Plant terminator sequences
This invention relates to gene expression regulatory sequences, specifically transcription terminator sequences. Plant transcription terminator sequences are described herein. Methods for identifying novel plant transcription terminator sequences that can terminate transcription in one orientation or in a bidirectional manner and methods of using these terminator sequences to generate transgenic plants are described herein.
1. A recombinant construct comprising a polynucleotide comprising:
(a) a nucleotide sequence as set forth in SEQ ID NO 3 or 10; or
(b) a functional fragment of at least 200 contiguous nucleotides of (a) having terminator activity;
wherein the polynucleotide is operably linked to the 3′ end of a heterologous polynucleotide, wherein the heterologous polynucleotide is operably linked to a promoter functional in a plant cell; and further wherein the polynucleotide functions as a transcriptional terminator for the heterologous polynucleotide in a plant cell.
2. The recombinant construct of claim 1 wherein the polynucleotide functions as a bidirectional transcriptional terminator.
3. The recombinant construct of claim 2 wherein the bidirectional transcriptional terminator is operably linked to:
(a) the 3′ end of a first heterologous polynucleotide, wherein the heterologous polynucleotide is operably linked to a first promoter functional in a plant cell; and
(b) the 3′ end of a second heterologous polynucleotide, wherein the second heterologous polynucleotide which is operably linked to a second promoter functional in a plant cell;
wherein the first and the second heterologous polynucleotides are transcribed in a convergent manner.
4. A plant comprising in its genome the recombinant construct of claim 1 .
5. A seed from the plant of claim 4 , wherein the seed comprises said recombinant construct.
6. The plant of claim 4 , wherein said plant is selected from the group consisting of: Arabidopsis , maize, soybean, sunflower, sorghum, canola, mustard, wheat, alfalfa, cotton, rice, barley, millet, sugar cane and switchgrass.
7. The seed of claim 5 , wherein said seed is selected from the group consisting of: Arabidopsis , maize, soybean, sunflower, sorghum, canola, mustard, wheat, alfalfa, cotton, rice, barley, millet, sugar cane and switchgrass.
8. A method of expressing a heterologous polynucleotide in a plant, the method comprising the steps of:
(a) introducing into a regenerable plant cell the recombinant construct of claim 1 ;
(b) regenerating a transgenic plant from the regenerable plant cell of (a), wherein the transgenic plant comprises in its genome the recombinant construct of claim 1 ; and
(c) obtaining a progeny plant from the transgenic plant of step (b), wherein the progeny plant comprises in its genome the recombinant construct of claim 1 and exhibits expression of the heterologous polynucleotide.
9. A method of regulating the expression of two heterologous polynucleotides in a plant, comprising the steps of:
(a) introducing into a regenerable plant cell the recombinant construct of claim 3 ;
(b) regenerating a transgenic plant from the regenerable plant cell of (a), wherein the transgenic plant comprises in its genome the recombinant construct of claim 3 ; and
(c) obtaining a progeny plant from the transgenic plant of step (b), wherein the progeny plant comprises in its genome the recombinant construct of claim 3 and exhibits expression of both the first heterologous polynucleotide and the second heterologous polynucleotide.
10. The method of claim 8 , wherein said plant is selected from the group consisting of: Arabidopsis , maize, soybean, sunflower, sorghum, canola, mustard, wheat, alfalfa, cotton, rice, barley, millet, sugar cane and switchgrass.
11. The method of claim 9 , wherein said plant is selected from the group consisting of: Arabidopsis , maize, soybean, sunflower, sorghum, canola, mustard, wheat, alfalfa, cotton, rice, barley, millet, sugar cane and switchgrass.
12. A plant comprising in its genome the recombinant construct of claim 3 .
13. A seed from the plant of claim 12 , wherein the seed comprises said recombinant construct.
14. The plant of claim 12 , wherein said plant is selected from the group consisting of: Arabidopsis , maize, soybean, sunflower, sorghum, canola, mustard, wheat, alfalfa, cotton, rice, barley, millet, sugar cane and switchgrass.
15. The seed of claim 13 , wherein said seed is selected from the group consisting of: Arabidopsis , maize, soybean, sunflower, sorghum, canola, mustard, wheat, alfalfa, cotton, rice, barley, millet, sugar cane and switchgrass.