Молекулярно-цитогенетический анализ ключевых событий мейоза у ржи Secale cereale L
Диссертация
Продукт гена SY9 участвует в сборке линейных трактов белка Asyl. Мутация sy9 нарушает этот процесс, но не сопровождается нарушениями ни в формировании «раннего кластера», ни в образовании промежуточных продуктов рекомбинации, связанных с участием рекомбиногенных белков Rad51/Dmcl. С другой стороны, дефект «раннего кластера» и сопряженный с этим блок рекомбинации на этапе взаимодействия ДНК… Читать ещё >
Содержание
- СОКРАЩЕНИЯ
- Глава 1. СОВРЕМЕННЫЕ ПРЕДСТАВЛЕНИЯ О МЕЙОЗЕ ОБЗОР ЛИТЕРАТУРЫ).14'
- 1. 1. Основные события мейоза
- 1. 2. Ключевые события профазы I мейоза!
- 1. 2. 1. Передислокации доменов хромосом, кластеризация центромеров и теломеров
- 1. 2. 2. Гомологичное спаривание хромосом и сборка синаптонемного комплекса
- 1. 2. 2. 1. Белки поперечных филаментов СК
- 1. 2. 2. 2. Белки осевых элементов гомологичных хромосом и латеральных элементов СК, не относящиеся к когезину
- 1. 2. 3. Рекомбинация гомологичных хромосом
- 1. 2. 3. 1. Два пути осуществления рекомбинационных событий
- 1. 2. 3. 2. Альтернативный путь кроссоверной рекомбинации
- 112. 3. 3. ЯесА белгаги их роль в рекомбинации.'
- 1. 2. 3. 4. Выявление рекомбинации на цитологическом уровне
- 1. 3. Взаимосвязь ключевых событий профазы
- 1. 3. 1. Передислокация доменов хромосом и связь этого процесса с синапсисом и рекомбинацией
- 1. 3. 2. Связь процессов выравнивания, спаривания и синапсиса гомологичных хромосом с рекомбинацией
- 1. 3. 3. Синапсис, рекомбинация и интерференция
- 2. 1. Выделение мейотических мутантов из популяций ржи н их, поддержание в генетической коллекции
- 2. 2. Анализ наследования аномалий мейоза
- 23. Сравнительная цитологическая характеристика^ мейотических, мутантов у ржи
- 2. 3. 11. Методы цитологического анализа
- 2. 3. 2. Синаптические мутанты
- 2. 3. 3. Мутации, нарушающие конденсацию мейотических хромосом
- 2. 3. 4. Мутации, деформирующие латеральные элементы CK
- 2. 4. Картирование мейотических мутаций у ржи
- 214. 1. Анализ биохимических и молекулярных маркеров
- 2. 4. 2. Локализация синаптических мутаций, тест на аллелизм и выявление генотипов с разным сочетанием аллелей мей-генов.:. .95*
- 2. 5. Перевод.коллекции мейотических мутантов ржип на яровую основу
- 3. 1. Специфические особенности используемой модели
- 3. 1. 1. Характеристика растений, использованных для FISH
- 3. 1. 2. Определение и описание стадий мейоза
- 3. 1. 4. Зонды ДНК
- 3. 1. 5. Флуоресцентная ДНК-ДНК гибридизация in situ (FISH)
- 3. 1. 6. Идентификация ядер клеток в интервале стадий «предмейотическая интерфаза-лептотена»
- 3. 2. Типы агрегации субтеломерных иперицентромерных доменов хромосом у ржи дикого типа иудвухасинаптпческих мутантов! syl' и sy
- 3. 2. 1. Предмейотическая интерфаза — лептотена
- 3. 2. 2: Лептотена — зиготена
- 3. 2. 3. — Зиготена-пахитена^
- 3. 2. 4. Диплотена — диакинез- метафаза!
- 3. 3. Фенотип, обусловленный мутациями в генах синапсиса, может проявиться до образования синаптонемного комплекса
- Глава 4. СВЯЗЬ МЕЖДУ СИНАПСИСОМ И РЕКОМБИНАЦИЕЙ В
- 4. 1.1. Иммуноцитохимическая локализация белков Rad51 и Dmcl в микроспороцитах ржи
- 4. 1. *-2. Выявление мейотических узелков для хшмуноцитохимической локализации белков рекомбинации
- 411. 3. Клонирование и секвенирование генов рекомбинации^
- 4. 2. Локализация" рекомбиногенных белков Rad51 H^Dmcl в ядре и на хромосомах у синаптнческих мутантов
- 4. 3. Мейотические узелки
- 4. 3. 1. Ранние мейотические узелки
- 4. 3. 2. Связь между ранними, и поздними узелками
- 4. 3. 3. Поздние мейотические (рекомбинационые) узелки
- 4. 3.4. Мейотические узелки у ржи
- 4. 4. Анализ нуклеотидных последовательностей мей-генов ржи
- 4. 5. Сравнителтный анализ синапсиса и рекомбинации у растений и у почкующихся дрожжей S. cerevisiae (Обсуждение)
- 5. 1. Материал и методы
- 5. 2. Взаимодействие хромосом при^индискриминантном синапсисе
- 5. 3. Подходы к анализу рекомбинациши интерференции
- 6. 1. Материал и методы
- 6. 1. 1. Получение дисомных дополненных ржано-пшеничных линий, линий пшеницы, дополненных двумя телосомами 5RL ржи
- 6. 1. 2. Сбор пыльников и определение стадий мейоза в них
- 6. 1. 3. С-окраска
- 6. Г. 4. Геномная>гибридизация"ДНК-ДНК in situ (GISH)
- 6. 2. Эффект локуса Phi пшеницы на организацию хроматина и спаривание хромосом
- 6. 2. 1. Конфигурации, образуемые хромосомами на стадии метафазы
- 6. 2. 2. Влияние Phi на общую организацию хроматина
- 6. 2. 3. Морфологические особенности и поведение телосом 5RL
- 6. 2. 3. 1. Фенотип, соответствующий PhlPhl
- 6. 2. 3. 2. Фенотип, соответствующийphlbphlb
- 6. 2. Эффект локуса Phi пшеницы на организацию хроматина и спаривание хромосом
- 6. 3. Эффекты Phlb и взаимодействие хромосом (Обсуждение)
- 6. 3. 1. Влияние Phi на конденсацию хромосом
- 6. 3. 2. Влияние Phi на (пред)мейотическое спаривание хромосом
- 6. 3. 3. Влияние Phi на немейотическое спаривание хромосом
- 7. 1. Материал и методы
- 7. Г-1. Иммуноцитохимические эксперименты
- 7. 1. 2. Электронная микроскопия
- 7. 2. Протеомный «портрет» мутанта sylO
- 7. 2. 1. Изучение фенотипа sylO на ультраструктурном уровне
- 7. 2. 2. Белок синаптонемного комплекса^урГ и ассоциированный с CK белок Asyl образуют оси мейотических хромосом у sylO
- 7. 3. Двойные мутанты по синаптическнм генам SY1 и SY9 у ржи и их изучение методами молекулярной цитогенетики
- 7. 3. 1. Получение двойных мутантов sylsylsy9sy
- 7. 3. 2. Иммуноцитохимическая локализация белка Asyl в ядрах микроспороцитов асинаптических мутантов syl, sy9 и sylsy
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