Increased protein expression in plants
This disclosure concerns synthetic polynucleotides encoding a polypeptide of interest that are particularly well-suited for expression in target plants.
1. A method of controlling pests, the method comprising obtaining grain from a transgenic maize plant, wherein the transgenic maize plant contains a synthetic polynucleotide comprising:
a nucleotide sequence encoding the polypeptide of interest, wherein the nucleotide sequence has been codon-optimized to remove codons selected from the group consisting of TTA, CTA, GTA, CGT, AGT, and CGA from the reference nucleotide,
wherein the synthetic polynucleotide nucleotide sequence encoding the polypeptide of interest comprises at least one polyadenylation sequence selected from the group consisting of AATAAA, AATAAT, AACCAA, ATATAA, AATCAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA, present in the same number and in the same location as in the reference nucleotide sequence,
wherein the synthetic polynucleotide nucleotide sequence encoding the polypeptide of interest does not comprise any polyadenylation sequence selected from the group consisting of AATAAT, AACCAA, ATATAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA in a different number or location than in the reference nucleotide sequence,
wherein the synthetic polynucleotide nucleotide sequence encoding the polypeptide of interest does not comprise a polyadenylation sequence selected from the group consisting of ATATAT, TTGTTT, TTTTGT, TGTTTT, TATATA, TATTIT, TTTTTT, ATTTTT, TTATTT, TTTATT, TAATAA, ATTTAT, TATATT, TITTAT, ATATTT, TATTAT, TGTTTG, TTATAT, TGTAAT, AAATAA, AATTTT, TTTTTA, TAATTT, TTAATT, AAATTT, TTTGTT, ATTATT, ATTTTA, TITTAAT, and TTTTAA,
wherein the synthetic polynucleotide comprises a 5′ non-translated sequence and a 3′ non-translated region, wherein the 5′ non-translated sequence comprises a plant promoter operably linked to the sequence encoding the polypeptide of interest, and
wherein the 3′ non-translated sequence comprises a transcription termination sequence.
2. A method of controlling pests, the method comprising obtaining seed from a transgenic maize plant, wherein the transgenic maize plant contains a synthetic polynucleotide comprising:
a nucleotide sequence encoding the polypeptide of interest, wherein the nucleotide sequence has been codon-optimized to remove codons selected from the group consisting of TTA, CTA, GTA, CGT, AGT, and CGA from the reference nucleotide,
wherein the synthetic polynucleotide nucleotide sequence encoding the polypeptide of interest comprises at least one polyadenylation sequence selected from the group consisting of AATAAA, AATAAT, AACCAA, ATATAA, AATCAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA, present in the same number and in the same location as in the reference nucleotide sequence,
wherein the synthetic polynucleotide nucleotide sequence encoding the polypeptide of interest does not comprise any polyadenylation sequence selected from the group consisting of AATAAT, AACCAA, ATATAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA in a different number or location than in the reference nucleotide sequence,
wherein the synthetic polynucleotide nucleotide sequence encoding the polypeptide of interest does not comprise a polyadenylation sequence selected from the group consisting of ATATAT, TTGTTT, TITTGT, TGTTTT, TATATA, TATTTT, TTTT ATTTTT, TTATTT, TTTATT, TAATAA, ATTTAT, TATATT, TITTAT, ATATIT, TATTAT, TGTTTG, TTATAT, TGTAAT, AAATAA, AATTTT, TTTTTA, TAATTT, TTAATT, AAATTT, TITGTT, ATTATT, ATTTTA, TTTAAT, and TITTAA,
wherein the synthetic polynucleotide comprises a 5′ non-translated sequence and a 3′ non-translated region, wherein the 5′ non-translated sequence comprises a plant promoter operably linked to the sequence encoding the polypeptide of interest, and
wherein the 3′ non-translated sequence comprises a transcription termination sequence.
3. The method according to claim 2 , wherein the method further comprises germinating the seed.