IP Library Granted Patent US 9,752,155
Granted Patent B2
US 9,752,155 · App. 14/472,977 · Granted Sep 5, 2017

Constructs for expressing transgenes using regulatory elements from brachypodium ubiquitin 1C genes

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Quick Facts
Patent No.
US 9,752,155
App. No.
14/472,977
Granted
Sep 5, 2017
Kind
B2
Abstract

Provided are constructs and methods for expressing a transgene in plant cells and/or plant tissues using the regulatory elements, including the promoters and/or 3′-UTRs, isolated from Brachypodium distachyon ubiquitin genes.

Claims (33)

1. A nucleic acid vector comprising a promoter, wherein

i) said promoter is operably linked to a polylinker sequence;

ii) said promoter is operably linked to a non-ubiquitin-coding sequence; or

iii) said promoter is operably linked to a sequence comprising a polylinker sequence and a non-ubiquitin-coding sequence, wherein said promoter comprises SEQ ID NO: 1 or a sequence that has at least 99% sequence identity with SEQ ID NO: 1 and can initiate transcription of an operably linked coding sequence.

2. The nucleic acid vector of claim 1 wherein said promoter is less than 3 kb in length.

3. The nucleic acid vector of claim 1 wherein said promoter consists of SEQ ID NO: 1.

4. The nucleic acid vector of claim 1 further comprising a sequence encoding a selectable marker.

5. The nucleic acid vector of claim 4 wherein said promoter is operably linked to a transgene.

6. The nucleic acid vector of claim 5 wherein the transgene encodes a selectable marker or a gene product conferring insecticidal resistance, herbicide tolerance, nitrogen use efficiency, water use efficiency, or nutritional quality.

7. The nucleic acid vector of claim 1 further comprising a 3′ untranslated sequence comprising SEQ ID NO: 4 or a sequence that has at least 99% sequence identity with SEQ ID NO: 4, wherein the 3′ untranslated sequence is operably linked to said polylinker or said coding sequence.

8. The nucleic acid vector of claim 1 further comprising a 5′ untranslated sequence comprising SEQ ID NO: 11 or a sequence that has at least 99% sequence identity with SEQ ID NO: 11, wherein the 5′ untranslated sequence is inserted between, and operably linked to, said promoter sequence and said polylinker or said coding sequence.

9. The nucleic acid vector of claim 8 further comprising an intron sequence inserted after the 5′ untranslated sequence.

10. The nucleic acid vector of claim 9 wherein the intron sequence comprises SEQ ID NO: 7.

11. The nucleic acid vector of claim 1 wherein the promoter consists of SEQ ID NO: 40 or a sequence having at least 99% sequence identity with SEQ ID NO: 40, and said promoter being operably linked to said polylinker or said coding sequence, wherein said promoter can initiate transcription of the operably linked coding sequence.

12. The nucleic acid vector of claim 1 wherein the promoter consists of SEQ ID NO: 15 or a sequence having at least 99% sequence identity with SEQ ID NO: 15, said promoter being operably linked to said polylinker or said coding sequence, wherein said promoter can initiate transcription of the operably linked coding sequence.

13. The nucleic acid vector of claim 12 further comprising a 3′ untranslated sequence comprising SEQ ID NO: 4 or a sequence that has at least 99% sequence identity with SEQ ID NO: 4, wherein the 3′ untranslated sequence is operably linked to said polylinker or said coding sequence.

14. A non- Brachypodium plant comprising a Brachypodium promoter, wherein said Brachypodium promoter comprises SEQ ID NO: 1, or a sequence that has at least 99% sequence identity with SEQ ID NO: 1 operably linked to a coding sequence wherein said Brachypodium promoter can initiate transcription of the operably linked coding sequence.

15. The plant of claim 14 wherein said plant is selected from the group consisting of maize, wheat, rice, sorghum, oats, rye, bananas, sugar cane, soybean, cotton, sunflower, and canola.

16. The plant of claim 14 wherein said plant is Zea mays.

17. The plant of claim 16 wherein the operably linked coding sequence is a transgene, further wherein said transgene is inserted into the genome of said plant.

18. The plant of claim 14 further comprising a 5′ untranslated sequence comprising SEQ ID NO: 11 or a sequence that has at least 99% sequence identity with SEQ ID NO: 11, wherein the 5′ untranslated sequence is inserted between, and operably linked to, said Brachypodium promoter and said coding sequence.

19. The plant of claim 18 further comprising an intron sequence inserted after the 5′ untranslated sequence.

20. The plant of claim 19 wherein the intron sequence comprises SEQ ID NO: 7.

21. The plant of claim 18 further comprising a 3′ untranslated sequence comprising SEQ ID NO: 4 or a sequence that has at least 99% sequence identity with SEQ ID NO: 4, wherein the 3′ untranslated sequence is operably linked to said coding sequence.

22. The plant of claim 14 wherein the Brachypodium promoter consists of SEQ ID NO: 40 or a sequence having at least 99% sequence identity with SEQ ID NO: 40 and can initiate transcription of the operably linked coding sequence further wherein said Brachypodium promoter is operably linked to said coding sequence.

23. The plant of claim 14 wherein the Brachypodium promoter consists of SEQ ID NO: 15 or a sequence having at least 99% sequence identity with SEQ ID NO: 15 and can initiate transcription of an operably linked coding sequence further wherein said Brachypodium promoter is operably linked to said coding sequence.

24. The plant of claim 23 further comprising a 3′ untranslated sequence comprising SEQ ID NO: 4 or a sequence that has at least 99% sequence identity with SEQ ID NO: 4, wherein the 3′ untranslated sequence is operably linked to said coding sequence.

25. A nucleic acid vector comprising a transcription terminator, wherein

i) said terminator is operably linked to a polylinker sequence;

ii) said terminator is operably linked to a non-ubiquitin-coding sequence; or

iii) said terminator is operably linked to a sequence comprising a polylinker sequence and a non-ubiquitin-coding sequence, wherein said transcription terminator comprises SEQ ID NO: 4 or a sequence that has at least 99% sequence identity with SEQ ID NO: 4.

26. The nucleic acid vector of claim 25 wherein said transcription terminator is less than 1 kb in length.

27. The nucleic acid vector of claim 26 wherein said transcription terminator consists of SEQ ID NO: 4.

Assignments (2)
CHANGE OF NAME Recorded Nov 8, 2021
From: DOW AGROSCIENCES LLC
To: CORTEVA AGRISCIENCE LLC
Reel/Frame 058044/0184 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 13, 2014
From: KUMAR, SANDEEP; GUPTA, MANJU; ALABED, DIAA
To: DOW AGROSCIENCES LLC
Reel/Frame 033937/0705 →