Идентификация генов ?-эндотоксинов и типирование штаммов B.thuringiensis методами генетического анализа
Диссертация
Выяснено, что токсичность в отношении насекомых отряда Чешуекрылые проявляли только штаммы с крупными и средними бипирамидальными (ромбовидными) кристаллами. Штаммы с мелкими кристаллами такой же формы были неактивны в отношении этих насекомых. Показано, что мелкие бипирамидальные кристаллы характерны для штаммов, содержащих гены сгу7, 8, которые кодируют токсины, активные против насекомых… Читать ещё >
Содержание
- СПИСОК УСЛОВНЫХ СОКРАЩЕНИЙ И ОБОЗНАЧЕНИЙ
- 1. ВВЕДЕНИЕ
- 2. ОБЗОР ЛИТЕРАТУРЫ
- 2. 1. Энтомопатогенная бактерия B. thuringiensis
- 2. 2. Классификация подвидов B. thuringiensis
- 2. 3. Токсины B. thuringiensis
- 2. 3. 1. Композиция кристаллов 5-эндотоксинов и их морфология
- 2. 3. 2. Доменная организация эндотоксинов
- 2. 3. 3. Механизм действия 5-эндотоксинов
- 2. 3. 4. Гены 5-эндотоксинов
- 2. 3. 5. Классификация 5-эндотоксинов
- 2. 4. Применение В. thuringiensis
- 2. 4. 1. Применение препаратов В. thuringiensis в качестве биопестицидов
- 2. 4. 2. Трансгенные растения, несущие гены энтомопатогенных белков В. thuringiensis
- 2. 4. 3. Медицинское применение штаммов В. thuringiensis
- 2. 5. Устойчивость популяций насекомых к действию B. thuriniensis
- 2. 5. 1. Лабораторные линии насекомых, устойчивые к действию В. thuringiensis
- 2. 5. 2. Природные линии насекомых, устойчивые к действию В. thuringiensis
- 2. 5. 3. Молекулярные механизмы устойчивости
- 2. 6. Скрининг коллекций для выявления перспективных штаммов В. thuringiensis
- 2. 6. 1. Выявление белков 5-эндотоксинов с помощью иммунологического анализа
- 2. 6. 2. Гибридизационный анализ геномной ДНК
- 2. 6. 3. ПЦР-анализ в определении генов энтомопатогенных белков.&bdquo-48 2.7. Методы классификации штаммов B. thuringiensis
- 2. 7. 1. Секвенирование последовательностей рРНК
- 2. 7. 2. Риботипирование рРНК (гибридизация)
- 2. 7. 3. Методы типирования, основанные на ПЦР
- 3. 1. Материалы
- 3. 1. 1. Реактивы
- 3. 1. 2. Ферменты
- 3. 1. 3. Питательные среды
- 3. 1. 4. Буферные растворы
- 3. 1. 5. Олигонуклеотиды
- 3. 1. 6. Бактерии
- 3. 1. 7. Насекомые
- 3. 2. Методы
- 3. 2. 1. Выделение ДНК
- 3. 2. 2. ПЦР-анализ по выявлению генов, кодирующих 5-эндотоксины
- 3. 2. 3. RAPD-анализ с праймерами GDI, GD
- 3. 2. 4. RAPD-анализ с праймером
- 3. 2. 5. Филогенетический анализ
- 3. 2. 6. Приготовление культур бактерий для биоанализа
- 3. 2. 7. Анализ белков методом электрофореза в SDS-ПААГ
- 3. 2. 8. Биотестирование штаммов
- 4. 1. Сравнительная характеристика штаммов с различной формой кристаллов 5-эндотоксинов
- 4. 1. 1. Штаммы с бипирамидальными крупными кристаллами
- 4. 1. 2. Штаммы с мелкими бипирамидальными кристаллами
- 4. 1. 3. Штаммы с бесформенными кристаллами
- 4. 1. 4. Акристаллические штаммы
- 4. 1. 5. Штаммы с нетипичной формой кристаллов
- 4. 2. Типирование штаммов В. thuringiensis методом RAPD-анализа
- 4. 2. 1. Подбор праймеров для RAPD-анализа
- 4. 2. 2. Анализ подвидов В. thuringiensis по RAPD-типам
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