Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation
Lox sites of the Cre/lox recombination system from bacteriophage P1 were analyzed for their ability to affect on transgene expression when inserted upstream from a gene coding sequence adjacent to the right border (RB) of T-DNA. Wild and mutated types of lox sites were tested for their effect upon b...
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| Дата: | 2013 |
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| Автори: | , , , , |
| Формат: | Стаття |
| Мова: | English |
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Інститут клітинної біології та генетичної інженерії НАН України
2013
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| Назва видання: | Цитология и генетика |
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| Онлайн доступ: | http://dspace.nbuv.gov.ua/handle/123456789/126571 |
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| Назва журналу: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Цитувати: | Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation / N. Shcherbak, O. Kishchenko, L. Sakhno, I. Komarnytsky, M. Kuchuk // Цитология и генетика. — 2013. — Т. 47, № 3. — С. 21-32. — Бібліогр.: 48 назв. — англ. |
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irk-123456789-1265712017-11-27T03:02:49Z Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation Shcherbak, N. Kishchenko, O. Sakhno, L. Komarnytsky, I. Kuchuk, M. Оригинальные работы Lox sites of the Cre/lox recombination system from bacteriophage P1 were analyzed for their ability to affect on transgene expression when inserted upstream from a gene coding sequence adjacent to the right border (RB) of T-DNA. Wild and mutated types of lox sites were tested for their effect upon bar gene expression in plants obtained via Agrobacterium-mediated and biolistic transformation methods. Lox-mediated expression of bar gene, recognized by resistance of transgenic plants to PPT, occurred only in plants obtained via Agrobacterium-mediated transformation. RT-PCR analysis confirms that PPT-resistant phenotype of transgenic plants obtained via Agrobacterium-mediated transformation was caused by activation of bar gene. The plasmid with promoterless gus gene together with the lox site adjacent to the RB was constructed and transferred to Nicotiana tabacum as well. Transgenic plants exhibited GUS activity and expression of gus gene was detected in plant leaves. Expression of bar gene from the vectors containing lox site near RB allowed recovery of numerous PPT-resistant transformants of such important crops as Beta vulgaris, Brassica napus, Lactuca sativa and Solanum tuberosum. Our results demonstrate that the lox site sequence adjacent to the RB can be used to control bar gene expression in transgenic plants. Проанализирована способность lox-сайтов Cre/lox системы рекомбинации бактериофага Р1 влиять на экспрессию трансгенов при расположении этой последовательности непосредственно возле правого бордера (RB) перед кодирующей последовательностью гена. Нативная и мутированная последовательность lox-сайта были размещены в векторах для трансформации возле гена bar и проведена генетическая трансформация растений с помощью агробактерии и биолистическим методом. Lox-опосредованная экспрессия гена bar, обусловливающая устойчивость растений к фосфинотрицину, наблюдалась только у растений, которые получены с помощью агробактериальной трансформации. Методом РТ-ПЦР анализа подтверждено, что в трансгенных растениях, устойчивых к фосфинотрицину, происходит транскрипция гена bar. Сконструирован вектор, в котором ген gus и предшествующий ему lox-сайт размещены вблизи правого бордера, и проведена трансформация табака этим вектором. Экспрессия гена gus задетектирована в листьях трансгенных растений. Векторы, у которых последовательность lox-сайта предшествует гену bar возле правого бордера (RB-lox-bar), успешно использованы для получения устойчивых к фосфинотрицину трансгенных растений таких видов, как Beta vulgaris, Brassica napus, Lactuca sativa и Solanum tuberosum. Наши результаты подтверждают возможность использования последовательности lox-сайта возле правого бордера для контроля экспрессии гена bar в трансгенных растениях. 2013 Article Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation / N. Shcherbak, O. Kishchenko, L. Sakhno, I. Komarnytsky, M. Kuchuk // Цитология и генетика. — 2013. — Т. 47, № 3. — С. 21-32. — Бібліогр.: 48 назв. — англ. 0564-3783 DOI: 10.3103/S0095452713030079 http://dspace.nbuv.gov.ua/handle/123456789/126571 577.21:582.926.2 en Цитология и генетика Інститут клітинної біології та генетичної інженерії НАН України |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| language |
English |
| topic |
Оригинальные работы Оригинальные работы |
| spellingShingle |
Оригинальные работы Оригинальные работы Shcherbak, N. Kishchenko, O. Sakhno, L. Komarnytsky, I. Kuchuk, M. Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation Цитология и генетика |
| description |
Lox sites of the Cre/lox recombination system from bacteriophage P1 were analyzed for their ability to affect on transgene expression when inserted upstream from a gene coding sequence adjacent to the right border (RB) of T-DNA. Wild and mutated types of lox sites were tested for their effect upon bar gene expression in plants obtained via Agrobacterium-mediated and biolistic transformation methods. Lox-mediated expression of bar gene, recognized by resistance of transgenic plants to PPT, occurred only in plants obtained via Agrobacterium-mediated transformation. RT-PCR analysis confirms that PPT-resistant phenotype of transgenic plants obtained via Agrobacterium-mediated transformation was caused by activation of bar gene. The plasmid with promoterless gus gene together with the lox site adjacent to the RB was constructed and transferred to Nicotiana tabacum as well. Transgenic plants exhibited GUS activity and expression of gus gene was detected in plant leaves. Expression of bar gene from the vectors containing lox site near RB allowed recovery of numerous PPT-resistant transformants of such important crops as Beta vulgaris, Brassica napus, Lactuca sativa and Solanum tuberosum. Our results demonstrate that the lox site sequence adjacent to the RB can be used to control bar gene expression in transgenic plants. |
| format |
Article |
| author |
Shcherbak, N. Kishchenko, O. Sakhno, L. Komarnytsky, I. Kuchuk, M. |
| author_facet |
Shcherbak, N. Kishchenko, O. Sakhno, L. Komarnytsky, I. Kuchuk, M. |
| author_sort |
Shcherbak, N. |
| title |
Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation |
| title_short |
Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation |
| title_full |
Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation |
| title_fullStr |
Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation |
| title_full_unstemmed |
Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation |
| title_sort |
lox-dependent gene expression in transgenic plants obtained via agrobacterium-mediated transformation |
| publisher |
Інститут клітинної біології та генетичної інженерії НАН України |
| publishDate |
2013 |
| topic_facet |
Оригинальные работы |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/126571 |
| citation_txt |
Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation / N. Shcherbak, O. Kishchenko, L. Sakhno, I. Komarnytsky, M. Kuchuk // Цитология и генетика. — 2013. — Т. 47, № 3. — С. 21-32. — Бібліогр.: 48 назв. — англ. |
| series |
Цитология и генетика |
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| first_indexed |
2023-10-18T20:51:07Z |
| last_indexed |
2023-10-18T20:51:07Z |
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