Synthetic chloroplast transit peptides
This disclosure concerns compositions and methods for targeting peptides, polypeptides, and proteins to plastids of plastid-containing cells. In some embodiments, the disclosure concerns chloroplast transit peptides that may direct a polypeptide to a plastid, and nucleic acid molecules encoding the same. In some embodiments, the disclosure concerns methods for producing a transgenic plant material (e.g., a transgenic plant) comprising a chloroplast transit peptide, as well as plant materials produced by such methods, and plant commodity products produced therefrom.
1. An isolated nucleic acid molecule comprising a polynucleotide, wherein the polynucleotide comprises an EPSPS-derived means for targeting a polypeptide to a chloroplast, and a nucleotide sequence encoding a polypeptide of interest operably linked in frame in the polynucleotide to the EPSPS-derived means for targeting a polypeptide to a chloroplast,
wherein the polynucleotide encodes a chimeric protein comprising the polypeptide of interest and a peptide encoded by the EPSPS-derived means for targeting a polypeptide to a chloroplast,
wherein the EPSPS-derived means for targeting a polypeptide to a chloroplast encodes a peptide consisting of an amino acid sequence that is at least 80% identical to SEQ ID NO: 11, and
wherein the peptide encoded by the EPSPS-derived means for targeting a polypeptide to a chloroplast is not a native plant chloroplast transit peptide (CTP).
2. The isolated nucleic acid molecule of claim 1 , wherein the EPSPS-derived means for targeting a polypeptide to a chloroplast encodes a peptide consisting of an amino acid sequence that is at least 90% identical to SEQ ID NO: 11.
3. The isolated nucleic acid molecule of claim 1 , wherein the EPSPS-derived means for targeting a polypeptide to a chloroplast encodes a peptide consisting of an amino acid sequence that is at least 95% identical to SEQ ID NO: 11.
4. The nucleic acid molecule of claim 1 , wherein the molecule is a plant expression vector.
5. The chimeric protein encoded by the polynucleotide of the nucleic acid molecule of claim 1 .
6. The chimeric protein of claim 5 , wherein the polypeptide of interest is an herbicide tolerance gene product or insect resistance gene product.
7. A transgenic plant material comprising the nucleic acid molecule of claim 1 .
8. The transgenic plant material of claim 1 , wherein the plant material is selected from the group consisting of a plant cell, a plant tissue, a plant tissue culture, a callus culture, and a plant part.
9. The transgenic plant material of claim 8 , wherein the polynucleotide is stably integrated into the genome of a cell from the plant material.
10. The transgenic plant material of claim 7 , wherein the plant material is a whole plant, and wherein the polynucleotide is stably integrated into the genome of the plant.
11. A method for producing a transgenic plant material, the method comprising transforming a plant material with the nucleic acid molecule of claim 1 .
12. A method for producing a transgenic plant, the method comprising regenerating a transgenic plant from the transgenic plant material of claim 9 .
13. The chimeric protein of claim 6 , wherein the polypeptide of interest is a bacterial 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS).
14. The chimeric protein of claim 13 , wherein the polypeptide of interest is DGT-28.
15. The chimeric protein of claim 6 , wherein the polypeptide of interest is an insecticidal bacterial protein.
16. The chimeric protein of claim 15 , wherein the polypeptide of interest is Cry2Aa.