Модификация термостабильных ДНК-полимераз для повышения эффективности синтеза ДНК
Диссертация
В настоящей диссертационной работе представлении результаты исследования различий в свойствах Тая и ТШ ДНК-полимераз и их причин. Полученные данные позволили более рационально подайти к использоваию этих ферментов для решения прикладных задач и создали предпосылки для направленной корекции свойств этих полимераз. В результате была создана ДНК-полимераза способная высокоспецифично и эффективно… Читать ещё >
Содержание
- 1. Введение
- 2. Обзор литературы
- 2. 1. Принципы пространственной организации и механизма действия полимеразного домена ДНК-полимераз
- 2. 1. 1. Типы ДНК-полимераз
- 2. 1. 2. Принципы пространственной организации полимеразного домена
- 2. 1. 3. Механизм ферментативного синтеза ДНК. Субдомены «ладонь» и «пальцы»
- 2. 1. 4. Субдомен «большой палец»
- 2. 2. ДНК-полимеразы, используемые в генной инженерии и области их применения
- 2. 2. 1. ДНК-полимеразы из мезофильных организмов
- 2. 2. 1. 1. ДНК-полимераза IЕ. coli, Фрагмент Кленова и ДНК-полгшераза фага Т
- 2. 2. 1. 2. РНК-зависимые ДНК-полимеразы
- 2. 2. 1. 3. Секвенирование ДНК по методу Сенгера. ДНК-полимераза фага Т7 и ее модификации
- 2. 2. 2. Полимеразная цепная реакция (ПЦР)
- 2. 2. 3. Термостабильные ДНК-полимеразы
- 2. 2. 3. 1. ДНК-полимераза I Thermus aquaticus (Тaq-полимераза) и ее модификации
- 2. 2. 3. 2. ДНК-полимераза I Thermus thermophilus (Tth-полимераза)
- 2. 2. 3. 3. Термостабильные ДНК-полимеразы с 3'-5' экзонуклеазной активностью (Vent- и Pfu- полимеразы)
- 2. 2. 3. 4. Ферментативные смеси для ПЦР-амплификации длинных последовательностей ДНК
- 2. 2. 1. ДНК-полимеразы из мезофильных организмов
- 2. 1. Принципы пространственной организации и механизма действия полимеразного домена ДНК-полимераз
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