IP Library Granted Patent US 10,266,833
Granted Patent B2
US 10,266,833 · App. 15/745,070 · Granted Apr 23, 2019

Nucleic acid sequence for protein expression

Inventors: Satoru Shinkawa (Saitama, JP); Shigenobu Mitsuzawa (Saitama, JP); Maiko Tanaka (Saitama, JP); Masayuki Machida (Hokkaido, JP); Hiroko Hagiwara (Ibaraki, JP); Hideaki Koike (Ibaraki, JP); Mahoko Kuninaga (Ibaraki, JP)
Assignee: Honda Motor Co., Ltd.
C12N15/80C07K14/38C12N9/2402C12N15/63C12Y302/01031
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Quick Facts
Patent No.
US 10,266,833
App. No.
15/745,070
Granted
Apr 23, 2019
Kind
B2
Abstract

To provide a base sequence for protein expression that can increase the yield of protein such as diastatic enzyme by further activating a promoter of a particular gene. A base sequence 1 for protein expression includes: a gene 3 encoding protein 2 ; a promoter 4 of the gene 3 , the promoter being linked upstream of the gene 3 ; and a cis element 5 whose activity is improved by an artificial transcription factor 6 , the cis element being linked further upstream of the promoter 4 . The cis element 5 is represented by SEQ ID NO: 1.

Claims (29)

1. A nucleic acid sequence for protein expression comprising:

a gene encoding a protein to be expressed;

a promoter for the gene, the promoter being linked upstream of the gene; and

a cis element comprising the nucleic acid sequence of SEQ ID NO: 1, whose activity is improved by an artificial transcription factor, the cis element being linked further upstream of the promoter, wherein the artificial transcription factor is encoded by the nucleic acid of SEQ ID NO: 2.

2. The nucleic acid sequence for protein expression according to claim 1 , wherein the artificial transcription factor is encoded by the nucleic acid sequence of SEQ ID NO: 2.

3. The nucleic acid sequence for protein expression according to claim 2 , wherein the nucleic acid sequence for protein expression comprises a nucleic acid sequence for the artificial transcription factor to be expressed comprising: a gene encoding the artificial transcription factor encoded by the nucleic acid sequence of SEQ ID NO: 2; and a promoter for the gene encoding the artificial transcription factor, the promoter being linked upstream of the gene encoding the artificial transcription factor.

4. The nucleic acid sequence for protein expression according to claim 1 , wherein the cis element with any number selected in a range of 1 to 10 is/are linked upstream of the promoter of the gene encoding the protein.

5. An expression vector including a nucleic acid sequence for protein expression, the nucleic acid sequence for protein expression comprising:

a gene encoding a protein to be expressed;

a promoter for the gene, the promoter being linked upstream of the gene; and

a cis element comprising the nucleic acid sequence of SEQ ID NO: 1, whose activity is improved by an artificial transcription factor, the cis element being linked further upstream of the promoter, wherein the artificial transcription factor is encoded by the nucleic acid of SEQ ID NO: 2.

6. The expression vector according to claim 5 , wherein the artificial transcription factor is encoded by the nucleic acid sequence of SEQ ID NO: 2.

7. The expression vector according to claim 6 , wherein the nucleic acid sequence for protein expression comprises a nucleic acid sequence for the artificial transcription factor to be expressed comprising: a gene encoding the artificial transcription factor encoded by the nucleic acid sequence of SEQ ID NO: 2; and a promoter for the gene encoding the artificial transcription factor, the promoter being linked upstream of the gene encoding the artificial transcription factor.

8. The expression vector according to claim 5 , wherein in the nucleic acid sequence for protein expression, the cis element with any number selected in a range of 1 to 10 is/are linked upstream of the promoter of the gene encoding the protein.

9. A transformant including a nucleic acid sequence for protein expression, the nucleic acid sequence for protein expression comprising:

a gene encoding a protein to be expressed;

a promoter for the gene, the promoter being linked upstream of the gene; and

a cis element comprising the nucleic acid sequence of SEQ ID NO: 1, whose activity is improved by an artificial transcription factor, the cis element being linked further upstream of the promoter, wherein the artificial transcription factor is encoded by the nucleic acid of SEQ ID NO: 2.

10. The transformant according to claim 9 , wherein the artificial transcription factor is encoded by the nucleic acid sequence of SEQ ID NO: 2.

11. The transformant according to claim 10 , wherein the nucleic acid sequence for protein expression comprises a nucleic acid sequence for the artificial transcription factor to be expressed comprising: a gene encoding the artificial transcription factor encoded by the nucleic acid sequence of SEQ ID NO: 2; and a promoter for the gene encoding the artificial transcription factor, the promoter being linked upstream of the gene encoding the artificial transcription factor.

12. The transformant according to claim 9 , wherein in the nucleic acid sequence for protein expression, the cis element with any number selected in a range of 1 to 10 is/are linked upstream of the promoter of the gene encoding the protein.

13. The transformant according to claim 9 , wherein koji mold is used as a host cell.

14. The transformant according to claim 13 , wherein the koji mold is an Aspergillus oryzae HO2strain (National Institute of Technology and Evaluation, Patent Microorganisms Depositary, #122, 2-5-8 Kazusakamatari, Kisarazu-shi, Chiba, Japan, Deposition Date: Nov. 12, 2013, Deposition No.: NITE BP-01750).

15. The nucleic acid sequence for protein expression according to claim 1 , wherein the nucleic acid sequence for protein expression comprises a nucleic acid sequence for the artificial transcription factor to be expressed comprising: a gene encoding the artificial transcription factor; and a promoter for the gene encoding the artificial transcription factor, the promoter being linked upstream of the gene encoding the artificial transcription factor.

16. The nucleic acid sequence for protein expression according to claim 2 , wherein the cis element with any number selected in a range of 1 to 10 is/are linked upstream of the promoter of the gene encoding the protein.

17. The expression vector according to claim 5 , wherein the nucleic acid sequence for protein expression comprises a nucleic acid sequence for the artificial transcription factor to be expressed comprising: a gene encoding the artificial transcription factor; and a promoter for the gene encoding the artificial transcription factor, the promoter being linked upstream of the gene encoding the artificial transcription factor.

18. The expression vector according to claim 6 , wherein in the nucleic acid sequence for protein expression, the cis element with any number selected in a range of 1 to 10 is/are linked upstream of the promoter of the gene encoding the protein.

19. The transformant according to claim 9 , wherein the nucleic acid sequence for protein expression comprises a nucleic acid sequence for the artificial transcription factor to be expressed comprising: a gene encoding the artificial transcription factor; and a promoter for the gene encoding the artificial transcription factor, the promoter being linked upstream of the gene encoding the artificial transcription factor.

20. The transformant according to claim 10 , wherein in the nucleic acid sequence for protein expression, the cis element with any number selected in a range of 1 to 10 is/are linked upstream of the promoter of the gene encoding the protein.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 25, 2018
From: SHINKAWA, SATORU; MITSUZAWA, SHIGENOBU; TANAKA, MAIKO; MACHIDA, MASAYUKI; HAGIWARA, HIROKO; KOIKE, HIDEAKI; KUNINAGA, MAHOKO
To: HONDA MOTOR CO., LTD.
Reel/Frame 044727/0275 →
Priority Claims (1)
JP 2015-142213 · Jul 16, 2015 · national
Continuity (1)
Related Publication 20180201942A1 · Jul 19, 2018