RU2012115127A - Трансгенный объект coи mon 87708 и способы его применения - Google Patents

Трансгенный объект coи mon 87708 и способы его применения Download PDF

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RU2012115127A
RU2012115127A RU2012115127/10A RU2012115127A RU2012115127A RU 2012115127 A RU2012115127 A RU 2012115127A RU 2012115127/10 A RU2012115127/10 A RU 2012115127/10A RU 2012115127 A RU2012115127 A RU 2012115127A RU 2012115127 A RU2012115127 A RU 2012115127A
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soybean
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Рональд Дж. БРИНКЕР
Уэн К. БЕРНС
Пол С.С. Фэн
Анджу ГУПТА
Шио-Вай Хой
Марианне МАЛВЕН
Куншенг ВУ
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МОНСАНТО ТЕКНОЛОДЖИ ЭлЭлСи
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Abstract

1. Рекомбинантная молекула ДНК, содержащая нуклеотидную молекулу, содержащую нуклеотидную последовательность, выбранную из группы, состоящей из SEQ ID NO: 1-8, и комплементарных им последовательностей.2. Рекомбинантная молекула ДНК по п.1, где указанная рекомбинантная молекула ДНК образована инсерцией гетерологичной молекулы нуклеиновой кислоты в геномную ДНК растения сои, растительной клетки или семени.3. Рекомбинантная молекула ДНК по п.1, где указанную молекулу ДНК получают из трансгенного объекта сои MON 87708, показательный образец которого депонирован в ATCC как PTA-9670.4. Рекомбинантная молекула ДНК по п.1, где указанная молекула ДНК находится в растении сои, растительной клетке, семени, части растения или товаре, полученным из объекта сои MON 87708, показательный образец которого депонирован в ATCC как PTA-9670.5. Рекомбинантная молекула ДНК по п.1, где указанная молекула ДНК представляет собой ампликон, диагностический в отношении наличия полученной из объекта MON 87708 ДНК.6. Молекула ДНК, содержащая нуклеотидную молекулу, обладающую нуклеотидной последовательностью с достаточной длиной непрерывной нуклеотидной последовательности SEQ ID NO: 6 и комплементарной ей последовательности, для функционирования в качестве ДНК-зонда, гибридизующегося в строгих условиях гибридизации с молекулой ДНК, содержащей нуклеотидную последовательность, выбранную из группы, состоящей из SEQ ID NO: 1-8, и негибридизующегося в строгих условиях гибридизации с молекулой ДНК, не содержащей нуклеотидную последовательность, выбранную из группы, состоящей из SEQ ID NO: 1-8.7. Пара молекул ДНК, состоящая из первой молекулы ДНК и второй молекулы ДНК, отличающейся от перво

Claims (30)

1. Рекомбинантная молекула ДНК, содержащая нуклеотидную молекулу, содержащую нуклеотидную последовательность, выбранную из группы, состоящей из SEQ ID NO: 1-8, и комплементарных им последовательностей.
2. Рекомбинантная молекула ДНК по п.1, где указанная рекомбинантная молекула ДНК образована инсерцией гетерологичной молекулы нуклеиновой кислоты в геномную ДНК растения сои, растительной клетки или семени.
3. Рекомбинантная молекула ДНК по п.1, где указанную молекулу ДНК получают из трансгенного объекта сои MON 87708, показательный образец которого депонирован в ATCC как PTA-9670.
4. Рекомбинантная молекула ДНК по п.1, где указанная молекула ДНК находится в растении сои, растительной клетке, семени, части растения или товаре, полученным из объекта сои MON 87708, показательный образец которого депонирован в ATCC как PTA-9670.
5. Рекомбинантная молекула ДНК по п.1, где указанная молекула ДНК представляет собой ампликон, диагностический в отношении наличия полученной из объекта MON 87708 ДНК.
6. Молекула ДНК, содержащая нуклеотидную молекулу, обладающую нуклеотидной последовательностью с достаточной длиной непрерывной нуклеотидной последовательности SEQ ID NO: 6 и комплементарной ей последовательности, для функционирования в качестве ДНК-зонда, гибридизующегося в строгих условиях гибридизации с молекулой ДНК, содержащей нуклеотидную последовательность, выбранную из группы, состоящей из SEQ ID NO: 1-8, и негибридизующегося в строгих условиях гибридизации с молекулой ДНК, не содержащей нуклеотидную последовательность, выбранную из группы, состоящей из SEQ ID NO: 1-8.
7. Пара молекул ДНК, состоящая из первой молекулы ДНК и второй молекулы ДНК, отличающейся от первой молекулы ДНК, где каждая из указанных первой и второй молекул ДНК содержит нуклеотидную молекулу, обладающую нуклеотидной последовательностью с достаточной длиной непрерывных нуклеотидов SEQ ID NO: 6 или комплементарной ей последовательности, для функционирования в качестве ДНК-праймеров при применении в реакции амплификации с ДНК, полученной из объекта MON 87708, для получения в образце ампликона, характерного для ДНК объекта сои MON 87708.
8. Способ определения в образце наличия молекулы ДНК, полученной из объекта сои MON 87708, где указанный способ включает:
a. приведение образца в контакт с ДНК-зондом по п.6;
b. подвергание указанного образца и указанного ДНК-зонда строгим условиям гибридизации; и
c. детектирование гибридизации указанного ДНК-зонда с молекулой ДНК в указанном образце, где гибридизация указанного ДНК-зонда с указанной молекулой ДНК свидетельствует о наличии в указанном образце молекулы ДНК, полученной из объекта сои MON 87708.
9. Способ определения в образце наличия молекулы ДНК, полученной из объекта сои MON 87708, где указанный способ включает:
a. приведение образца в контакт с парой молекул ДНК по п.7;
b. осуществление реакции амплификации, достаточной для получения ДНК-ампликона, содержащего последовательность, выбранную из группы, состоящей из SEQ ID NO: 1-8 и комплементарные им последовательности; и
c. детектирование наличия указанного ДНК-ампликона в указанной реакции, где присутствие указанного ДНК-ампликона в указанной реакции свидетельствует о наличии в указанном образце молекулы ДНК, полученной из объекта сои MON 87708.
10. Набор для детекции ДНК, содержащий, по меньшей мере, одну молекулу ДНК, содержащую нуклеотидную последовательность с достаточной длиной непрерывной нуклеотидной последовательности SEQ ID NO: 6 и комплементарную ей последовательность, для функционирования в качестве ДНК-праймера или зонда, специфичного для определения наличия ДНК, полученной из объекта сои MON 87708, где детекция указанной ДНК является диагностической в отношении наличия в образце указанной ДНК объекта MON 87708.
11. Рекомбинантное растение сои, семя, клетка или часть этого растения, содержащие нуклеотидную молекулу, обладающую нуклеотидной последовательностью, выбранной из группы, состоящей из SEQ ID NO: 1-8 и комплементарных им последовательностей.
12. Рекомбинантное растение сои, семя, клетка или часть этого растения по п.11, где указанное растение, семя, клетка или часть этого растения являются устойчивыми к обработке гербицидом дикамба.
13. Рекомбинантное растение сои, семя, клетка или часть этого растения по п.11, с помощью генома которого получают ампликон, содержащий молекулу ДНК, выбранную из группы, состоящей из SEQ ID NO: 1-8 и комплементарных им последовательностей, при тестировании в способе амплификации ДНК.
14. Растение сои или семя, где указанное растение сои или семя получают из объекта сои MON 87708, показательный образец которого депонирован в ATCC с номером доступа № PTA-9670.
15. Растение сои или семя по п.14, где указанное растение сои или семя представляет собой гибрид, обладающий, по меньшей мере, одним родителем, полученным из объекта сои MON 87708.
16. Неживой растительный материал, содержащий рекомбинантную молекулу ДНК, выбранную из группы, состоящей из SEQ ID NO: 1-8 и комплементарных им последовательностей.
17. Микроорганизм, содержащий нуклеотидную молекулу, обладающую нуклеотидной последовательностью, выбранной из группы, состоящей из SEQ ID NO: 1-8 и комплементарных им последовательностей.
18. Микроорганизм по п.17, где указанный микроорганизм представляет собой растительную клетку.
19. Товар, содержащий нуклеотидную молекулу, обладающую нуклеотидной последовательностью, выбранной из группы, состоящей из SEQ ID NO: 1-8 и комплементарных им последовательностей.
20. Товар по п.19, где указанный товар выбран из группы, состоящей из целых или обработанных семян, корма для животных, масла, муки грубого помола, муки, хлопьев, отрубей, молока, сыра, бумаги, сливок, вина, биомассы и топливных продуктов.
21. Способ контроля сорняков в поле, включающий посев растений объекта сои MON 87708 в поле и нанесение эффективной дозы гербицида дикамба для контроля сорняков на указанном поле без повреждения указанных растений объекта сои MON 87708.
22. Способ по п.21, где указанная эффективная доза гербицида дикамба составляет от приблизительно 0,005 фунтов до приблизительно 8 фунтов на акр.
23. Способ контроля сорняков в поле, включающий нанесение эффективной дозы гербицида дикамба для контроля сорняков в поле и затем посев объекта сои MON 87708 в указанное поле.
24. Способ по п.23, где указанная эффективная доза гербицида дикамба составляет от приблизительно 0,005 фунтов до приблизительно 8 фунтов на акр, и указанный посев объекта сои MON 87708 осуществляют в течение 14 дней от указанного нанесения эффективной дозы гербицида дикамба.
25. Способ получения семян сои, по существу не содержащих семян вредоносных видов сорняков, где указанный способ включает:
a. посев семян объекта сои MON 87708 в поле;
b. нанесение эффективной дозы гербицида дикамба на указанное поле для уничтожения вредоносных сорняков на указанном поле без повреждения указанных растений объекта сои MON 87708; и
c. сбор семян сои с указанного поля.
26. Способ по п.25, где указанные семена вредоносных видов сорняков выбраны из группы, состоящей из Cardaria spp., Heliotropium spp., Centaurea spp., Senecio spp., Crotalaria spp., Solanum spp., Xanthium spp., Amsinckia spp., Cassia spp., Sesbania spp., Datura spp., Ricinus spp., Argemone spp., Corchorus spp., Impomoea spp. и Echium spp.
27. Способ получения растения сои, устойчивого к нанесению гербицида дикамба, включающий:
a. половое скрещивание трансгенного растения объекта сои MON 87708, содержащего нуклеотидную молекулу, обладающую нуклеотидной последовательностью, выбранной из группы, состоящей из SEQ ID NO: 1-8 и комплементарных им последовательностей, со вторым растением сои;
b. сбор семян, полученных от указанного скрещивания;
c. выращивание указанных семян для получения множества растений-потомков;
d. обработку указанного множества растений-потомков дикамбой; и
e. селекцию растения-потомка, устойчивого к дикамбе.
28. Способ получения растения сои, устойчивого к нанесению гербицида дикамба, включающий:
a. самоопыление трансгенного растения объекта сои MON 87708, содержащего нуклеотидную молекулу, обладающую нуклеотидной последовательностью, выбранной из группы, состоящей из SEQ ID NO: 1-8 и комплементарных им последовательностей;
b. сбор семян, полученных от указанного самоопыления;
c. выращивание указанных семян для получения множества растений-потомков;
d. обработку указанного множества растений-потомков дикамбой; и
e. селекцию растения-потомка, устойчивого к дикамбе.
29. Способ определения зиготности растения или семян объекта сои MON 87708, включающий:
a. приведение образца, содержащего ДНК сои, в контакт с набором праймеров, содержащим SEQ ID NO: 12, SEQ ID NO: 13 и SEQ ID NO: 14, и набором зондов, содержащим SEQ ID NO: 15 и SEQ ID NO: 16;
b. осуществление реакции амплификации нуклеиновой кислоты с указанным образцом, набором праймеров и набором зондов;
c. детектирование в указанной реакции амплификации нуклеиновой кислоты первого флуоресцентного сигнала, характерного для объекта MON 87708, и второго флуоресцентного сигнала, отличающегося от указанного первого флуоресцентного сигнала и диагностического в отношении геномной ДНК природной сои, соответствующей положению инсерции трансгена объекта MON 87708; и
d. анализ наличия и/или отсутствия указанного первого флуоресцентного сигнала и указанного второго флуоресцентного сигнала в указанной реакции амплификации нуклеиновой кислоты, где присутствие обоих флуоресцентных сигналов свидетельствует о том, что указанный образец является гетерозиготным по объекту MON 87708, и наличие только указанного первого флуоресцентного сигнала свидетельствует о том, что указанный образец является гомозиготным по объекту MON 87708.
30. Область гаплотипа сои в группе сцепления 9 приблизительно в положении 143.5, содержащая ген устойчивости к дикамбе и дополнительно определяемая по "скользящему окну" гаплотипа 19743 и 19767.
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