Функциональная интерпретация единичных нуклеотидных замен в интроне 2 гена К-ras мыши, связанных с развитием рака легких
Диссертация
Впервые продемонстрировано, что горячая точка мутагенеза в протоонкогене K-ras — область 12-го ко дона — является сложным сайтом связывания белков NF-Y и GATA-6. Получены оригинальные данные о том, что под действием пульмоноканцерогенов 3-метилхолантрена (3-МХ) и нитрозоэтилмочевины (НЭМ) происходит усиление связывания факторов NF-Y и GATA-6 с районом 12 ко дона и областью полиморфизма в начале… Читать ещё >
Содержание
- 1. Введение
- 2. Обзор литературы
- 2. 1. Семейство протоонкогенов ras
- 2. 1. 1. Функциональная значимость белков семейства Ras
- 2. 1. 2. Структура Ras белков
- 2. 1. 3. Ген K-Ras и канцерогенез лёгких
- 2. 2. Единичные нуклеотидные замены
- 2. 3. Транскрипционные факторы
- 2. 3. 1. Транскрипционный фактор NF-Y
- 2. 3. 2. Транскрипционный фактор GATA
- 2. 1. Семейство протоонкогенов ras
- 3. 1. Материалы, использованные в работе
- 3. 2. Олигонуклеотиды
- 3. 3. Животные
- 3. 4. Выделение геномной ДНК из лёгких мыши
- 3. 5. Получение препаратов ДНК с использованием полимеразной цепной реакции (ПЦР)
- 3. 6. Выделение продуктов ПЦР из агарозного геля
- 3. 7. Получение экстракта ядер лёгких мышей
- 3. 8. Введение метки в ДНК с помощью фрагмента Клёнова
- 3. 9. Введение концевой метки в ДНК
- 3. 10. Метод задержки ДНК-зонда в ПААГ белками ядерного 52 экстракта
- 3. 11. Вестерн-блот анализ
- 3. 12. Поиск сайтов гиперчувствительности К ДНК-азе I
- 3. 12. 1. Обработка ядер клеток легкого ДНК-азой I
- 3. 12. 2. Гибридизация по Саузерну
- 4. 1. Изучение связывания ядерных белков с районом 58 локализации сопряженых с развитием рака легких однонуклеотидных замен в интроне 2 гена K-Ras мыши
- 4. 2. Идентификация транскрипционных факторов, 73 взаимодействующих с областью 12-ого кодона гена K-Ras мыши
- 4. 3. Влияние легочных канцерогенов 3-метилхолантрена 79 и нитрозоэтилмочевины на ДНК-связывающую активность транскрипционных факторов GATA-6 И NF-Y
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