Автор работы: Пользователь скрыл имя, 27 Июня 2012 в 14:02, дипломная работа
Работа посвящена вопросу о флуоресцентных белках и генной инженерии.В работе кратко изложена история открытия белков их биологическое и генетическое значение в современном мире. Так же рассказывается о работе по молекулярной биологии с использованием флуоресцентных белков. Работа защищена на отлично.
ВВЕДЕНИЕ 4-7
ОБЗОР ЛИТЕРАТУРЫ 8-51
1. Стволовые клетки 8-36
1.1. Понятие о стволовых клетках 8-10
1.2. Классификация стволовых клеток 10-13
1.3. Свойства стволовых клеток 13-17
1.4. Особенности региональных нейральных стволовых клеток 17-20
1.5. Стволовые клетки, как удобная модель для анализа роли генов в процессе дифференцировки 20-21
1.6. Стволовые клетки перспективы использования в медицине 22-25
1.7.Генная терапия 26-36
1.7.1. Понятие о генной терапии 26-27
1.7.2. Условия проведения генной терапии 27-30
1.7.3.Проблема генной и клеточной терапии 31-33
1.7.4. Лечение заболеваний с помощью генной терапии 33-36
2.Флуоресцентные белки 36-47
2.1. Открытие зеленого флуоресцентного белка 36
2.2. Биохимические, спектральные и физические свойства GFP 36-37
2.3. Трехмерная структура GFP 37-41
2.4. Трехмерная структура DsRed 41-44
2.5. Использование GFP-подобных белков для изучения подвижности клеток, клеточных белков и органелл 44-47
3.Векторы 47-48
3.1. Вектор pcDNA 3.1+ 48-49
3.2. CMV – промотор 49-50
4. Заключение 51
МАТЕРИАЛЫ И МЕТОДЫ 52-55
Ферменты и реактивы 52
Приготовление компетентных клеток 52
Трансформация бактерий E.coli XL I-Blue плазмидами 52-53
Выделение плазмидной ДНК из E.coli 53
Электрофорез в агарозном геле 53
Рестрикция 53
Контроль наличия вставки (блот-гибридизация по Саузерну). 54-55
Достраивание липких концов ДНК до тупых 55
Лигирование 55
Конструкции 55
РЕЗУЛЬТАТЫ 56-66
Обсуждение результатов 67-68
Выводы 69
Список сокращений
Список литературы 70-79
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