RU2009143751A - Производство белков с высоким содержанием маннозы в культуре растительных клеток - Google Patents

Производство белков с высоким содержанием маннозы в культуре растительных клеток Download PDF

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RU2009143751A
RU2009143751A RU2009143751/10A RU2009143751A RU2009143751A RU 2009143751 A RU2009143751 A RU 2009143751A RU 2009143751/10 A RU2009143751/10 A RU 2009143751/10A RU 2009143751 A RU2009143751 A RU 2009143751A RU 2009143751 A RU2009143751 A RU 2009143751A
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nucleic acid
human
galactosidase
seq
protein
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Йозеф ШАЛТИЛ (IL)
Йозеф ШАЛТИЛ
Гидеон БАУМ (IL)
Гидеон Баум
Даниел БАРТФЕЛД (IL)
Даниел БАРТФЕЛД
Шарон ХАШМУЕЛИ (IL)
Шарон Хашмуели
Айала ЛЕВКОВИЧ (IL)
Айала ЛЕВКОВИЧ
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Проталикс Лтд (Il)
Проталикс Лтд
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Abstract

1. Выделенная нуклеиновокислотная последовательность, кодирующая человеческий лизосомальный белок, непосредственно соединенный с С-концевым сигналом вакуолярного нацеливания и N-концевым сигнальным пептидом эндоплазматического ретикулума, отличающаяся тем, что данный человеческий лизосомальный белок является человеческой α-галактозидазой. ! 2. Выделенная нуклеиновокислотная последовательность, кодирующая человеческий лизосомальный белок, непосредственно соединенный с С-концевым сигналом удерживания в эндоплазматическом ретикулуме и N-концевым сигнальным пептидом эндоплазматического ретикулума, отличающаяся тем, что данный человеческий лизосомальный белок является человеческой α-галактозидазой. ! 3. Выделенная нуклеиновая кислота по пп.1 и 2, отличающаяся тем, что упомянутая человеческая α-галактозидаза имеет последовательность, обозначенную как SEQ ID NO:24. ! 4. Выделенный нуклеиновокислотный конструкт по п.1, отличающийся тем, что упомянутый сигнал вакуолярного нацеливания имеет последовательность, обозначенную как SEQ ID NO:4. ! 5. Выделенный нуклеиновокислотный конструкт по п.2, отличающийся тем, что упомянутый сигнал удерживания в эндоплазматическом ретикулуме имеет последовательность, обозначенную как SEQ ID NO:23 (KDEL). ! 6. Выделенный нуклеиновокислотный конструкт по п.2, имеющий последовательность, обозначенную как SEQ ID NO:19. ! 7. Выделенная нуклеиновая кислота по п.1, имеющая последовательность, обозначенную как SEQ ID NO:17. ! 8. Нуклеиновокислотный конструкт, способный к экспрессии в растительной клетке, содержащей выделенную нуклеиновую кислоту по п.1 или 2. ! 9. Клетка, содержащая нуклеиновокислотный конструкт

Claims (15)

1. Выделенная нуклеиновокислотная последовательность, кодирующая человеческий лизосомальный белок, непосредственно соединенный с С-концевым сигналом вакуолярного нацеливания и N-концевым сигнальным пептидом эндоплазматического ретикулума, отличающаяся тем, что данный человеческий лизосомальный белок является человеческой α-галактозидазой.
2. Выделенная нуклеиновокислотная последовательность, кодирующая человеческий лизосомальный белок, непосредственно соединенный с С-концевым сигналом удерживания в эндоплазматическом ретикулуме и N-концевым сигнальным пептидом эндоплазматического ретикулума, отличающаяся тем, что данный человеческий лизосомальный белок является человеческой α-галактозидазой.
3. Выделенная нуклеиновая кислота по пп.1 и 2, отличающаяся тем, что упомянутая человеческая α-галактозидаза имеет последовательность, обозначенную как SEQ ID NO:24.
4. Выделенный нуклеиновокислотный конструкт по п.1, отличающийся тем, что упомянутый сигнал вакуолярного нацеливания имеет последовательность, обозначенную как SEQ ID NO:4.
5. Выделенный нуклеиновокислотный конструкт по п.2, отличающийся тем, что упомянутый сигнал удерживания в эндоплазматическом ретикулуме имеет последовательность, обозначенную как SEQ ID NO:23 (KDEL).
6. Выделенный нуклеиновокислотный конструкт по п.2, имеющий последовательность, обозначенную как SEQ ID NO:19.
7. Выделенная нуклеиновая кислота по п.1, имеющая последовательность, обозначенную как SEQ ID NO:17.
8. Нуклеиновокислотный конструкт, способный к экспрессии в растительной клетке, содержащей выделенную нуклеиновую кислоту по п.1 или 2.
9. Клетка, содержащая нуклеиновокислотный конструкт по п.8.
10. Клетка по п.9, являющаяся растительной клеткой, отличающейся тем, что упомянутый белок - человеческая α-галактозидаза рекомбинантно продуцируется таким образом, что в его составе присутствуют, как минимум, один остаток ксилозы и, как минимум, один обнаженный маннозный остаток.
11. Клетка по п.9, являющаяся растительной клеткой, отличающейся тем, что упомянутый белок - человеческая α-галактозидаза рекомбинантно продуцируется таким образом, что в его составе присутствуют, как минимум, один остаток α-(1,2) ксилозы в гликановом остове и, как минимум, один остаток α-(1,3) фукозы в гликановом остове.
12. Выделенный белок - человеческая α-галактозидаза, кодируемый нуклеиновой кислотой по любому из пп.1-7, причем упомянутая человеческая α-галактозидаза захватывается фибровластами и обладает ферментативной активностью α-галактозидазы.
13. Белок - человеческая α-галактозидаза, продуцируемый клеткой по любому из пп.9-11.
14. Белок - человеческая α-галактозидаза, содержащий по крайней мере один элемент из группы, состоящей из:
(а) С-концевого сигнала удерживания в эндоплазматическом ретикулуме;
(б) N-концевого сигнального пептида эндоплазматического ретикулума;
(в) С-концевого сигнала вакуолярного нацеливания;
(г) по крайней мере одного остатка ксилозы и по крайней мере одного обнаженного маннозного остатка; и
(д) по крайней мере одного остатка α-(1,2) ксилозы в гликановом остове и по крайней мере одного остатка α-(1,3) фукозы в гликановом остове.
15. Использование белка - человеческой α-галактозидазы по любому из пп.12-14 с целью производства лекарственного препарата для лечения болезни Фабри.
RU2009143751/10A 2007-04-30 2008-04-30 Производство белков с высоким содержанием маннозы в культуре растительных клеток RU2009143751A (ru)

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Application Number Priority Date Filing Date Title
US11/790,991 2007-04-30
US11/790,991 US7951557B2 (en) 2003-04-27 2007-04-30 Human lysosomal proteins from plant cell culture

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