОБЗОР ЛИТЕРАТУРЫ
- 1. Временная структура S-фазы клеточного цикла эукариот
- 2. Скорость репликации ДНК
- 3. Характеристики элементарных единиц репликации ДНК
- 3. 1. Размер и расположение на молекуле ДНК
- 3. 2. Функционально-структурные характеристики
- 4. Организация хроматина в интерфазном ядре
- 5. Фокальные центры репликации ДНК в ядрах клеток млекопитающих
- 5. 1. Д искретность мест репликативного синтеза ДНК
- 5. 2. Дискретное распределение белков, участвующих в репликации
- 5. 3. Пространственно временные характеристики РФ
- III. МАТЕРИАЛЫ И МЕТОДЫ
- 1. Ведение культур клеток
- 2. Синхронизация клеточной культуры
- 3. Проточная ДНК цитометрия
- 4. Включение в клетки репликативной метки
- 5. Приготовление препаратов ядер после включения репликативной метки
- 6. Приготовление препаратов нитей ДНК с одиночной репликативной меткой, расправленных на стекле
- 7. Денатурация ДНК на препаратах
- 8. Иммунофлуоресцентное окрашивание препаратов ядер включивших репликативную метку
- 9. Микроскопия и анализ изображений
- 9. 1. Широкопольная флуоресцентная микроскопия
- 9. 2. Анализ данных
- 9. 3. Конфокальная флуоресцентная микроскопия
- IV. РЕЗУЛЬТАТЫ И ОБСУЖДЕНИЕ Часть 1. Исследование динамики прохождения клетками К562 S-фазы клеточного цикла методом проточной ДНК-цитометрии
- Часть 2. Исследование препаратов ядер включивших репликативную метку
- 1. Дифракционная природа наблюдаемых фокусов репликации
- 2. Определение толщины ядер на препаратах
- 3. Влияние процедуры приготовления препаратов на наблюдаемую картину распределения мини-фокусов репликации в ядре
- 4. Динамика изменения мини-фокусов репликации при увеличении продолжительности мечения
- 5. Гетерогенность фокусов репликации
- 6. Определение содержания ДНК в различных мини-фокусах
- 7. Зависимость картин распределения РФ от положения клетки в S-фазе
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