JPWO2019156152A5 - - Google Patents
Download PDFInfo
- Publication number
- JPWO2019156152A5 JPWO2019156152A5 JP2019570794A JP2019570794A JPWO2019156152A5 JP WO2019156152 A5 JPWO2019156152 A5 JP WO2019156152A5 JP 2019570794 A JP2019570794 A JP 2019570794A JP 2019570794 A JP2019570794 A JP 2019570794A JP WO2019156152 A5 JPWO2019156152 A5 JP WO2019156152A5
- Authority
- JP
- Japan
- Prior art keywords
- nucleotide sequence
- nucleic acid
- promoter
- arabinose
- sequence
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000002773 nucleotide Substances 0.000 claims 52
- 125000003729 nucleotide group Chemical group 0.000 claims 52
- 108020004707 nucleic acids Proteins 0.000 claims 33
- 102000039446 nucleic acids Human genes 0.000 claims 33
- 150000007523 nucleic acids Chemical class 0.000 claims 33
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 claims 22
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims 22
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims 22
- 230000014509 gene expression Effects 0.000 claims 16
- 230000000694 effects Effects 0.000 claims 14
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 claims 12
- 241000186254 coryneform bacterium Species 0.000 claims 11
- 241000894006 Bacteria Species 0.000 claims 10
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 claims 8
- 241000186031 Corynebacteriaceae Species 0.000 claims 7
- 108090000623 proteins and genes Proteins 0.000 claims 7
- 229940113082 thymine Drugs 0.000 claims 6
- 108020004414 DNA Proteins 0.000 claims 5
- 239000013600 plasmid vector Substances 0.000 claims 5
- 101150076178 araE gene Proteins 0.000 claims 4
- 101150055570 araR gene Proteins 0.000 claims 4
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 claims 4
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 claims 4
- 229940035893 uracil Drugs 0.000 claims 4
- 229930024421 Adenine Natural products 0.000 claims 2
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims 2
- 108091028043 Nucleic acid sequence Proteins 0.000 claims 2
- 229960000643 adenine Drugs 0.000 claims 2
- 229940104302 cytosine Drugs 0.000 claims 2
- 239000013612 plasmid Substances 0.000 claims 2
- 230000010076 replication Effects 0.000 claims 2
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 claims 1
- 101100465553 Dictyostelium discoideum psmB6 gene Proteins 0.000 claims 1
- 102100033238 Elongation factor Tu, mitochondrial Human genes 0.000 claims 1
- 241000588722 Escherichia Species 0.000 claims 1
- 101000851240 Homo sapiens Elongation factor Tu, mitochondrial Proteins 0.000 claims 1
- 101100425947 Mus musculus Tnfrsf13b gene Proteins 0.000 claims 1
- 101100169519 Pyrococcus abyssi (strain GE5 / Orsay) dapAL gene Proteins 0.000 claims 1
- 101100398785 Streptococcus agalactiae serotype V (strain ATCC BAA-611 / 2603 V/R) ldhD gene Proteins 0.000 claims 1
- 101100386830 Zymomonas mobilis subsp. mobilis (strain ATCC 31821 / ZM4 / CP4) ddh gene Proteins 0.000 claims 1
- 238000010367 cloning Methods 0.000 claims 1
- 230000000295 complement effect Effects 0.000 claims 1
- 230000001276 controlling effect Effects 0.000 claims 1
- 101150011371 dapA gene Proteins 0.000 claims 1
- 239000013604 expression vector Substances 0.000 claims 1
- 101150026107 ldh1 gene Proteins 0.000 claims 1
- 101150041530 ldha gene Proteins 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 239000003550 marker Substances 0.000 claims 1
- 238000000034 method Methods 0.000 claims 1
- 230000001105 regulatory effect Effects 0.000 claims 1
- 108091008146 restriction endonucleases Proteins 0.000 claims 1
- 239000013076 target substance Substances 0.000 claims 1
- 238000011144 upstream manufacturing Methods 0.000 claims 1
- 239000013598 vector Substances 0.000 claims 1
- 108700026220 vif Genes Proteins 0.000 claims 1
Claims (31)
以下の（ａ）又は（ｂ）に記載のヌクレオチド配列（Ｙ）を含む、核酸：
（ａ）配列番号５から９、１１及び１２並びに配列番号１７及び１８の何れか１つに記載のヌクレオチド配列；
（ｂ）上記（ａ）に記載のヌクレオチド配列において１又は複数の塩基が欠失、置換若しくは付加されたヌクレオチド配列、
但し、
ヌクレオチド配列（Ｙ）が、以下の一般式（Ｉ）で表され、
５’－Ｎ １ ＴＧＴＮ ２ ＡＧＣＧＮ ３ ＴＮ ４ ＡＮ ５ Ｎ ６ Ｎ ７ －３’・・・一般式（Ｉ）
式中、Ｎ １ 、Ｎ ２ 、Ｎ ３ 、Ｎ ４ 、Ｎ ５ 、Ｎ ６ 及びＮ ７ は、それぞれ、独立にＡ（アデニン）、Ｇ（グアニン）、Ｃ（シトシン）若しくはＴ（チミン）を示し又は欠失するものとし、
核酸がＲＮＡの場合は、ヌクレオチド配列中のチミン（ｔ）をウラシル（ｕ）に読み替えるものとし、
ヌクレオチド配列（Ｙ）として配列番号１０、２０及び２１それぞれに示されるヌクレオチド配列は除外され、かつヌクレオチド配列（Ｙ）は以下の条件（ＩＩ）を満たすものとする、
条件（ＩＩ）：配列番号８０に示すプラスミドベクターｐＧＥ７２８－１のヌクレオチド配列における第２９１３番目から第２９２８番目までのヌクレオチド配列を上記ヌクレオチド配列（Ｙ）と入れ替えたプラスミドベクターで形質転換されたコリネ型細菌において、アラビノースを添加した場合に対してアラビノース未添加の場合にＰｔａｃＩのプロモーター活性が抑制される。 A recombinant nucleic acid for use in controlling the expression of a target gene in coryneform bacteria.
Nucleic acid comprising the nucleotide sequence (Y) according to the following (a) or (b):
(A) The nucleotide sequence according to any one of SEQ ID NOs: 5 to 9, 11 and 12 and SEQ ID NOs: 17 and 18;
(B) A nucleotide sequence in which one or more bases are deleted, substituted or added in the nucleotide sequence described in (a) above.
however,
The nucleotide sequence (Y) is represented by the following general formula (I).
5'-N 1 TGTN 2 AGCGN 3 TN 4 AN 5 N 6 N 7 -3'... General formula (I)
In the formula, N 1 , N 2 , N 3 , N 4 , N 5 , N 6 and N 7 independently indicate A (adenine), G (guanine), C (cytosine) or T (thymine), respectively. Or it shall be deleted
When the nucleic acid is RNA, thymine (t) in the nucleotide sequence shall be read as uracil (u).
The nucleotide sequence shown in SEQ ID NOs: 10, 20 and 21 as the nucleotide sequence (Y) is excluded, and the nucleotide sequence (Y) satisfies the following condition (II).
Condition (II): Corine-type bacterium transformed with a plasmid vector in which the nucleotide sequence from the 2913th to the 2928th in the nucleotide sequence of the plasmid vector pGE728-1 shown in SEQ ID NO: 80 was replaced with the above nucleotide sequence (Y). In the case where arabinose is added, the promoter activity of PtacI is suppressed when arabinose is not added.
上記遺伝子発現制御配列（Ｒ）は、天然には存在しないヌクレオチド配列を有し、
遺伝子発現制御配列（Ｒ）が以下の条件（Ｉ）を充足する、核酸：
（ａ）配列番号５から１２並びに配列番号１７及び１８の何れか１つに記載のヌクレオチド配列；
（ｂ）上記（ａ）に記載のヌクレオチド配列において１又は複数の塩基が欠失、置換若しくは付加されたヌクレオチド配列、
但し、核酸がＲＮＡの場合は、ヌクレオチド配列中のチミン（ｔ）をウラシル（ｕ）に読み替えるものとする、
条件（Ｉ）：上記遺伝子発現制御配列（Ｒ）を含む核酸が導入されたコリネ型細菌において、アラビノース添加の場合に対し、アラビノース未添加の場合に上記ヌクレオチド配列（Ｘ）のプロモーター活性が上記ヌクレオチド配列（Ｙ）の存在により抑制されること。 A gene expression control sequence (R) having at least a part of the nucleotide sequence (X) capable of functioning as a promoter and the nucleotide sequence (Y) described in the following (a) or (b) is included.
The gene expression control sequence (R) has a nucleotide sequence that does not exist in nature and has a nucleotide sequence.
Nucleic acid: The gene expression control sequence (R) satisfies the following condition (I).
(A) The nucleotide sequence according to any one of SEQ ID NOs: 5 to 12 and SEQ ID NOs: 17 and 18;
(B) A nucleotide sequence in which one or more bases are deleted, substituted or added in the nucleotide sequence described in (a) above.
However, when the nucleic acid is RNA, thymine (t) in the nucleotide sequence shall be read as uracil (u).
Condition (I): In a coryneform bacterium into which a nucleic acid containing the gene expression control sequence (R) has been introduced, the promoter activity of the nucleotide sequence (X) is higher than that of the nucleotide in the case of adding arabinose to the case of not adding arabinose. Suppressed by the presence of sequence (Y) .
５’－Ｎ１ＴＧＴＮ２ＡＧＣＧＮ３ＴＮ４ＡＮ５Ｎ６Ｎ７－３’・・・一般式（Ｉ）
式中、Ｎ１、Ｎ２、Ｎ３、Ｎ４、Ｎ５、Ｎ６及びＮ７は、それぞれ、独立にＡ（アデニン）、Ｇ（グアニン）、Ｃ（シトシン）若しくはＴ（チミン）を示し又は欠失し、核酸がＲＮＡの場合は、Ｔ（チミン）はＵ（ウラシル）と読み替えるものとする。 The nucleic acid according to claim 2 , wherein the nucleotide sequence (Y) is represented by the following general formula (I):
5'-N 1 TGTN 2 AGCGN 3 TN 4 AN 5 N 6 N 7 -3'... General formula (I)
In the formula, N 1 , N 2 , N 3 , N 4 , N 5 , N 6 and N 7 independently indicate A (adenine), G (guanine), C (cytosine) or T (thymine), respectively. Or if it is deleted and the nucleic acid is RNA, T (thymine) shall be read as U (uracil).
（ｌ）配列番号２２から６１並びに配列番号９６から１０２の何れか１つに記載のヌクレオチド配列；
（ｍ）（ｌ）に記載のヌクレオチド配列において１又は複数の塩基が欠失、置換若しくは付加されたヌクレオチド配列；
（ｎ）（ｌ）に記載のヌクレオチド配列に対して相補であるヌクレオチド配列とストリンジェントな条件でハイブリダイズするヌクレオチド配列；及び
（ｏ）（ｌ）に記載のヌクレオチド配列と少なくとも８０％以上の配列同一性を有するヌクレオチド配列、
但し、核酸がＲＮＡの場合は、塩基配列中のチミン（ｔ）をウラシル（ｕ）に読み替えるものとする。 The nucleic acid according to any one of claims 2 to 13 , wherein the gene expression control sequence (R) contains any of the following nucleotide sequences (l) to (o).
(L) The nucleotide sequence according to any one of SEQ ID NOs: 22 to 61 and SEQ ID NOs: 96 to 102;
(M) A nucleotide sequence in which one or more bases are deleted, substituted or added in the nucleotide sequence described in (l);
(N) A nucleotide sequence that hybridizes with a nucleotide sequence complementary to the nucleotide sequence described in (l) under stringent conditions; and a sequence of at least 80% or more of the nucleotide sequence described in (o) (l). Nucleotide sequence with identity,
However, when the nucleic acid is RNA, thymine (t) in the base sequence shall be read as uracil (u).
（ａ－１）配列番号６、７及び１７の何れか１つに記載のヌクレオチド配列；(A-1) The nucleotide sequence according to any one of SEQ ID NOs: 6, 7 and 17;
（ｂ－１）上記（ａ－１）に記載のヌクレオチド配列において１又は複数の塩基が欠失、置換若しくは付加されたヌクレオチド配列、(B-1) A nucleotide sequence in which one or more bases are deleted, substituted or added in the nucleotide sequence described in (a-1) above.
条件（ＩＩＩ）配列番号８０に示すプラスミドベクターｐＧＥ７２８－１のヌクレオチド配列における第２９１３番目から第２９２８番目までのヌクレオチド配列を上記ヌクレオチド配列（Ｙ）と入れ替えたプラスミドベクターで形質転換されたコリネ型細菌（Ｓ）において、アラビノースを添加した場合のＰｔａｃＩプロモーター活性（Ａ）に対し、アラビノース未添加の場合のＰｔａｃＩプロモーター活性（Ｂ）が抑制され、かつ上記ＰｔａｃＩプロモーター活性（Ａ）／上記ＰｔａｃＩプロモーター活性（Ｂ）の比の値が、コリネ型細菌（Ｔ）におけるアラビノースを添加した場合のＰｔａｃＩプロモーター活性（Ａ’）／アラビノース未添加の場合のＰｔａｃＩプロモーター活性（Ｂ’）の比の値よりも大きくなる、Condition (III) A coryneform bacterium transformed with a plasmid vector in which the promoter sequences from the 2913th to the 2928th in the promoter vector pGE728-1 shown in SEQ ID NO: 80 are replaced with the above promoter sequence (Y). In S), the PtacI promoter activity (B) when arabinose was added was suppressed with respect to the PtacI promoter activity (A) when arabinose was not added, and the PtacI promoter activity (A) / PtacI promoter activity (B) was suppressed. ) Is larger than the ratio value of PtacI promoter activity (A') when arabinose is added in the coryneform bacterium (T) / PtacI promoter activity (B') when arabinose is not added.
但し、上記コリネ型細菌（Ｔ）は、上記プラスミドベクターｐＧＥ７２８－１のヌクレオチド配列における第２９１３番目から第２９２８番目までのヌクレオチド配列を配列番号１０に記載のヌクレオチド配列と入れ替えたプラスミドベクターで形質転換されたコリネ型細菌であるものとする。However, the coryneform bacterium (T) was transformed with a plasmid vector in which the nucleotide sequence from the 2913th to the 2928th sequence in the nucleotide sequence of the plasmid vector pGE728-1 was replaced with the nucleotide sequence set forth in SEQ ID NO: 10. It is assumed that it is a plasmid-type bacterium.
A method for producing a target substance, which comprises expressing the target gene by exposing the bacterium according to any one of claims 26 to 29 to arabinose or an analog thereof.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2018021453 | 2018-02-08 | ||
| JP2018021453 | 2018-02-08 | ||
| PCT/JP2019/004378 WO2019156152A1 (en) | 2018-02-08 | 2019-02-07 | Nucleic acid including arabinose-dependent gene expression control sequence |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JPWO2019156152A1 JPWO2019156152A1 (en) | 2021-02-12 |
| JPWO2019156152A5 true JPWO2019156152A5 (en) | 2022-02-09 |
| JP7374475B2 JP7374475B2 (en) | 2023-11-07 |
Family
ID=67548309
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2019570794A Active JP7374475B2 (en) | 2018-02-08 | 2019-02-07 | Nucleic acid containing an arabinose-dependent gene expression control sequence |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JP7374475B2 (en) |
| WO (1) | WO2019156152A1 (en) |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008001597A1 (en) * | 2006-06-30 | 2008-01-03 | Ajinomoto Co., Inc. | Method for production of protein |
| RU2433180C2 (en) * | 2008-04-10 | 2011-11-10 | Корея Эдванст Инститьют Оф Сайенс Энд Текнолоджи | Microorganism, which has possibility to produce putrescine in high concentration, method of said microorganism obtaining and method of putrescine production with application of said microorganism |
| MY173771A (en) | 2011-11-11 | 2020-02-20 | Ajinomoto Kk | A method for producing a target substance by fermentation |
-
2019
- 2019-02-07 WO PCT/JP2019/004378 patent/WO2019156152A1/en active Application Filing
- 2019-02-07 JP JP2019570794A patent/JP7374475B2/en active Active
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6805353B2 (en) | A novel adenylosuccinate synthetase and a method for producing purine nucleotides using the synthetase. | |
| Cramer et al. | Phenylalanine ammonia-lyase gene organization and structure | |
| Song et al. | Requirement of a 3′-Terminal Stem-loop inin VitroTranscription by an RNA-dependent RNA Polymerase | |
| Cramer et al. | Identification of RamA, a novel LuxR-type transcriptional regulator of genes involved in acetate metabolism of Corynebacterium glutamicum | |
| JP6652687B2 (en) | Microorganism producing 5'-inosinic acid and method for producing 5'-inosinic acid using the same | |
| CN109022449B (en) | Cucumber CsMLO1 gene and construction method and application of silent expression vector thereof | |
| US20180016609A1 (en) | A Process for the Preparation of Nucleic Acid by Means of 3'-O-Azidomethyl Nucleotide Triphosphate | |
| JP2021019645A (en) | Novel polypeptide and method of producing imp using the same | |
| JP2015536680A (en) | Creation of molecules | |
| CN112266420B (en) | Plant efficient cytosine single-base editor and construction and application thereof | |
| Ray et al. | An internally located RNA hairpin enhances replication of Tomato bushy stunt virus RNAs | |
| AU2019252204A1 (en) | Novel small activating RNA | |
| Miki et al. | Organization of the lexA gene of Escherichia coli and nucleotide sequence of the regulatory region | |
| Hashiro et al. | High copy number mutants derived from Corynebacterium glutamicum cryptic plasmid pAM330 and copy number control | |
| Pátek et al. | Function of Corynebacterium glutamicum promoters in Escherichia coli, Streptomyces lividans, and Bacillus subtilis | |
| KR102617168B1 (en) | Shewanella oneidensis-Drived Protein Expressing Microorganism and Method of L-Amino Acid Production Using the Same | |
| JPWO2019156152A5 (en) | ||
| Benders et al. | Transcriptional analysis of the conserved ftsZ gene cluster in Mycoplasma genitalium and Mycoplasma pneumoniae | |
| JP2022519308A (en) | Mini-circle-producing bacteria engineered to differentially methylate the nucleic acid molecules in them | |
| CN110699487A (en) | Antisense RNA for diagnosing avian leukosis virus tumorigenesis, preparation method, application and primer for constructing over-expression vector | |
| JP3449758B2 (en) | Insertion sequence | |
| O'Connell et al. | Transcriptional organization and regulation of a polycistronic cold shock operon in Sinorhizobium meliloti RM1021 encoding homologs of the Escherichia coli major cold shock gene cspA and ribosomal protein gene rpsU | |
| CN106544313B (en) | A kind of Escherichia coli MG1655 bacterial strain lacking sahn gene and construction method and application | |
| AU2008211580B2 (en) | Use of double stranded RNA hairpin duplexes in gene silencing | |
| Macías et al. | Biology of Trypanosoma cruzi retrotransposons: from an enzymatic to a structural point of view |