Бесплатный автореферат и диссертация по биологии на тему

Конструирование и изучение рекомбинантных аденовирусов CELO в экспериментах in vitro и in vivo

ВАК РФ 03.00.03, Молекулярная биология

Заключение Диссертация по теме "Молекулярная биология", Шмаров, Максим Михайлович

ВЫВОДЫ.

1. Разработана методика получения рекомбинантных аденовирусов на основе генома аденовируса птиц CELO, основанная на гомологичной рекомбинации фрагментов вирусного генома в клетках LMH. Отработаны оптимальные условия получения препаративных количеств рекомбинантного аденовируса птиц CELO в куриных эмбрионах. Инфекционный титр препаратов вируса после концентрирования и очистки составил 1011 БОЕ/мл.

2. Получен набор рекомбинантных аденовирусов птиц CELO, содержащих репортерный ген GFP, а также гены функционального и дефектного рибозимов, направленных на расщепление мРНК гена РВ1 вируса гриппа А.

3. Впервые была показана трансдукция рекомбинантным аденовирусом птиц CELO-GFP культур клеток человека линий Н1299, НерЗВ и культур клеток животных линий MDBK, MDCK, COSI, RAT2, В16.

4. В экспериментах на лабораторных животных показано, что введение в хвостовую вену мыши рекомбинантного аденовируса CELO-GFP приводит к экспрессии гена GFP в клетках печени; впервые показана трансдукция клеток В16 меланомы у мышей при внутриопухолевом введении рекомбинантного вируса CELO-GFP.

5. Впервые показано ингибирование репродукции вируса гриппа А штамма A/WSN/33 в клетках линии А54 9, инфицированных рекомбинантным аденовирусом птиц CEL04D-N, экспрессирующим ген рибозима, направленного на расщепление мРНК гена РВ1 вируса гриппа А. Уровень подавления репродукции вируса гриппа составил 61,5%.

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Библиография Диссертация по биологии, кандидата биологических наук, Шмаров, Максим Михайлович, Москва

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