Конструирование рекомбинантного штамма Escherichia coli, продуцирующего адгезин К88 и В-субъединицу холерного токсина
Во-первых, получают различные типы оральных цельноклеточных убитых вакцин, содержащих адгезивные антигены (Гутковский А. А., Дворкин Г. Л. 198?- Пирожков М. К., Тугаринов О. А. 1991; Соколова Н. А. ссоавт. 1993; Федоров Ю. Н. с соавт. 1997; Buttenshon Р.А. 1986; Gassier R.W. 1988). Теоретическим обоснованием для разработки таких вакцин служит тот факт, что развитие инфекционного процесса при… Читать ещё >
Содержание
- ВОЗБУДИТЕЛЯ К0ЛИБАКТЕРИ03 А
- 1. 1. 1. Характеристика колибактериозов
- 1. 1. 2. Основные этапы инфекционного процесса при коли-диареях
- 1. 1. 3. Факторы патогенности энтеротоксигенных штаммов кишечной палочки (ЕТЕС)
- 7. 1.1.4. Иммуногенный потенциал протективных антигенов энтеротоксигенных штаммов кишечной палочки
- 1. 2. Средства специфической профилактики диарейных заболеваний, вызываемых энтеротоксигенными штаммами кишечной палочки
- СОВСТВЕШШ ЙССЖД01АНЙЯ ГЛАВА ?. МАТЕРИАЛЫ И МЕТОДЫ
- 2. 1. Бактериальные штаммы и плазмиды
- 2. 2. Питательные среды и реактивы
- 2. 3. Методы
- 2. 3. 1. Коньюгационные скрещивания
- 2. 3. 2. Изучение стабильности наследования плазмид в клетках кишечной палочки методом популяцион-ного анализа
- 2. 3. 3. Изучение плазмидного состава полученных транс-коньюгантов Е. col
- 2. 3. 4. Реакция маннозорезистентной гемагглютинации
КОНСТРУИРОВАНИЕ ДШЛАЗМИДН0Г0 ШТАМПА КИШЕЧНОЙ ПАЛОЧКИ М1?(рК88:mini-kan)(pIEMS), СОДЕРЖАЩЕГО РЕКОМВИНАЫТНЫЕ ПЛАЗМИДЫ С КЛОНИРОВАННЫМИ ГЕНАМИ АДРЕЗИНА К88 И В-СУБЬЕДИНШЩ ХОЛЕРНОГО ТОКСИНА. 50. Коньюгациошюе введение в штамм кишечной палочки М17 плазмиды рК88: :rnlnl-kan и изучение стабильности ее наследования в клетках Е. coli М17. Изучение экспрессии плазмидных генов fas в клетках созданного штамма кишечной палочки М17(рК88:mini-kan)(pIEMS) и определение оптимальных условий для биосинтеза клетками адгезина К88.
3.3. Коньюгационное введение в штамм кишечной палочки М17(рК88: :iriini-kan) плазмиды р1ЕМЗ и изучение стабильности ее наследования в клетках Е. coli
М17(рК88:mini-kan) (PIEM3).
ГЛАВА 4. ИЗУЧЕНИЕ ЭКСПРЕССИИ И СТАБИЛЬНОСТИ НАСЛЕДОВАНИЯ ВВЕДЕННЫХ ГЕНОВ ПРОТЕКТИВНЫХ АНТИГЕНОВ — АДГ ЕВИНА К88 И В-СУБЬЕЯИНИЦЫ ХОЛЕРНОГО ТОКСИНА В ШТАММЕ E. coli М17(рК88:mini-kan).(рIEM3)
4.1. Определение экспрессии генов В-субьединицы холерного токсина в клетках штамма E. coli М17(рК88:mini-kan) (р1ЕМЗ).
4.2. Определение экспрессии генов адгезина К88 в клетках ?.coli М17(рК88:mini-kan)(pIEM3).
4.3. Стабильность наследования плазмид рК88: mini-kan и р1ЕМЗ штаммом E. coli М17 в условиях in vitro и in vivo.
ГЛАВА 5. ИЗУЧЕНИЕ БЕЗОПАСНОСТИ, КОЛОНИЗИРУЮЩЕЙ СПОСОБНОСТИ И АНТИГЕННЫХ СВОЙСТВ СКОНСТРУИРОВАННОГО ДИПЛАЗМИД-НОГО ШТАММА E. coli М17(рК88:mini-kan)(pIEM3). 86 5.1. Изучение безвредности штамма Е. coli М17(рК88:mini-kan) (pIEM3).
5.2. Определение колонизирующей способности штамма
Е. coli М17 (рК88:mini-kan) (pIEM3).
5.3. Изучение антигенных свойств штамма E. coli М17(рК88: mini-kan) (pIEM3).
ОБСУЖДЕНИЕ.
ВЫВОДЫ.
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