RU2224024C2 - Трансгенные растения - Google Patents

Трансгенные растения Download PDF

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RU2224024C2
RU2224024C2 RU99116249/13A RU99116249A RU2224024C2 RU 2224024 C2 RU2224024 C2 RU 2224024C2 RU 99116249/13 A RU99116249/13 A RU 99116249/13A RU 99116249 A RU99116249 A RU 99116249A RU 2224024 C2 RU2224024 C2 RU 2224024C2
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seq
dna
plant
nucleotide sequence
amplification
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Марие МАННЕРЛЁФ (SE)
Марие МАННЕРЛЁФ
Пауль Петер ТЕННИНГ (SE)
Пауль Петер ТЕННИНГ
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Пер СТЕЕН
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Зингента Партисипейшнс Аг
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Abstract

Изобретение относится к генетической инженерии и может быть использовано в селекции растений. Опосредованная Agrobacterium трансформация растения сахарной свеклы, в котором экспрессируется фермент с ср4/epsps-активностью, придает этим растениям устойчивость к обработке гербицидом. 2 с. и 11 з.п. ф-лы, 7 табл.

Description

Текст описания в факсимильном виде (см. графическую часть)

Claims (13)

1. Способ получения трансгенного растения сахарной свеклы, включая его семена, у которого ПЦР-амплификация с использованием в качестве матрицы его геномной ДНК приводит к амплификации фрагмента ДНК размером 739 пар оснований при использовании пары олигонуклеотидных праймеров с последовательностями, представленными в SEQ ID NO: 18 и SEQ ID NO: 21, заключающийся в том, что осуществляют (а) трансформацию выращенных in vitro семядолей сахарной свеклы с помощью Agrobacterium, содержащих вектор, несущий участок ДНК, кодирующий ср4/epsps, (б) регенерацию ростков в присутствии глифосата, (в) перенос ростков в почву в теплице, (г) обработку проростков глифосатом, (д) визуальную оценку мощности растения и хлороза листьев по балльной шкале от 0 до 9, (е) отбор растений, у которых мощность растения и хлороз листьев оценена в 9 баллов, (ж) размножение отобранных растений с помощью обычных методов выведения сорта.
2. Способ по п.1, в котором ДНК, имеющая нуклеотидную последовательность, представленную в SEQ ID NO: 27, образует часть генома растения.
3. Способ по п.1, отличающийся тем, что ПЦР-амплификация с использованием в качестве матрицы его геномной ДНК приводит к амплификации фрагмента ДНК размером 834 пары оснований при использовании пары олигонуклеотидных праймеров с последовательностями, представленными в SEQ ID NO: 20 и SEQ ID NO: 24.
4. Способ по п.1, отличающийся тем, что ПЦР-амплификация с использованием в качестве матрицы его геномной ДНК, приводит к амплификации фрагмента ДНК размером 1057 пар оснований при использовании пары олигонуклеотидных праймеров с последовательностями, представленными в SEQ ID NO: 17 и SEQ ID NO: 22.
5. Способ по п.1, отличающийся тем, что ПЦР-амплификация с использованием в качестве матрицы его геномной ДНК, приводит к амплификации фрагмента ДНК размером 1224 пары оснований при использовании пары олигонуклеотидных праймеров с последовательностями, представленными в SEQ ID NO: 19 и SEQ ID NO: 25.
6. Способ по п.1, в котором ДНК, имеющая нуклеотидную последовательность, представленную в SEQ ID NO: 1, образует часть генома растения.
7. Способ по п.6, где нуклеотидная последовательность замещает последовательности ДНК с большим количеством повторов.
8. Способ по п.6, где части генома, непосредственно связанные с указанной нуклеотидной последовательностью, имеют нуклеотидные последовательности, представленные в SEQ ID NO: 2 и SEQ ID NO: 3.
9. Способ по п.7, в котором ДНК, имеющая нуклеотидную последовательность, представленную в SEQ ID NO: 4, образует часть генома растения.
10. Способ по п.1, предназначенный для получения семян, задепонированных под регистрационным номером NCIMB 97087006/2.
11. Способ по п.1, где вектор, кодирующий протеин ср4/epsps, представляет собой вектор, содержащий участок ДНК, который имеет нуклеотидную последовательность, представленную в SEQ ID NO: 5.
12. Способ по п.1, где регенерированные ростки анализируют с помощью ПЦР на наличие ДНК, кодирующей ср4/epsps.
13. Способ получения толерантного к глифосату растения сахарной свеклы, включая его потомство, включающий опосредованную трансформацию Agrobacterium с помощью ДНК, обеспечивающей экспрессию ср4/epsps, сообщающего устойчивость к глифосату растениям, при этом геномная ДНК растения лишена как правой, так и левой пограничных последовательностей Т-ДНК в области сайта интеграции.
RU99116249/13A 1997-10-31 1998-10-29 Трансгенные растения RU2224024C2 (ru)

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GB2337263A (en) 1999-11-17
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US6204436B1 (en) 2001-03-20
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HK1025360A1 (en) 2000-11-10
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CZ235199A3 (cs) 1999-09-15
DK199900916A (da) 1999-06-28
SE9902189L (sv) 1999-08-19
SE520353C2 (sv) 2003-07-01
HUP0001729A2 (hu) 2000-09-28
CN1242803A (zh) 2000-01-26
SE9902189D0 (sv) 1999-06-10
EP0950109A1 (en) 1999-10-20
DE19881833B4 (de) 2007-05-10
CA2279258A1 (en) 1999-05-14
AU1558099A (en) 1999-05-24
CN1148453C (zh) 2004-05-05
FI991221A (fi) 1999-05-28
HUP0001729A3 (en) 2002-04-29
CZ300208B6 (cs) 2009-03-18
US6531649B1 (en) 2003-03-11
PL334135A1 (en) 2000-02-14
FI991221A0 (fi) 1999-05-28

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