WO2001018219A1 - Sequences regulatrices fonctionnant dans des champignons filamenteux - Google Patents
Sequences regulatrices fonctionnant dans des champignons filamenteux Download PDFInfo
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- WO2001018219A1 WO2001018219A1 PCT/JP2000/006104 JP0006104W WO0118219A1 WO 2001018219 A1 WO2001018219 A1 WO 2001018219A1 JP 0006104 W JP0006104 W JP 0006104W WO 0118219 A1 WO0118219 A1 WO 0118219A1
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- nucleotide sequence
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- evening
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01031—Beta-glucuronidase (3.2.1.31)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
Definitions
- the present invention relates to promoters and termines that function in filamentous fungi, expression vectors comprising them, and hosts transformed thereby.
- the filamentous fungus PF102 strain (Mycelia sterilia) produces a PF102 substance which is a 24-membered cyclic depsipeptide with anthelmintic activity (FERM BP-2671). Since this strain does not form sexual and asexual genitalia, it is classified as incomplete sporeless fungus (Japanese Patent Publication No. 3-35796).
- a transformant of the PF102 strain was obtained by introducing a plasmid into which the amylase gene derived from Aspergillus oryzae was linked, together with drug resistance, into the PF102 strain. (W097 / 00944).
- the control DM sequence of the evening amylase gene derived from Aspergillus oryzae shown in WO 97/94444 is a control DM sequence derived from a heterologous strain.
- the genetic characteristics of Mycelia ⁇ ⁇ ⁇ have not yet been fully elucidated, and the conditions to be satisfied by the expression vector system are unknown.
- the expression control of the transformant using the control sequence derived from the heterologous strain is synchronized with the expression of the gene endogenous to the PF122 strain.
- the PF102 strain belongs to the aspergillus-deficient bacterium, it is used in the genus Trichoderma, the genus Fusarium, and the genus Neurospora other than the genus Aspergillus. It is also unclear whether some conventional regulatory DNA sequences are purposefully expressed.
- An object of the present invention is to provide a regulatory sequence that functions in synchrony with the expression of an endogenous gene in a filamentous fungus belonging to the aspergillus incomplete bacterium, particularly Mycelia sterilia.
- Another object of the present invention is to provide an expression vector that highly expresses a target protein in a filamentous fungus belonging to the aspergillus-deficient bacterium, particularly Mycelia sterilia.
- Still another object of the present invention is to provide a transformed filamentous fungus belonging to the incomplete spore-free bacterium, particularly Mycelia sterilia, which highly produces the target protein.
- Another object of the present invention is to provide a method for producing a protein of interest in a filamentous fungus belonging to the incomplete spore-free bacterium, particularly Mycelia sterilia.
- the present inventors have succeeded in isolating and identifying a gene (Abpl gene) expressed at a high rate and a regulatory DNA sequence thereof in the PF102 strain.
- the present inventors also prepared an expression vector for gene expression using the obtained control DNA sequence, and introduced this into a PF102 substance producing bacterium to obtain a transformant. We succeeded in efficiently expressing the target gene linked downstream without impairing its function.
- a promoter according to the present invention is one comprising a nucleotide sequence selected from the group consisting of: and a fragment thereof having promoter activity:
- the evening minerals according to the invention are those comprising a nucleotide sequence selected from the group consisting of: and a fragment thereof having evening mineral activity: (e) nucleotides of SEQ ID NO: 2 Array,
- (f) has at least 70% identity to the nucleotide sequence of SEQ ID NO: 2, and A nucleotide sequence having an evening-mine activity,
- (h) a nucleotide sequence that hybridizes with the nucleotide sequence of SEQ ID NO: 2 under stringent conditions and has an activity of one-minute to one-minute.
- the expression vector according to the present invention comprises one or both of the promoter or a fragment thereof and the evening or mineral or a fragment thereof.
- the transformed host according to the present invention has been transformed by the above expression vector.
- the method for producing a target substance according to the present invention comprises culturing the above-mentioned transformed host, and then collecting the target protein from the culture.
- FIG. 1 shows a restriction map of a 6 kb iidlll fragment containing the gene.
- FIG. 2 shows the structure and restriction map of pABPd.
- the PF 1022 strain was deposited on January 24, 1989 with the Institute of Life Science and Industrial Technology, Institute of Industrial Science and Technology, Ministry of International Trade and Industry (1-1-3 Tsukuba East, Ibaraki, Japan).
- the accession number is FERM BP-2671.
- control sequences that function in a PF1022 producing bacterium, that is, a promoter and a terminator.
- sequence (b) the identity with the nucleotide sequence of SEQ ID NO: 1 can be preferably at least 80%, more preferably at least 90%, most preferably at least 95%.
- sequence (f) the identity with the nucleotide sequence of SEQ ID NO: 2 can preferably be at least 80%, more preferably at least 90%, most preferably at least 95%.
- sequence (c) and sequence (g) the number of modifications is, for example, 1 to several tens be able to.
- the types of the introduced modifications may be the same or different.
- stringent conditions means that the membrane washing operation after hybridization is performed in a low salt concentration solution at a high temperature.
- 5XSSC concentration lxSSC: 15 mM sodium citrate, 150 mM sodium chloride
- 60 ° C for 15 minutes
- 0.5XSSC concentration 60 ° C in 0.1% SDS solution
- Washing conditions for 15 minutes.
- a fragment having promoter activity can be at least 600 base pairs in length, preferably at least 800 base pairs, more preferably at least 1000 base pairs, and most preferably at least 1200 base pairs.
- a fragment having overnight activity can be at least 400 base pairs in length, preferably at least 600 base pairs, more preferably at least 800 base pairs, and most preferably 1000 base pairs.
- sequences (b), (c) and (d), and the fragment having promoter activity are determined by, for example, using an expression vector as described in Example 3. It can be evaluated by preparing, expressing a heterologous gene in a host as described in Example 4, and detecting the production of the heterologous protein. With respect to the sequences (f), (g) and (h), and the fragment having the evening-mine / one-night activity, whether or not “having the evening-mine-one-night activity” is described in Example 3, for example. As described in Example 4, the expression can be evaluated by expressing a heterologous gene in a host and detecting the production of the heterologous protein as described in Example 4.
- the promoter and the terminator according to the present invention can function in a filamentous fungus classified as an incomplete spore-free bacterium, particularly a microorganism belonging to Mycelia, more specifically, a microorganism belonging to Mycelia sterilia.
- Promote and Mineral according to the present invention can function in PF1022 producing bacteria.
- the PF 1022 producing bacteria include filamentous fungi classified as incomplete spore-free bacteria that produce PF 1022 substances.
- the promoter according to the present invention and the minerals can be obtained, for example, as follows.
- the mRNA of the PF102 strain is isolated from the cells at the time of the production of the PF102 substance, and is used as type I to synthesize cDNA. They are selected at random and subjected to nucleotide sequence analysis to isolate a gene expressed at a high frequency, that is, cDNA derived from the Abpl gene.
- Genomic DNA is extracted from the PF102 strain and digested with appropriate restriction enzymes, and then a library consisting of the genomic DNA of the PF102 substance-producing bacterium is extracted using a phage vector or a plasmid vector. Is prepared.
- the full length Abpl gene is cloned using the translation region encoding the Abpl gene as a probe.
- the promoter of the present gene and a portion of the motif can be determined to obtain the promoter and the motif. it can.
- an expression vector comprising a control sequence that functions in a PF102 substance producing bacterium.
- the procedures and methods for constructing the expression vector according to the present invention may be those commonly used in the field of genetic engineering.
- the copy number of the gene present in the host cell may be one copy or plural.
- the expression vector according to the present invention comprises, in a first aspect thereof, the promoter according to the present invention and / or the mineral and / or optionally a genetic marker and / or other regulatory sequences. It comprises. Accordingly, an expression vector comprising one or both of the promoters of the present invention Is included in the scope of the present invention.
- the evening and the evening may be other than the evening and the evening according to the present invention.
- the promoter may be other than the promoter according to the present invention.
- the gene marker can be introduced, for example, by introducing an appropriate restriction enzyme cleavage site into the control sequence of the present invention by PCR and inserting it into a plasmid vector, followed by drug resistance gene and / or auxotrophic complement gene. This can be done by linking selection genes such as
- the gene can be appropriately selected depending on the method for selecting a transformant.
- a gene encoding drug resistance or a gene complementing auxotrophy can be used.
- drug resistance genes include genes for drugs such as destomycin, benomyl, oligomycin, hygromycin, G418, bleomycin, vialaphos, plasticidin S, phleomycin, phosphinothricin, ambicilin, and kanamycin.
- Genes that complement auxotrophy include ⁇ , rGs argBs trpC, niaD ⁇ TRPU LEU2, URA3, and other genes.
- the expression vector according to the present invention in its second embodiment, may further comprise a nucleotide sequence encoding a protein of interest operably linked to its control sequence.
- Linkage to a control sequence can be performed, for example, by inserting a translation region of a gene encoding a target protein (target gene) downstream of the promoter in a forward direction according to a conventional method.
- the target gene can be linked to a foreign gene encoding a translation region of another protein and expressed as a fusion protein.
- target gene means any gene to be expressed, and may be a heterologous gene or a homologous gene.
- the target gene can be, for example, a gene selected from a group of genes related to PF102 substance production.
- a host transformed with the expression vector can be used as a host for genetic recombination
- the microorganism is a filamentous fungus, preferably a filamentous fungus classified as an asporogenous incomplete bacterium, more preferably a microorganism belonging to Mycelia, and most preferably a microorganism belonging to Mycelia sterilia.
- the host that can be used in the present invention also includes a PF102 substance producing bacterium, preferably a filamentous fungus producing a PF102 substance, and more preferably a PF102 substance producing a PF102 substance. 2 strains (FERM BP-2671).
- Introduction of a recombinant vector for gene expression into a host can be performed according to a conventional method.
- the introduction method include an electrolysis method, a polyethylene glycol method, an agrobacterium method, a lithium method and a calcium chloride method, and an efficient method for a host cell is selected.
- the polyethylene glycol method is preferred.
- a method for producing a target protein comprising a step of culturing the transformant.
- Culturing of the transformant can be performed by appropriately selecting a medium, culture conditions, and the like according to a conventional method.
- a medium conventional components, for example, as a carbon source, glucose, sucrose, cellulose, starch syrup, dextrin, starch, glycerol, molasses, animal and vegetable oils can be used.
- Nitrogen sources include soybean flour, wheat germ, pharmamedia, corn 'stip' liquor, cottonseed meal, bouillon, leptone, polypeptone, malt extract, yeast extract, ammonium sulfate, sodium nitrate, urea. Etc. can be used.
- sodium, potassium, calcium, magnesium, cobalt, chlorine, phosphoric acid, sulfuric acid, and other inorganic salts capable of forming an ion such as potassium chloride, calcium carbonate, dipotassium hydrogen phosphate
- magnesium sulfate, potassium phosphate, zinc sulfate, manganese sulfate, and copper sulfate is also effective to add magnesium sulfate, potassium phosphate, zinc sulfate, manganese sulfate, and copper sulfate.
- trace nutrients such as various amino acids such as thiamine (such as thiamine hydrochloride), amino acids such as glutamic acid (such as sodium glutamate) and nucleotides such as nucleotides such as thiamine (such as thiamine hydrochloride).
- Selective drugs such as antibiotics can also be added.
- the culture method of the liquid medium can be performed by aerobic culture, shaking culture, aeration stirring culture, or submerged culture.
- the pH of the medium is, for example, about pH 6 to pH 8.
- the culturing can be performed under ordinary conditions, for example, a temperature of 14 ° C to 40 °, preferably 26 ° C to 37 ° C, and the culturing can be performed under the conditions of about 2 to 25 days.
- a protein which is a gene expression product of interest can be obtained from a culture of the transformed cells.
- the target protein can be extracted (trituration, pressure crushing, etc.) from the culture, recovered (filtration, centrifugation, etc.), and purified (salting out, solvent precipitation, etc.).
- a protease inhibitor such as phenylmethylsulfonyl fluoride (PMSF), benzamidine or leptin can be added as necessary.
- Example 1 Search for highly expressed genes by random sequence of cDNA
- cDNA derived from bacteria producing PF1022 substance is randomly cloned, their DNA sequences are compared, and large quantities are expressed. Genes were isolated and identified.
- Production medium for PF 1022 strain (FERM BP-2671) (glucose 2.0%, starch 5.0%, wheat germ 0.8%, soybean meal 1.3%, meat extract 0.38%, sodium chloride 0.3%). 13% and calcium carbonate 0.15%; pH before sterilization is 7.0; see W097 / 00944, Example 4) at 26 ° C for 4 days and centrifuged (3000 rpm, 10 minutes). The body was recovered. This was washed with purified water, frozen at -80 ° C, and then pulverized with a blender (Nippon Seiki AM-3) in the presence of liquid nitrogen.
- a blender Nippon Seiki AM-3
- RNA fraction was dissolved in TE (10 mM Tris-HCl (pH 8.0), ImM EDTA) so as to have a nucleic acid concentration of lmg / ml, and precipitated with 2.5 M lithium chloride (5 ° C, 2 hours). This was collected by centrifugation, washed with 70 ethanol, and redissolved in TE, which was used as a total RNA fraction.
- TE Tris-HCl (pH 8.0), ImM EDTA
- MRNA was purified from all RM fractions using mRNA Purification Kit (Amersham Pharmacia Biotech). Furthermore, cDNA was synthesized using this mRNA as a type II using a Timesaver cDNA synthesis kit (manufactured by Amersham Pharmacia Biotech).
- the cDNA prepared as in Example 1 (1) was cut and ligated to PUC18 treated with alkaline phosphatase using a DNA ligase kit Ver.2 (Takara Shuzo). This was introduced into Escherichia coli JM109 strain, and various transformed colonies were cultured in an LB medium (1% polypeptone, 0.5% yeast extract, 1% sodium chloride) containing ambicillin. Purification of the plasmid from these transformants was performed using a flexibrop kit (manufactured by Amersham Pharmacia Biotech).
- the 40 kinds of plasmids prepared as described above were subjected to ALF DNA Sequencer II (Amersham Pharmacia Biotech) to decode the DNA sequence of the inserted fragment.
- a long ranger (manufactured by FMC) was used for the sequencing gel, and an automatic sequencing kit (manufactured by Amersham Pharmacia Biotech) was used for the sequencing reaction.
- the cloned gene was named and the promoter and terminator of this gene were cloned from genomic DNA.
- Genomic DM of the PF 1022 strain was isolated according to the method described in (H, Horiuchi et. Al., J. BacterioL, 170, 272-278, (1988)). Specifically, first, a PF 1022 substance producing bacterium (FERM BP-2671) was added to a seed medium (soluble starch 2.0%, glucose 1.0%, polypeptone 0.5%, wheat germ 0.6%, yeast extract 0%). 3%, soybean meal 0.2% and calcium carbonate 0.2%; kill The cells were cultured at pH 7.0; W097 / 00944, Example 1) for 2 days, and the cells were collected by centrifugation (3,500 rpm, 10 minutes).
- the obtained cells were freeze-dried, suspended in TE, treated in a 3 SDS solution at 60 ° C. for 30 minutes, and the cell residues were removed by TE-saturated phenol extraction.
- the extract was precipitated with ethanol, treated with ribonuclease A (manufactured by Sigma) and proteinase K (manufactured by Wako Pure Chemical Industries), and the nucleic acid was precipitated with 12% polyethylene glycol 6000. This was subjected to TE-saturated phenol extraction and ethanol precipitation, and the precipitate was dissolved in TE to obtain genomic DNA.
- the genomic DNA derived from the PF102 substance producing bacterium prepared as in Example 1 (3) was partially digested with Sau3AI. This was ligated to the I-arm of a phage vector, Human EMBL3 Cloning Kit (Stratagene) using T4 ligase (Ligation kit Ver.2, Takara Shuzo). This was dissolved in TE after ethanol precipitation. The whole amount of the ligation mixture was used to infect Escherichia coli strain LE392 using a Gigapack II Plus packaging kit (Stratagene) to form phage plaques. Genes were cloned using 1.3 ⁇ 10 ′ (2.6 ⁇ l (TPFU / ml)) phage libraries obtained by this method.
- the genomic DNA prepared as in (3) was subjected to PCR in accordance with Lego-I PCR kit (manufactured by Sadei Technology Co., Ltd.) using the 8-73U and 8-73R synthetic primers.
- amplification was performed by repeating a step of 94 ° C for 30 seconds, 50 ° C for 30 seconds and 72 ° C for 90 seconds 25 times.
- the DNA sequences of 8-73U and 8-73R are shown below.
- PCR product thus obtained was labeled using an ECL direct system (Amersham Pharmacia Biotech). Phage plaques prepared as in Example 1 (4) were transferred to Hybond N + nylon transfer membrane (manufactured by Amersham-Pharmacia Biotech), denatured with alkali, and 5 times concentrated SSC
- pRQHin / 119 was digested with l and l, ligated to pUC18 digested with the same restriction enzymes, and used as a template for DNA sequence analysis.
- DM sequence analysis was performed in the same manner as in Example 1 (2). Next, by comparing with the nucleotide sequence of the cDNA obtained in Example 1 (2), the promoter of the Abpl gene and the evening region were determined. It is shown in SEQ ID NO: 2.
- the promoter evening region and the evening mineral region were amplified using the PCH method.
- Amplification of promoters is performed using ABP-Neco and ABP-Nbam, while amplification of evening and night is performed using primers ABP-Cbam and ABP-Cxba, and PCR Supermix High Fidelity (manufactured by Lifetech Oriental).
- PCR was performed by As the reaction conditions, amplification was performed by repeating a step of 94 ° C for 30 seconds, 50 ° C for 30 seconds, and 72 ° C for 90 seconds 25 times.
- the DM sequences of ABP-Neco, ABP-Nbam, ABP-Cbam and ABP-Cxba are shown below.
- ABP-Neco GGGGAATTCGTGGGTGGTGATATCATGGC (SEQ ID NO: 3)
- ABP-Nbajn GGGGGATCCTTGATGGGTTTTGGG (SEQ ID NO: 4)
- ABP-Cbam GGGGGATCCTAAACTCCCATCTATAGC (SEQ ID NO: 5)
- ABP-Cxba GGGTCTAGACGACTCATTGCAGTGAGTGG (SEQ ID NO: 6)
- PCR product was purified on a Microsvin S-400 column (Amersham Pharmacia Biotech), and after ethanol precipitation, the promoters were EcoRI and BamHI, and the -Mine overnight was digested with BamHI and Xal and ligated sequentially to pBluescriptll KS + digested with the same enzymes. This was digested with ⁇ I, and a pMKDOl (W098 / 03667) -derived destomycin resistance cassette was inserted to construct pABPd (Fig. 2).
- the translation region of the? -Glucuronidase ( ⁇ ) gene used as the reporter gene was PLC-GUS (K. Yanai, et. Al., Biosci. Biotech. Biochem., 60, 472-475,
- the transformant thus obtained was subjected to liquid culture using the production medium of Example 1 (1), and the cells were recovered by centrifugation.
- the cells of the obtained cells were disrupted using Mini Bead Douichi (manufactured by BioSpec Products). This was centrifuged to remove cell debris, and the GUS activity of the supernatant was measured. The activity was measured according to the method described in (K. Yanai, et. Al., Biosci. Biotech. Biochem., 60, 472-475, (1996)).
- Table 1 GUS activity of transformant Expression vector GUS activity (A405 / ⁇ g protein) Transformant pABPd_G
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Description
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Priority Applications (9)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE60032162T DE60032162T2 (de) | 1999-09-07 | 2000-09-07 | Regulatorische sequenzen von funktioneller bedeutung in filamentösen pilzen |
US10/070,386 US6913905B1 (en) | 1999-09-07 | 2000-09-07 | Regulatory sequences functioning in filamentous fungi |
NZ517463A NZ517463A (en) | 1999-09-07 | 2000-09-07 | Promoter and terminator sequences functioning in filamentous fungi |
JP2001521755A JP4672222B2 (ja) | 1999-09-07 | 2000-09-07 | 糸状菌において機能する制御配列および発現系 |
KR1020027002705A KR20020057951A (ko) | 1999-09-07 | 2000-09-07 | 사상균에서 기능하는 제어서열 및 발현계 |
CA002384000A CA2384000A1 (en) | 1999-09-07 | 2000-09-07 | Regulatory sequences and expression systems functioning in filamentous fungi |
AU68742/00A AU782525B2 (en) | 1999-09-07 | 2000-09-07 | Regulatory sequences functioning in filamentous fungi |
EP00957010A EP1221489B1 (en) | 1999-09-07 | 2000-09-07 | Regulatory sequences functioning in filamentous fungi |
NO20020941A NO331102B1 (no) | 1999-09-07 | 2002-02-27 | Promotor, ekspresjonsvektor og vert, og fremgangsmate for a produsere et aktuelt protein |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP25285199 | 1999-09-07 | ||
JP11/252851 | 1999-09-07 |
Publications (1)
Publication Number | Publication Date |
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WO2001018219A1 true WO2001018219A1 (fr) | 2001-03-15 |
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ID=17243068
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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PCT/JP2000/006104 WO2001018219A1 (fr) | 1999-09-07 | 2000-09-07 | Sequences regulatrices fonctionnant dans des champignons filamenteux |
Country Status (12)
Country | Link |
---|---|
US (1) | US6913905B1 (ja) |
EP (1) | EP1221489B1 (ja) |
JP (1) | JP4672222B2 (ja) |
KR (1) | KR20020057951A (ja) |
CN (1) | CN100374571C (ja) |
AT (1) | ATE346943T1 (ja) |
AU (1) | AU782525B2 (ja) |
CA (1) | CA2384000A1 (ja) |
DE (1) | DE60032162T2 (ja) |
NO (1) | NO331102B1 (ja) |
NZ (1) | NZ517463A (ja) |
WO (1) | WO2001018219A1 (ja) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001081563A1 (fr) * | 2000-04-26 | 2001-11-01 | Meiji Seika Kaisha, Ltd. | Nouvelle (r)-2-hydroxy-3-phenylpropionate (d-phenyl-lactate) deshydrogenase et gene codant pour celle-ci |
US7659065B2 (en) | 2003-02-12 | 2010-02-09 | Ramot At Tel Aviv University Ltd. | Transgenic fungi expressing Bcl-2 and methods of using Bcl-2 or portions thereof for improving biomass production, survival, longevity and stress resistance of fungi |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN102559678B (zh) * | 2011-12-26 | 2013-04-17 | 浙江大学 | 一种丝状真菌启动子和包含该启动子的质粒 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0382173A2 (en) * | 1989-02-07 | 1990-08-16 | Meiji Seika Kaisha Ltd. | PF 1022 substance, method of producing same and anthelmintic composition containing same |
JPH07123987A (ja) * | 1993-11-08 | 1995-05-16 | Amano Pharmaceut Co Ltd | 糸状菌および酵母で使用可能なポリペプチド分泌発現用プラスミドおよびそれを用いたポリペプチドの製造法 |
JPH11276170A (ja) * | 1998-03-31 | 1999-10-12 | Amano Pharmaceut Co Ltd | アスペルギルス属由来の新規なプロモーター |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0780468A4 (en) | 1995-06-22 | 2000-07-19 | Meiji Seika Kaisha | TRANSFORMANT WHICH PRODUCES THE SUBSTANCE PF1022 AND METHOD FOR TRANSFORMING MICROORGANISMS IN THE CLASS OF THE HYPOMYCETES |
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2000
- 2000-09-07 KR KR1020027002705A patent/KR20020057951A/ko not_active Application Discontinuation
- 2000-09-07 AT AT00957010T patent/ATE346943T1/de not_active IP Right Cessation
- 2000-09-07 WO PCT/JP2000/006104 patent/WO2001018219A1/ja active IP Right Grant
- 2000-09-07 EP EP00957010A patent/EP1221489B1/en not_active Expired - Lifetime
- 2000-09-07 AU AU68742/00A patent/AU782525B2/en not_active Ceased
- 2000-09-07 CN CNB008153043A patent/CN100374571C/zh not_active Expired - Fee Related
- 2000-09-07 CA CA002384000A patent/CA2384000A1/en not_active Abandoned
- 2000-09-07 DE DE60032162T patent/DE60032162T2/de not_active Expired - Lifetime
- 2000-09-07 JP JP2001521755A patent/JP4672222B2/ja not_active Expired - Fee Related
- 2000-09-07 US US10/070,386 patent/US6913905B1/en not_active Expired - Fee Related
- 2000-09-07 NZ NZ517463A patent/NZ517463A/en not_active IP Right Cessation
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2002
- 2002-02-27 NO NO20020941A patent/NO331102B1/no not_active IP Right Cessation
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0382173A2 (en) * | 1989-02-07 | 1990-08-16 | Meiji Seika Kaisha Ltd. | PF 1022 substance, method of producing same and anthelmintic composition containing same |
JPH07123987A (ja) * | 1993-11-08 | 1995-05-16 | Amano Pharmaceut Co Ltd | 糸状菌および酵母で使用可能なポリペプチド分泌発現用プラスミドおよびそれを用いたポリペプチドの製造法 |
JPH11276170A (ja) * | 1998-03-31 | 1999-10-12 | Amano Pharmaceut Co Ltd | アスペルギルス属由来の新規なプロモーター |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001081563A1 (fr) * | 2000-04-26 | 2001-11-01 | Meiji Seika Kaisha, Ltd. | Nouvelle (r)-2-hydroxy-3-phenylpropionate (d-phenyl-lactate) deshydrogenase et gene codant pour celle-ci |
US6916641B2 (en) | 2000-04-26 | 2005-07-12 | Meiji Seika Kaisha, Ltd. | (R)-2-hydroxy-3-phenylpropionate (d-phenyllactate) dehydrogenase and gene encoding the same |
KR100808307B1 (ko) * | 2000-04-26 | 2008-02-27 | 메이지 세이카 가부시키가이샤 | 신규한(알)-2-히드록시-3-페닐프로피온산(디-페닐유산)탈수소효소 및 이 것을 코드하는 유전자 |
JP4679785B2 (ja) * | 2000-04-26 | 2011-04-27 | 明治製菓株式会社 | 新規な(r)−2−ヒドロキシ−3−フェニルプロピオン酸(d−フェニル乳酸)脱水素酵素およびそれをコードする遺伝子 |
US7659065B2 (en) | 2003-02-12 | 2010-02-09 | Ramot At Tel Aviv University Ltd. | Transgenic fungi expressing Bcl-2 and methods of using Bcl-2 or portions thereof for improving biomass production, survival, longevity and stress resistance of fungi |
Also Published As
Publication number | Publication date |
---|---|
AU782525B2 (en) | 2005-08-04 |
JP4672222B2 (ja) | 2011-04-20 |
EP1221489A1 (en) | 2002-07-10 |
EP1221489B1 (en) | 2006-11-29 |
CN1387573A (zh) | 2002-12-25 |
EP1221489A4 (en) | 2003-05-28 |
DE60032162D1 (de) | 2007-01-11 |
AU6874200A (en) | 2001-04-10 |
NO20020941D0 (no) | 2002-02-27 |
ATE346943T1 (de) | 2006-12-15 |
US6913905B1 (en) | 2005-07-05 |
NO20020941L (no) | 2002-05-06 |
DE60032162T2 (de) | 2007-10-25 |
NZ517463A (en) | 2003-10-31 |
NO331102B1 (no) | 2011-10-03 |
CA2384000A1 (en) | 2001-03-15 |
CN100374571C (zh) | 2008-03-12 |
KR20020057951A (ko) | 2002-07-12 |
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