Promoters from
Provided are native promoters comprising polynucleotides isolated from Corynebacterium glutamicum , and mutant promoters derived therefrom, which may be used to regulate, i.e., either increase or decrease, gene expression. Also provided are promoter ladders comprising a plurality of the promoters having incrementally increasing promoter activity. Also provided are host cells and recombinant vectors comprising the promoters, and methods of expressing genes of interest and producing biomolecules using the host cells.
1. A promoter ladder comprising at least three promoter polynucleotide sequences selected from the group consisting of SEQ ID NOs: 1-8, wherein said promoter ladder comprises a plurality of promoters with incrementally increasing levels of promoter activity.
2. The promoter ladder according to claim 1 , wherein at least one of said promoter polynucleotide sequences is selected from the group consisting of SEQ ID NOs: 1, 5 and 7.
3. The promotor ladder according to claim 1 , wherein at least one of said promoter polynucleotide sequences is selected from the group consisting of SEQ ID NOs: 1-2.
4. The promoter ladder according to claim 1 , wherein at least one of said promoter polynucleotide sequences is selected from the group consisting of SEQ ID NOs: 3, 4 and 5.
5. The promotor ladder according to claim 1 , wherein at least one of said promoter polynucleotide sequences is selected from the group consisting of SEQ ID NOs: 6-8.
6. The promoter ladder according to claim 1 , wherein each promoter of said promoter ladder is functionally linked to at least one heterologous target gene.
7. The promotor ladder according to claim 6 , wherein said at least one heterologous target gene is a gene that is a component of a biosynthetic pathway producing a biomolecule selected from the group consisting of amino acids, organic acids, proteins and polymers.
8. The promoter ladder accordingly to claim 7 , wherein said at least one heterologous target gene is a gene that is selected from the group consisting of: aspartate-semialdehyde dehydrogenase, 4-hydroxy-tetrahydrodipicolinate synthase, dihydrodipicolinate reductase, 2,3,4,5-tetrahydropyridine-2-carboxylate N-succinyltransferase, N-succinyldiaminopimelate aminotransferase, succinyl-diaminopimelate desuccinylase, diaminopimelate epimerase, diaminopimelate decarboxylase, diaminopimelate dehydrogenase, aspartokinase lysc alpha and beta subunits, aspartate aminotransferase, glucose-specific enzyme II BC component of phosphotransferase System (PTS), glucose-6-phosphate 1-dehydrogenase, glucose-6-phosphate isomerase, transketolase, 6-phosphofructokinase 1, phosphoenolpyruvate carboxylase, pyruvate carboxylase, isocitrate dehydrogenase, phosphoenolpyruvate carboxykinase (GTP), oxaloacetate decarboxylase, homoserine kinase, homoserine dehydrogenase, and threonine synthase.
9. A recombinant polynucleotide comprising the promoter ladder of claim 1 functionally linked to a heterologous target gene.
10. A plurality of recombinant polynucleotides comprising the promoter ladder of claim 1 , wherein each recombinant polynucleotide comprises one promoter from the promoter ladder functionally linked to a heterologous target gene.
11. A plurality if recombinant vectors comprising the promoter ladder of claim 1 , the recombinant polynucleotide of claim 9 , or the plurality of recombinant polynucleotides of claim 10 .
12. A plurality of recombinant vectors comprising the promoter ladder of claim 1 , the recombinant polynucleotide of claim 9 , or the plurality of recombinant polynucleotides of claim 10 .
13. The plurality of host cells according to claim 12 , wherein the host cells belong to the genus Corynebacterium.
14. A plurality of transformed host cells comprising a combination of at least three promoter polynucleotide sequences selected from the group consisting of SEQ ID NOs: 1-8 each functionally linked to at least one heterologous target gene, wherein said combination of promoter polynucleotide sequences comprises a plurality of promoters with incrementally increasing levels of promoter activity.
15. The plurality of transformed host cells according to claim 14 , wherein said target gene is associated with a biosynthetic pathway producing a biomolecule selected from: amino acids, organic acids, flavors and fragrances, biofuels, proteins and enzymes, polymers/monomers and other biomaterials, lipids, nucleic acids, small molecule therapeutics, protein therapeutics, fine chemicals, and nutraceuticals.
16. The plurality of transformed host cells according to claim 14 , wherein said target gene is associated with a biosynthetic pathway producing a secondary metabolite selected from: antibiotics, alkaloids, terpenoids, and polyketides.
17. The plurality of transformed host cells according to claim 14 , wherein each promoter polynucleotide sequence is functionally linked to a different heterologous target gene.
18. The plurality of transformed host cells according to claim 14 , wherein two or more promoter polynucleotide sequences are functionally linked to at least one heterologous target gene.
19. The plurality of transformed host cells according to claim 18 , wherein the two or more promoter polynucleotide sequences are the same.
20. The plurality of transformed host cells according to claim 18 , wherein the two or more promoter polynucleotide sequences are different.