Materials and methods for protein production
This document relates to materials and methods for the production of protein. In one aspect, this document provides a nucleic acid construct including a first alcohol oxidase promoter element, wherein the first alcohol oxidase promoter element includes a mutation at one or more nucleotide positions corresponding to any of nucleotide positions 668-734 relative to SEQ ID NO: 28.
1. A nucleic acid construct comprising a first alcohol oxidase promoter element, wherein the first alcohol oxidase promoter element comprises a mutation at one or more nucleotide positions corresponding to any of nucleotide positions 668-734 relative to SEQ ID NO: 28.
2. The nucleic acid construct of claim 1 , wherein the first alcohol oxidase promoter element comprises a mutation at one or more nucleotide positions corresponding to any of nucleotide positions 678-724 relative to SEQ ID NO: 28.
3. The nucleic acid construct of claim 2 , wherein the first alcohol oxidase promoter element comprises a mutation at one or more nucleotide positions corresponding to any of nucleotide positions 688-714 relative to SEQ ID NO: 28.
4. The nucleic acid construct of claim 1 , wherein the first alcohol oxidase promoter element comprises two or more mutations at nucleotide positions corresponding to any of nucleotide positions 668-734 relative to SEQ ID NO: 28.
5. The nucleic acid construct of claim 1 , wherein the first alcohol oxidase promoter element is an alcohol oxidase 1 promoter element.
6. The nucleic acid construct of claim 1 , further comprising a nucleotide sequence, wherein the nucleotide sequence is operably linked to the first alcohol oxidase promoter element.
7. The nucleic acid construct of claim 6 , wherein the nucleotide sequence encodes a first protein.
8. The nucleic acid construct of claim 7 , wherein the first protein is selected from the group consisting of an antibody or fragment thereof, an enzyme, a regulatory protein, a peptide hormone, a blood clotting protein, a cytokine, a cytokine inhibitor, and a heme-binding protein.
9. The nucleic acid construct of claim 7 , wherein the first protein is a heme-binding protein.
10. A cell comprising a first nucleic acid construct, wherein the first nucleic acid construct is the nucleic acid construct of claim 1 .
11. A method of producing a product in a cell comprising:
expressing a nucleic acid construct comprising a nucleotide sequence operably linked to a first alcohol oxidase promoter element, wherein the first alcohol oxidase promoter element comprises a mutation at one or more nucleotide positions corresponding to any of nucleotide positions 668-734 relative to SEQ ID NO: 28.
12. The method of claim 11 , wherein the first alcohol oxidase promoter element comprises a mutation at one or more nucleotide positions corresponding to any of nucleotide positions 678-724 relative to SEQ ID NO: 28.
13. The method of claim 11 , wherein the first alcohol oxidase promoter element is an alcohol oxidase 1 promoter element.
14. The method of claim 11 , wherein the nucleotide sequence operably linked to the first alcohol oxidase promoter element encodes a first protein.
15. The method of claim 11 , wherein the method is carried out in the absence of added methanol.
16. The method of claim 11 , wherein the method is carried out in the presence of added methanol.
17. The method of claim 11 , wherein the first alcohol oxidase promoter element comprises a mutation at one or more nucleotide positions corresponding to any of nucleotide positions 688-714 relative to SEQ ID NO: 28.
18. The method of claim 11 , wherein the first alcohol oxidase promoter element comprises two or more mutations at nucleotide positions corresponding to any of nucleotide positions 668-734 relative to SEQ ID NO: 28.
19. The method of claim 14 , wherein the first protein is selected from the group consisting of an antibody or fragment thereof, an enzyme, a regulatory protein, a peptide hormone, a blood clotting protein, a cytokine, a cytokine inhibitor, and a heme-binding protein.
20. The method of claim 14 , wherein the first protein is a heme-binding protein.