New promoter

Abstract

To provide a novel promoter and a method for producing a target substance using the promoter. [Solution] A promoter-active DNA comprising a nucleotide sequence selected from the group consisting of (a) to (c) below: (a) a nucleotide sequence upstream of a gene encoding a polypeptide consisting of the following amino acid sequence; (b) a nucleotide sequence having at least 90% identity with the sequence in (a); and (c) a nucleotide sequence in which one or more nucleotides are deleted, substituted, added, or inserted from the sequence in (a); MX1X2X3FX1IX4QX5IFX1GX6X1X1LX7X1SX8X1X1GAQX9VFX 10 X1X 11 X1X1X 12 SS(X1 is any amino acid residue, X2 is S or A, X3 is V or I, X4 is F or I, X5 is S or A, X6 is V or I, X7 is V or I, X8 is V or I, X9 is G or N, X 10 is D or T, X 11 is I or V, X 12 (A or F).

Claims

1. DNA having promoter activity, comprising a nucleotide sequence selected from the group consisting of (a) to (c) below and a nucleotide sequence selected from the group consisting of (d) to (f) below: (a) The upstream nucleotide sequence of the gene encoding a polypeptide consisting of the following amino acid sequence; (b) A nucleotide sequence having at least 90% identity with the upstream nucleotide sequence of a gene encoding a polypeptide consisting of the following amino acid sequence; and (c) A nucleotide sequence in which one or more nucleotides are deleted, substituted, added, or inserted into the upstream nucleotide sequence of a gene encoding a polypeptide consisting of the following amino acid sequence; MX １ X ２ X ３ FX １ IX ４ QX ５ IFX １ GX ６ X １ X １ LX ７ X １ SX ８ X １ X １ GAQX ９ VFX １０ X １ X １１ X １ X １ X １２ SS (Here, X １ is any amino acid residue, X ２ is S or A, X ３ is V or I, X ４ is F or I, X ５ is S or A, X ６ is V or I, X ７ is V or I, X ８ is V or I, X ９ is G or N, X １０ is D or T, X １１ is I or V, X １２ (is A or F) (d) A nucleotide sequence that includes a region corresponding to the nucleotide sequence of SEQ ID NO: 21 among the nucleotide sequences of SEQ ID NOs: 23 and 26-41; (e) A nucleotide sequence having at least 95% identity with a nucleotide sequence that includes a region corresponding to the nucleotide sequence of SEQ ID NO: 21 among any of the nucleotide sequences of SEQ ID NOs: 23 and 26-41; and (f) A nucleotide sequence in which 1 to 5 nucleotides are deleted, substituted, added, or inserted into a nucleotide sequence that includes a region corresponding to the nucleotide sequence of sequence number 21 among the nucleotide sequences of sequence numbers 23 and 26 to 41.

2. The DNA according to claim 1, wherein the nucleotide sequence containing the region corresponding to the nucleotide sequence of sequence number 21 among any of the nucleotide sequences of sequence numbers 23 and 26 to 41 is a nucleotide sequence containing the region corresponding to the nucleotide sequence of sequence number 22 among any of the nucleotide sequences of sequence numbers 23 and 26 to 41.

3. The DNA according to claim 1, having promoter activity equivalent to or greater than that of the SPL13 promoter of Sequence ID No.

12.

4. A promoter comprising DNA selected from the group consisting of (g) to (i) below, and DNA selected from the group consisting of (j) to (l) below: (g) DNA consisting of the upstream nucleotide sequence of a gene encoding a polypeptide consisting of the following amino acid sequence; (h) DNA having promoter activity, comprising a nucleotide sequence having at least 90% identity with the upstream nucleotide sequence of a gene encoding a polypeptide consisting of the following amino acid sequence; and (i) DNA having promoter activity, consisting of a nucleotide sequence in which one or more nucleotides are deleted, substituted, added, or inserted into the nucleotide sequence upstream of a gene encoding a polypeptide consisting of the following amino acid sequence; MX １ X ２ X ３ FX １ IX ４ QX ５ IFX １ GX ６ X １ X １ LX ７ X １ SX ８ X １ X １ GAQX ９ VFX １０ X １ X １１ X １ X １ X １２ SS (Here, X １ is any amino acid residue, X ２ is S or A, X ３ is V or I, X ４ is F or I, X ５ is S or A, X ６ is V or I, X ７ is V or I, X ８ is V or I, X ９ is G or N, X １０ is D or T, X １１ is I or V, X １２ (is A or F) (j) DNA consisting of a nucleotide sequence that includes a region corresponding to the nucleotide sequence of SEQ ID NO: 21 among the nucleotide sequences of SEQ ID NOs: 23 and 26-41; (k) DNA having promoter activity, comprising a nucleotide sequence having at least 95% identity with a nucleotide sequence that includes a region corresponding to the nucleotide sequence of SEQ ID NO: 21 among the nucleotide sequences of SEQ ID NOs: 23 and 26-41; and (l) DNA having promoter activity, comprising a nucleotide sequence in which 1 to 5 nucleotides are deleted, substituted, added, or inserted into a nucleotide sequence that includes a region corresponding to the nucleotide sequence of sequence number 21 among the nucleotide sequences of sequence numbers 23 and 26 to 41.

5. The promoter according to claim 4, wherein the nucleotide sequence containing the region corresponding to the nucleotide sequence of SEQ ID NO: 21 among any of the nucleotide sequences of SEQ ID NO: 23 and 26-41 is a nucleotide sequence containing the region corresponding to the nucleotide sequence of SEQ ID NO: 22 among any of the nucleotide sequences of SEQ ID NO: 23 and 26-41.

6. The promoter according to claim 4, having promoter activity equivalent to or greater than that of the SPL13 promoter of Sequence ID No.

12.

7. An expression vector comprising the DNA according to any one of claims 1 to 3 or the promoter according to any one of claims 4 to 6.

8. The expression vector according to claim 7, comprising a gene encoding a target substance or an enzyme involved in its synthesis, and the DNA or promoter ligated upstream of the gene.

9. A DNA expression cassette comprising the DNA according to any one of claims 1 to 3 or the promoter according to any one of claims 4 to 6.

10. The DNA expression cassette according to claim 9, comprising a gene encoding a target substance or an enzyme involved in its synthesis, and the DNA or promoter ligated upstream of the gene.

11. Corynebacterium comprising the expression vector according to claim 7.

12. Corynebacterium comprising the expression vector according to claim 8.

13. Corynebacterium comprising the DNA expression cassette described in claim 9.

14. Corynebacterium comprising the DNA expression cassette according to claim 10.

15. Corynebacterium into which the DNA according to any one of claims 1 to 3 or the promoter according to any one of claims 4 to 6 has been introduced.

16. Corynebacterium according to claim 15, wherein the DNA or promoter is introduced upstream of a gene encoding a target substance or an enzyme involved in its synthesis.

17. Corynebacterium glutamicum as described in claim 11.

18. Corynebacterium glutamicum as described in claim 12.

19. Corynebacterium glutamicum as described in claim 13.

20. Corynebacterium glutamicum as described in claim 14.

21. Corynebacterium glutamicum as described in claim 15.

22. Corynebacterium glutamicum as described in claim 16.

23. Culturing Corynebacterium according to claim 12; and A method for producing a target substance, comprising recovering the target substance from a culture obtained in the culture.

24. Culturing Corynebacterium according to claim 14; and A method for producing a target substance, comprising recovering the target substance from a culture obtained in the culture.

25. Culturing Corynebacterium according to claim 16; and A method for producing a target substance, comprising recovering the target substance from a culture obtained in the culture.

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