| (11) | EP 3 059 317 B1 |
(12) | EUROPEAN PATENT SPECIFICATION |
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(54) | VECTOR COMPRISING SPECIFIC PROMOTER AND GENE ENCODING SPECIFIC PROTEIN, TRANSGENIC PLANT INTO WHICH THE VECTOR HAS BEEN INTRODUCED, AND METHOD FOR IMPROVING POLYISOPRENOID PRODUCTION BY INTRODUCING THE VECTOR INTO PLANT VEKTOR MIT SPEZIFISCHEM PROMOTOR UND GENCODIERENDEM SPEZIFISCHEN PROTEIN, TRANSGENE PFLANZE MIT DARIN EINGEFÜHRTEM VEKTOR UND VERFAHREN ZUR VERBESSERUNG DER POLYISOPRENOIDPRODUKTION DURCH EINFÜHRUNG DES VEKTORS IN DIE PFLANZE VECTEUR COMPRENANT UN PROMOTEUR SPÉCIFIQUE ET GÈNE CODANT POUR UNE PROTÉINE SPÉCIFIQUE, PLANTE TRANSGÉNIQUE DANS LAQUELLES LE VECTEUR A ÉTÉ INTRODUIT ET PROCÉDÉ PERMETTANT D'AMÉLIORER LA PRODUCTION DE POLYISOPRÉNOÏDE PAR INTRODUCTION DU VECTEUR DANS UNE PLANTE |
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Note: Within nine months from the publication of the mention of the grant of the European patent, any person may give notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall be filed in a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). |
TECHNICAL FIELD
BACKGROUND ART
SUMMARY OF INVENTION
TECHNICAL PROBLEM
SOLUTION TO PROBLEM
[A1] a DNA comprising the base sequence of base numbers 1 to 1680 represented by SEQ ID NO: 1;
[A2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1680 represented by SEQ ID NO: 1 under stringent conditions, and having a promoter activity for laticifer-specific gene expression; and
[A3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1680 represented by SEQ ID NO: 1, and having a promoter activity for laticifer-specific gene expression.
[B1] a DNA comprising the base sequence of base numbers 1 to 1029 represented by SEQ ID NO: 2;
[B2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1029 represented by SEQ ID NO: 2 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates or a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates; and
[B3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1029 represented by SEQ ID NO: 2, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates or a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates.
[C1] a DNA comprising the base sequence of base numbers 1 to 1113 represented by SEQ ID NO: 4;
[C2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1113 represented by SEQ ID NO: 4 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates, a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates, or a reaction using isopentenyl diphosphate and farnesyl diphosphate as substrates; and
[C3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1113 represented by SEQ ID NO: 4, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates, a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates, or a reaction using isopentenyl diphosphate and farnesyl diphosphate as substrates.
[D1] a DNA comprising the base sequence of base numbers 1 to 1728 represented by SEQ ID NO: 6;
[D2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1728 represented by SEQ ID NO: 6 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1728 represented by SEQ ID NO: 6, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D4] a DNA comprising the base sequence of base numbers 1 to 1761 represented by SEQ ID NO: 7;
[D5] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1761 represented by SEQ ID NO: 7 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D6] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1761 represented by SEQ ID NO: 7, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D7] a DNA comprising the base sequence of base numbers 1 to 1821 represented by SEQ ID NO: 8;
[D8] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1821 represented by SEQ ID NO: 8 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D9] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1821 represented by SEQ ID NO: 8, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D10] a DNA comprising the base sequence of base numbers 1 to 1581 represented by SEQ ID NO: 9;
[D11] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1581 represented by SEQ ID NO: 9 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA; and
[D12] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1581 represented by SEQ ID NO: 9, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA.
[E1] a DNA comprising the base sequence of base numbers 1 to 705 represented by SEQ ID NO: 14;
[E2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 705 represented by SEQ ID NO: 14 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of isomerization of isopentenyl diphosphate or dimethylallyl diphosphate; and
[E3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 705 represented by SEQ ID NO: 14, and encoding a protein having an enzyme activity that catalyzes a reaction of isomerization of isopentenyl diphosphate or dimethylallyl diphosphate.
[F1] a DNA comprising the base sequence of base numbers 1 to 873 represented by SEQ ID NO: 16;
[F2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 873 represented by SEQ ID NO: 16 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[F3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 873 represented by SEQ ID NO: 16, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[F4] a DNA comprising the base sequence of base numbers 1 to 855 represented by SEQ ID NO: 66;
[F5] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 855 represented by SEQ ID NO: 66 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds; and
[F6] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 855 represented by SEQ ID NO: 66, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds.
[G1] a DNA comprising the base sequence of base numbers 1 to 615 represented by SEQ ID NO: 18;
[G2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 615 represented by SEQ ID NO: 18 under stringent conditions, and encoding a rubber particle-associated protein which is associated with rubber particles in latex; and
[G3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 615 represented by SEQ ID NO: 18, and encoding a rubber particle-associated protein which is associated with rubber particles in latex.
[H1] a DNA comprising the base sequence of base numbers 1 to 417 represented by SEQ ID NO: 60;
[H2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 417 represented by SEQ ID NO: 60 under stringent conditions, and encoding a rubber particle-associated protein which is associated with rubber particles in latex; and
[H3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 417 represented by SEQ ID NO: 60, and encoding a rubber particle-associated protein which is associated with rubber particles in latex.
ADVANTAGEOUS EFFECTS OF INVENTION
BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 is a schematic diagram showing a part of the polyisoprenoid biosynthesis pathway;
FIG. 2 is a schematic diagram showing a part of the polyisoprenoid biosynthesis pathway;
FIG. 3 is a schematic diagram showing a part of the mevalonic acid pathway which is upstream of a polyisoprenoid synthesis pathway;
FIG. 4 is a schematic diagram showing a part of the polyisoprenoid biosynthesis pathway;
FIG. 5 is a schematic diagram showing a part of the polyisoprenoid biosynthesis pathway;
FIG. 6 is a schematic diagram showing a part of the polyisoprenoid biosynthesis pathway;
FIG. 7 is a schematic diagram showing a T-DNA region of an expression vector introduced into Agrobacterium used in Example 1;
FIG. 8 is a schematic diagram showing a T-DNA region of an expression vector introduced into Agrobacterium used in Example 2;
FIG. 9 is a schematic diagram showing a T-DNA region of an expression vector introduced into Agrobacterium used in Example 3, 8, 9, or 10;
FIG. 10 is a schematic diagram showing a T-DNA region of an expression vector introduced into Agrobacterium used in Example 4;
FIG. 11 is a schematic diagram showing a T-DNA region of an expression vector introduced into Agrobacterium used in Example 5 or 11;
FIG. 12 is a schematic diagram showing a T-DNA region of an expression vector introduced into Agrobacterium used in
Example 6; and
DESCRIPTION OF EMBODIMENTS
(Vector)
(Hevein 2.1 Encoding Gene Promoter)
[A1] a DNA comprising the base sequence of base numbers 1 to 1680 represented by SEQ ID NO: 1;
[A2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1680 represented by SEQ ID NO: 1 under stringent conditions, and having a promoter activity for laticifer-specific gene expression; and
[A3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1680 represented by SEQ ID NO: 1, and having a promoter activity for laticifer-specific gene expression.
(Protein Involved in Polyisoprenoid Biosynthesis)
(Gene)
[B1] a DNA comprising the base sequence of base numbers 1 to 1029 represented by SEQ ID NO: 2;
[B2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1029 represented by SEQ ID NO: 2 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates or a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates; and [B3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1029 represented by SEQ ID NO: 2, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates or a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates.
[C1] a DNA comprising the base sequence of base numbers 1 to 1113 represented by SEQ ID NO: 4;
[C2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1113 represented by SEQ ID NO: 4 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates, a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates, or a reaction using isopentenyl diphosphate and farnesyl diphosphate as substrates; and
[C3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1113 represented by SEQ ID NO: 4, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates, a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates, or a reaction using isopentenyl diphosphate and farnesyl diphosphate as substrates.
[D1] a DNA comprising the base sequence of base numbers 1 to 1728 represented by SEQ ID NO: 6;
[D2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1728 represented by SEQ ID NO: 6 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1728 represented by SEQ ID NO: 6, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D4] a DNA comprising the base sequence of base numbers 1 to 1761 represented by SEQ ID NO: 7;
[D5] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1761 represented by SEQ ID NO: 7 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D6] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1761 represented by SEQ ID NO: 7, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D7] a DNA comprising the base sequence of base numbers 1 to 1821 represented by SEQ ID NO: 8;
[D8] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1821 represented by SEQ ID NO: 8 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D9] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1821 represented by SEQ ID NO: 8, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D10] a DNA comprising the base sequence of base numbers 1 to 1581 represented by SEQ ID NO: 9;
[D11] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1581 represented by SEQ ID NO: 9 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA; and
[D12] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1581 represented by SEQ ID NO: 9, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA.
[E1] a DNA comprising the base sequence of base numbers 1 to 705 represented by SEQ ID NO: 14;
[E2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 705 represented by SEQ ID NO: 14 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of isomerization of isopentenyl diphosphate or dimethylallyl diphosphate; and
[E3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 705 represented by SEQ ID NO: 14, and encoding a protein having an enzyme activity that catalyzes a reaction of isomerization of isopentenyl diphosphate or dimethylallyl diphosphate.
[F1] a DNA comprising the base sequence of base numbers 1 to 873 represented by SEQ ID NO: 16;
[F2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 873 represented by SEQ ID NO: 16 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[F3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 873 represented by SEQ ID NO: 16, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[F4] a DNA comprising the base sequence of base numbers 1 to 855 represented by SEQ ID NO: 66;
[F5] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 855 represented by SEQ ID NO: 66 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds; and
[F6] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 855 represented by SEQ ID NO: 66, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds.
[G1] a DNA comprising the base sequence of base numbers 1 to 615 represented by SEQ ID NO: 18;
[G2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 615 represented by SEQ ID NO: 18 under stringent conditions, and encoding a rubber particle-associated protein which is associated with rubber particles in latex; and
[G3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 615 represented by SEQ ID NO: 18, and encoding a rubber particle-associated protein which is associated with rubber particles in latex.
[H1] a DNA comprising the base sequence of base numbers 1 to 417 represented by SEQ ID NO: 60;
[H2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 417 represented by SEQ ID NO: 60 under stringent conditions, and encoding a rubber particle-associated protein which is associated with rubber particles in latex; and
[H3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 417 represented by SEQ ID NO: 60, and encoding a rubber particle-associated protein which is associated with rubber particles in latex.
(Protein)
[d1] a protein comprising the amino acid sequence of amino acid numbers 1 to 575 represented by SEQ ID NO: 10;
[d2] a protein comprising the amino acid sequence of amino acid numbers 1 to 586 represented by SEQ ID NO: 11;
[d3] a protein comprising the amino acid sequence of amino acid numbers 1 to 606 represented by SEQ ID NO: 12; and
[d4] a protein comprising the amino acid sequence of amino acid numbers 1 to 526 represented by SEQ ID NO: 13.
[f1] a protein comprising the amino acid sequence of amino acid numbers 1 to 290 represented by SEQ ID NO: 17; and
[f4] a protein comprising the amino acid sequence of amino acid numbers 1 to 284 represented by SEQ ID NO: 67.
[b2] a protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 342 represented by SEQ ID NO: 3, and having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates or a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates;
[c2] a protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 370 represented by SEQ ID NO: 5, and having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates, a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates, or a reaction using isopentenyl diphosphate and farnesyl diphosphate as substrates;
[d5] a protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 575 represented by SEQ ID NO: 10, and having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[d6] a protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 586 represented by SEQ ID NO: 11, and having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[d7] a protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 606 represented by SEQ ID NO: 12, and having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[d8] a protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 526 represented by SEQ ID NO: 13, and having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[e2] a protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 234 represented by SEQ ID NO: 15, and having an enzyme activity that catalyzes a reaction of isomerization of isopentenyl diphosphate or dimethylallyl diphosphate;
[f2] a protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 290 represented by SEQ ID NO: 17, and having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[f5] a protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 284 represented by SEQ ID NO: 67, and having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[g2] a rubber particle-associated protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 204 represented by SEQ ID NO: 19, and being associated with rubber particles in latex; and
[h2] a rubber particle-associated protein comprising an amino acid sequence containing one or more amino acid substitutions, deletions, insertions, and/or additions relative to the amino acid sequence of amino acid numbers 1 to 138 represented by SEQ ID NO: 61, and being associated with rubber particles in latex.
[b3] a protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 342 represented by SEQ ID NO: 3, and having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates or a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates;
[c3] a protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 370 represented by SEQ ID NO: 5, and having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates, a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates, or a reaction using isopentenyl diphosphate and farnesyl diphosphate as substrates;
[d9] a protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 575 represented by SEQ ID NO: 10, and having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[d10] a protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 586 represented by SEQ ID NO: 11, and having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[d11] a protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 606 represented by SEQ ID NO: 12, and having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[d12] a protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 526 represented by SEQ ID NO: 13, and having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[e3] a protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 234 represented by SEQ ID NO: 15, and having an enzyme activity that catalyzes a reaction of isomerization of isopentenyl diphosphate or dimethylallyl diphosphate;
[f3] a protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 290 represented by SEQ ID NO: 17, and having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[f6] a protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 284 represented by SEQ ID NO: 67, and having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[g3] a rubber particle-associated protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 204 represented by SEQ ID NO: 19, and being associated with rubber particles in latex; and
[h3] a rubber particle-associated protein comprising an amino acid sequence having 80% or more sequence identity with the amino acid sequence of amino acid numbers 1 to 138 represented by SEQ ID NO: 61, and being associated with rubber particles in latex.
(Transformant)
(Induction Step)
(Infection Step)
(Coculture Step)
(Selective Culture Step)
(Regeneration-inducing Step)
(Elongation Step)
(Rooting Step)
EXAMPLES
(Preparation of Base Sequence of Promoter of Gene Encoding HEV2.1)
(Primer 1: SEQ ID NO: 20)
5'-CTCAAGGCTACCTTATTGGG-3'
(Primer 2: SEQ ID NO: 21)
5'-CTCAGCAATTGCAACACCTG-3'
(Amplification of Promoter Sequence for Introduction into Vector)
(Primer 3: SEQ ID NO: 22)
5'-GGTACCGCTACCTTATTGGGAACTACC-3'
(Primer 4: SEQ ID NO: 23)
5'-AGATCTAACTCTTCCCATTTCTTCCC-3'
(Identification of Amino Acid Sequences and Base Sequences of Target Proteins of Enzymes Involved in Polyisoprenoid Biosynthesis Pathway in Para Rubber Tree)
(Primer 5: SEQ ID NO: 24)
5'-GAATCCATGGCGGATCTGAAG-3'
(Primer 6: SEQ ID NO: 25)
5'-GTCCATGTATCTGGATACCC-3'
(Primer 7: SEQ ID NO: 26)
5'-CAAGATGAGTTCAGTGAATTTGGG-3'
(Primer 8: SEQ ID NO: 27)
5'-TGCATTAGTTTTGCCTGTGAGC-3'
(Primer 9: SEQ ID NO: 28)
5'-ATTTTTACATGGACACCACCG-3'
(Primer 10: SEQ ID NO: 29)
5'-ACCAGATTCCCACTAAGATGC-3'
(Primer 11: SEQ ID NO: 30)
5'-TCCATATATGGACGAGGTTCG-3'
(Primer 12: SEQ ID NO: 31)
5'-GCAGCTGTGTTACCCTTCAG-3'
(Primer 13: SEQ ID NO: 32)
5'-CAGTCGCTCCAAAATGGATGTGC-3'
(Primer 14: SEQ ID NO: 33)
5'-GATTTTCTTAGGAAGAAGGCTTGG-3'
(Primer 15: SEQ ID NO: 34)
5'-CTAGCTGGTCTATAATGGATGCC-3'
(Primer 16: SEQ ID NO: 35)
5'-GAATCAATTTACCTCAATAGAAGGC-3'
(Primer 17: SEQ ID NO: 36)
5'-TTCCACCATGGGTGAGGCTCC-3'
(Primer 18: SEQ ID NO: 37)
5'-TCTCAACTCAACTTGTGAATCG-3'
(Primer 19: SEQ ID NO: 38)
5'-ATGGAATTATACAACGGTGAGAGG-3'
(Primer 20: SEQ ID NO: 39)
5'-TTTTAAGTATTCCTTATGTTTCTCC-3'
(Primer 45: SEQ ID NO: 68)
5'-ATGGAATTATACAACGGTGAGAGG-3'
(Primer 46: SEQ ID NO: 69)
5'-TTTTAAGTATTCCTTATGTTTCTCC-3'
(Primer 21: SEQ ID NO: 40)
5'-TATGGCTGAAGAGGTGGAGG-3'
(Primer 22: SEQ ID NO: 41)
5'-TGATGCCTCATCTCCAAACACC-3'
(Primer 41: SEQ ID NO: 62)
5'-ATGGCTGAAGACGAAGACAACC-3'
(Primer 42: SEQ ID NO: 63)
5'-ATTCTCTCCATAAAACACCTTAGC-3'
(Amplification of Base Sequences of Genes Encoding Target Proteins for Introduction into Vector)
(Primer 23: SEQ ID NO: 42)
5'-CTCGAGAACAATGGCGGATCTGAAGTCAAC-3'
(Primer 24: SEQ ID NO: 43)
5'-GGTACCTGTTTCTGTCTCTTGTAAATTTTGGC-3'
(Primer 25: SEQ ID NO: 44)
5'-CTCGAGAACAATGAGTTCAGTGAATTTGGG-3'
(Primer 26: SEQ ID NO: 45)
5'-GGATCCTTGTTTTGCCTGTGAGCGATGTAATTAGC-3'
(Primer 27: SEQ ID NO: 46)
5'-CTCGAGACAAATGGACACCACCGGCCGGCTCC-3'
(Primer 28: SEQ ID NO: 47)
5'-GGTACCACAGATGCAGCTTTAGACATATCTTTGC-3'
(Primer 29: SEQ ID NO: 48)
5'-CTCGAGACAAATGGACGAGGTTCGCCGGCGACC-3'
(Primer 30: SEQ ID NO: 49)
5'-GGTACCACGAAAGTTATTTTGGATACATCTTTTGC-3'
(Primer 31: SEQ ID NO: 50)
5'-AAGCTTACAAATGGATGTGCGCCGGCGACC-3'
(Primer 32: SEQ ID NO: 51)
5'-GGTACCACGGAAGAAGGCTTGGAAACAGC-3'
(Primer 33: SEQ ID NO: 52)
5'-AAGCTTACAAATGGATGCCCGCCGGCGACC-3'
(Primer 34: SEQ ID NO: 53)
5'-GGTACCACATTTACCTCAATAGAAGGCATTGTC-3'
(Primer 35: SEQ ID NO: 54)
5'-CTCGAGAACAATGGGTGAGGCTCCAGATGTCG-3'
(Primer 36: SEQ ID NO: 55)
5'-GGTACCTGACTCAACTTGTGAATCGTTTTCATGTC-3'
(Primer 37: SEQ ID NO: 56)
5'-CTCGAGCCAACAATGGAATTATACAACG-3'
(Primer 38: SEQ ID NO: 57)
5'-GGATCCTCTTTTAAGTATTCCTTATGTTTCTCC-3'
(Primer 47: SEQ ID NO: 70)
5'-CTCGAGCCAACAATGGAATTATACAACG-3'
(Primer 48: SEQ ID NO: 71)
5'-GGATCCTCTTTTAAGTATTCCTTATGTTTCTCC-3'
(Primer 39: SEQ ID NO: 58)
5'-CTCGAGAACAATGGCTGAAGAGGTGGAGG-3'
(Primer 40: SEQ ID NO: 59)
5'-GGATCCTGTGATGCCTCATCTCCAAACACC-3'
(Primer 43: SEQ ID NO: 64)
5'-CTCGAGAACAATGGCTGAAGACGAAGACAACC-3'
(Primer 44: SEQ ID NO: 65)
5'-GGATCCAAATTCTCTCCATAAAACACCTTAGC-3'
(Construction of Expression Vectors)
(Preparation of Expression Vector-Introduced Agrobacterium)
[Examples 1 to 11]
NAA: naphthaleneacetic acid;
2,4-D: 2,4-dichlorophenoxyacetic acid;
IBA: indolebutyric acid;
BA: benzyladenine;
KI: kinetin; and
para rubber tree: obtained from the Arboricultural Research Institute of the University of Tokyo Forests, Graduate School of Agricultural and Life Sciences, the University of Tokyo.
(Callus Induction (Induction Step))
(Infection Step and Coculture Step)
(Selective Culture Step)
(Formation of Adventitious Embryo and Shoot (Regeneration-inducing Step))
(Shoot Elongation (Elongation Step))
(Rooting (Rooting Step))
(Calculation of Polyisoprenoid Production and Measurement of Weight Average Molecular Weight of Polyisoprenoid)
[Comparative Examples 1 to 7]
Control | Example 1 | Comparative Example 1 | Example 2 | Comparative Example 2 | Example 3 | Comparative Example 3 | Example 4 | Comparative Example 4 | Example 5 | Comparative Example 5 | Example 6 | Comparative Example 6 | Example 7 | Comparative Example 7 | |
Promoter | - | HEV2.1 pro | 35S | HEV2.1 pro | 35S | HEV2.1 pro | 35S | HEV2.1 pro | 35S | HEV2.1 pro | 35S | HEV2.1 pro | 35S | HEV2.1 pro | 35S |
Protein encoded by gene introduced into transgenic plant | - | FPS | FPS | GGPS | GGPS | HMGR1 | HMGR1 | IPI | IPI | CPT1 | CPT1 | SRPP | SRPP | REF | REF |
Polyisoprenoid accumulation (% by mass) | 100 | 105 | 101 | 102 | 100 | 114 | 102 | 106 | 102 | 115 | 101 | 105 | 102 | 109 | 102 |
Weight average molecular weight (Mw) of polyisoprenoid | 8.8 × 105 | 9.0 × 105 | 8.8 × 105 | 9.1 × 105 | 8.8 × 105 | 1.2 ×106 | 8.9 × 105 | 8.8 × 105 | 8.8 × 105 | 1.4 × 106 | 9.0 × 105 | 9.1 × 105 | 8.7 × 105 | 9.1 × 105 | 8.8 × 105 |
HEV2.1 pro: the promoter of the gene encoding HEV2.1 (Hevein 2.1)
35S: cauliflower mosaic virus 35S promoter
FPS: farnesyl diphosphate synthase
GGPS: geranylgeranyl diphosphate synthase
HMGR1: 3-hydroxy-3-methylglutaryl CoA reductase 1
IPI: isopentenyl diphosphate isomerase
CPT1: cis-prenyltransferase 1
SRPP: Small Rubber Particle Protein
REF: Rubber Elongation Factor
(SEQUENCE LISTING FREE TEXT)
SEQ ID NO: 1: base sequence of promoter of gene encoding Hevein 2.1 derived from para rubber tree
SEQ ID NO: 2: base sequence of gene encoding farnesyl diphosphate synthase derived from para rubber tree
SEQ ID NO: 3: amino acid sequence of farnesyl diphosphate synthase derived from para rubber tree
SEQ ID NO: 4: base sequence of gene encoding geranylgeranyl diphosphate synthase derived from para rubber tree
SEQ ID NO: 5: amino acid sequence of geranylgeranyl diphosphate synthase derived from para rubber tree
SEQ ID NO: 6: base sequence of gene encoding 3-hydroxy-3-methylglutaryl CoA reductase 1 derived from para rubber tree
SEQ ID NO: 7: base sequence of gene encoding 3-hydroxy-3-methylglutaryl CoA reductase 3 derived from para rubber tree
SEQ ID NO: 8: base sequence of gene encoding 3-hydroxy-3-methylglutaryl CoA reductase 4 derived from para rubber tree
SEQ ID NO: 9: base sequence of gene encoding 3-hydroxy-3-methylglutaryl CoA reductase 5 derived from para rubber tree
SEQ ID NO: 10: amino acid sequence of 3-hydroxy-3-methylglutaryl CoA reductase 1 derived from para rubber tree
SEQ ID NO: 11: amino acid sequence of 3-hydroxy-3-methylglutaryl CoA reductase 3 derived from para rubber tree
SEQ ID NO: 12: amino acid sequence of 3-hydroxy-3-methylglutaryl CoA reductase 4 derived from para rubber tree
SEQ ID NO: 13: amino acid sequence of 3-hydroxy-3-methylglutaryl CoA reductase 5 derived from para rubber tree
SEQ ID NO: 14: base sequence of gene encoding isopentenyl diphosphate isomerase derived from para rubber tree
SEQ ID NO: 15: amino acid sequence of isopentenyl diphosphate isomerase derived from para rubber tree
SEQ ID NO: 16: base sequence of gene encoding cis-prenyltransferase 1 derived from para rubber tree
SEQ ID NO: 17: amino acid sequence of cis-prenyltransferase 1 derived from para rubber tree
SEQ ID NO: 18: base sequence of gene encoding Small Rubber Particle Protein derived from para rubber tree
SEQ ID NO: 19: amino acid sequence of Small Rubber Particle Protein derived from para rubber tree
SEQ ID NO: 20: Primer 1
SEQ ID NO: 21: Primer 2
SEQ ID NO: 22: Primer 3
SEQ ID NO: 23: Primer 4
SEQ ID NO: 24: Primer 5
SEQ ID NO: 25: Primer 6
SEQ ID NO: 26: Primer 7
SEQ ID NO: 27: Primer 8
SEQ ID NO: 28: Primer 9
SEQ ID NO: 29: Primer 10
SEQ ID NO: 30: Primer 11
SEQ ID NO: 31: Primer 12
SEQ ID NO: 32: Primer 13
SEQ ID NO: 33: Primer 14
SEQ ID NO: 34: Primer 15
SEQ ID NO: 35: Primer 16
SEQ ID NO: 36: Primer 17
SEQ ID NO: 37: Primer 18
SEQ ID NO: 38: Primer 19
SEQ ID NO: 39: Primer 20
SEQ ID NO: 40: Primer 21
SEQ ID NO: 41: Primer 22
SEQ ID NO: 42: Primer 23
SEQ ID NO: 43: Primer 24
SEQ ID NO: 44: Primer 25
SEQ ID NO: 45: Primer 26
SEQ ID NO: 46: Primer 27
SEQ ID NO: 47: Primer 28
SEQ ID NO: 48: Primer 29
SEQ ID NO: 49: Primer 30
SEQ ID NO: 50: Primer 31
SEQ ID NO: 51: Primer 32
SEQ ID NO: 52: Primer 33
SEQ ID NO: 53: Primer 34
SEQ ID NO: 54: Primer 35
SEQ ID NO: 55: Primer 36
SEQ ID NO: 56: Primer 37
SEQ ID NO: 57: Primer 38
SEQ ID NO: 58: Primer 39
SEQ ID NO: 59: Primer 40
SEQ ID NO: 60: base sequence of gene encoding Rubber Elongation Factor derived from para rubber tree
SEQ ID NO: 61: amino acid sequence of Rubber Elongation Factor derived from para rubber tree
SEQ ID NO: 62: Primer 41
SEQ ID NO: 63: Primer 42
SEQ ID NO: 64: Primer 43
SEQ ID NO: 65: Primer 44
SEQ ID NO: 66: base sequence of gene encoding cis-prenyltransferase 2 derived from para rubber tree
SEQ ID NO: 67: amino acid sequence of cis-prenyltransferase 2 derived from para rubber tree
SEQ ID NO: 68: Primer 45
SEQ ID NO: 69: Primer 46
SEQ ID NO: 70: Primer 47
SEQ ID NO: 71: Primer 48
SEQUENCE LISTING
<110> SUMITOMO RUBBER INDUSTRIES, LTD.
<120> VECTOR COMPRISING SPECIFIC PROMOTER AND GENE ENCODING SPECIFIC PROTEIN, TRANSGENIC PLANT INTO WHICH THE VECTOR HAS BEEN INTRODUCED, AND METHOD FOR IMPROVING POLYISOPRENOID PRODUCTION BY INTRODUCING THE VECTOR INTO PLANT
<130> SR1878
<160> 71
<170> PatentIn version 3.5
<210> 1
<211> 1680
<212> DNA
<213> Hevea brasiliensis
<400> 1
<210> 2
<211> 1029
<212> DNA
<213> Hevea brasiliensis
<400> 2
<210> 3
<211> 342
<212> PRT
<213> Hevea brasiliensis
<400> 3
<210> 4
<211> 1113
<212> DNA
<213> Hevea brasiliensis
<400> 4
<210> 5
<211> 370
<212> PRT
<213> Hevea brasiliensis
<400> 5
<210> 6
<211> 1728
<212> DNA
<213> Hevea brasiliensis
<400> 6
<210> 7
<211> 1761
<212> DNA
<213> Hevea brasiliensis
<400> 7
<210> 8
<211> 1821
<212> DNA
<213> Hevea brasiliensis
<400> 8
<210> 9
<211> 1581
<212> DNA
<213> Hevea brasiliensis
<400> 9
<210> 10
<211> 575
<212> PRT
<213> Hevea brasiliensis
<400> 10
<210> 11
<211> 586
<212> PRT
<213> Hevea brasiliensis
<400> 11
<210> 12
<211> 606
<212> PRT
<213> Hevea brasiliensis
<400> 12
<210> 13
<211> 526
<212> PRT
<213> Hevea brasiliensis
<400> 13
<210> 14
<211> 705
<212> DNA
<213> Hevea brasiliensis
<400> 14
<210> 15
<211> 234
<212> PRT
<213> Hevea brasiliensis
<400> 15
<210> 16
<211> 873
<212> DNA
<213> Hevea brasiliensis
<400> 16
<210> 17
<211> 290
<212> PRT
<213> Hevea brasiliensis
<400> 17
<210> 18
<211> 615
<212> DNA
<213> Hevea brasiliensis
<400> 18
<210> 19
<211> 204
<212> PRT
<213> Hevea brasiliensis
<400> 19
<210> 20
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 1
<400> 20
ctcaaggcta ccttattggg 20
<210> 21
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 2
<400> 21
ctcagcaatt gcaacacctg 20
<210> 22
<211> 27
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 3
<400> 22
ggtaccgcta ccttattggg aactacc 27
<210> 23
<211> 26
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 4
<400> 23
agatctaact cttcccattt cttccc 26
<210> 24
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 5
<400> 24
gaatccatgg cggatctgaa g 21
<210> 25
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 6
<400> 25
gtccatgtat ctggataccc 20
<210> 26
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 7
<400> 26
caagatgagt tcagtgaatt tggg 24
<210> 27
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 8
<400> 27
tgcattagtt ttgcctgtga gc 22
<210> 28
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 9
<400> 28
atttttacat ggacaccacc g 21
<210> 29
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 10
<400> 29
accagattcc cactaagatg c 21
<210> 30
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 11
<400> 30
tccatatatg gacgaggttc g 21
<210> 31
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 12
<400> 31
gcagctgtgt tacccttcag 20
<210> 32
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 13
<400> 32
cagtcgctcc aaaatggatg tgc 23
<210> 33
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 14
<400> 33
gattttctta ggaagaaggc ttgg 24
<210> 34
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 15
<400> 34
ctagctggtc tataatggat gcc 23
<210> 35
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 16
<400> 35
gaatcaattt acctcaatag aaggc 25
<210> 36
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 17
<400> 36
ttccaccatg ggtgaggctc c 21
<210> 37
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 18
<400> 37
tctcaactca acttgtgaat cg 22
<210> 38
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 19
<400> 38
atggaattat acaacggtga gagg 24
<210> 39
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 20
<400> 39
ttttaagtat tccttatgtt tctcc 25
<210> 40
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 21
<400> 40
tatggctgaa gaggtggagg 20
<210> 41
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 22
<400> 41
tgatgcctca tctccaaaca cc 22
<210> 42
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 23
<400> 42
ctcgagaaca atggcggatc tgaagtcaac 30
<210> 43
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 24
<400> 43
ggtacctgtt tctgtctctt gtaaattttg gc 32
<210> 44
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 25
<400> 44
ctcgagaaca atgagttcag tgaatttggg 30
<210> 45
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 26
<400> 45
ggatccttgt tttgcctgtg agcgatgtaa ttagc 35
<210> 46
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 27
<400> 46
ctcgagacaa atggacacca ccggccggct cc 32
<210> 47
<211> 34
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 28
<400> 47
ggtaccacag atgcagcttt agacatatct ttgc 34
<210> 48
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 29
<400> 48
ctcgagacaa atggacgagg ttcgccggcg acc 33
<210> 49
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 30
<400> 49
ggtaccacga aagttatttt ggatacatct tttgc 35
<210> 50
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 31
<400> 50
aagcttacaa atggatgtgc gccggcgacc 30
<210> 51
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 32
<400> 51
ggtaccacgg aagaaggctt ggaaacagc 29
<210> 52
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 33
<400> 52
aagcttacaa atggatgccc gccggcgacc 30
<210> 53
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 34
<400> 53
ggtaccacat ttacctcaat agaaggcatt gtc 33
<210> 54
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 35
<400> 54
ctcgagaaca atgggtgagg ctccagatgt cg 32
<210> 55
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 36
<400> 55
ggtacctgac tcaacttgtg aatcgttttc atgtc 35
<210> 56
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 37
<400> 56
ctcgagccaa caatggaatt atacaacg 28
<210> 57
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 38
<400> 57
ggatcctctt ttaagtattc cttatgtttc tcc 33
<210> 58
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 39
<400> 58
ctcgagaaca atggctgaag aggtggagg 29
<210> 59
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 40
<400> 59
ggatcctgtg atgcctcatc tccaaacacc 30
<210> 60
<211> 417
<212> DNA
<213> Hevea brasiliensis
<400> 60
<210> 61
<211> 138
<212> PRT
<213> Hevea brasiliensis
<400> 61
<210> 62
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 41
<400> 62
atggctgaag acgaagacaa cc 22
<210> 63
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 42
<400> 63
attctctcca taaaacacct tagc 24
<210> 64
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 43
<400> 64
ctcgagaaca atggctgaag acgaagacaa cc 32
<210> 65
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 44
<400> 65
ggatccaaat tctctccata aaacacctta gc 32
<210> 66
<211> 855
<212> DNA
<213> Hevea brasiliensis
<400> 66
<210> 67
<211> 284
<212> PRT
<213> Hevea brasiliensis
<400> 67
<210> 68
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 45
<400> 68
atggaattat acaacggtga gagg 24
<210> 69
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 46
<400> 69
ttttaagtat tccttatgtt tctcc 25
<210> 70
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 47
<400> 70
ctcgagccaa caatggaatt atacaacg 28
<210> 71
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> PRIMER 48
<400> 71
ggatcctctt ttaagtattc cttatgtttc tcc 33
a Hevein 2.1 encoding gene promoter; and
a gene, wherein the gene is functionally linked to the promoter, wherein the gene is at least one gene selected from the group consisting of a gene encoding farnesyl diphosphate synthase, a gene encoding geranylgeranyl diphosphate synthase, a gene encoding 3-hydroxy-3-methylglutaryl CoA reductase, a gene encoding isopentenyl diphosphate isomerase, a gene encoding cis-prenyltransferase, a gene encoding Small Rubber Particle Protein, and a gene encoding Rubber Elongation Factor,
wherein the Hevein 2.1 encoding gene promoter comprises any one of the following DNAs:
[A1] a DNA comprising the base sequence of base numbers 1 to 1680 represented by SEQ ID NO: 1;
[A2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1680 represented by SEQ ID NO: 1 under stringent conditions, and having a promoter activity for laticifer-specific gene expression; and
[A3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1680 represented by SEQ ID NO: 1, and having a promoter activity for laticifer-specific gene expression.
[B1] a DNA comprising the base sequence of base numbers 1 to 1029 represented by SEQ ID NO: 2;
[B2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1029 represented by SEQ ID NO: 2 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates or a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates; and
[B3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1029 represented by SEQ ID NO: 2, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates or a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates.
[C1] a DNA comprising the base sequence of base numbers 1 to 1113 represented by SEQ ID NO: 4;
[C2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1113 represented by SEQ ID NO: 4 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates, a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates, or a reaction using isopentenyl diphosphate and farnesyl diphosphate as substrates; and
[C3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1113 represented by SEQ ID NO: 4, and encoding a protein having an enzyme activity that catalyzes a reaction using isopentenyl diphosphate and dimethylallyl diphosphate as substrates, a reaction using isopentenyl diphosphate and geranyl diphosphate as substrates, or a reaction using isopentenyl diphosphate and farnesyl diphosphate as substrates.
[D1] a DNA comprising the base sequence of base numbers 1 to 1728 represented by SEQ ID NO: 6;
[D2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1728 represented by SEQ ID NO: 6 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1728 represented by SEQ ID NO: 6, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D4] a DNA comprising the base sequence of base numbers 1 to 1761 represented by SEQ ID NO: 7;
[D5] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1761 represented by SEQ ID NO: 7 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D6] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1761 represented by SEQ ID NO: 7, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D7] a DNA comprising the base sequence of base numbers 1 to 1821 represented by SEQ ID NO: 8;
[D8] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1821 represented by SEQ ID NO: 8 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D9] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1821 represented by SEQ ID NO: 8, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA;
[D10] a DNA comprising the base sequence of base numbers 1 to 1581 represented by SEQ ID NO: 9;
[D11] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 1581 represented by SEQ ID NO: 9 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA; and
[D12] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 1581 represented by SEQ ID NO: 9, and encoding a protein having an enzyme activity that catalyzes a reaction of reduction of 3-hydroxy-3-methylglutaryl CoA.
[E1] a DNA comprising the base sequence of base numbers 1 to 705 represented by SEQ ID NO: 14;
[E2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 705 represented by SEQ ID NO: 14 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of isomerization of isopentenyl diphosphate or dimethylallyl diphosphate; and
[E3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 705 represented by SEQ ID NO: 14, and encoding a protein having an enzyme activity that catalyzes a reaction of isomerization of isopentenyl diphosphate or dimethylallyl diphosphate.
[F1] a DNA comprising the base sequence of base numbers 1 to 873 represented by SEQ ID NO: 16;
[F2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 873 represented by SEQ ID NO: 16 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[F3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 873 represented by SEQ ID NO: 16, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds;
[F4] a DNA comprising the base sequence of base numbers 1 to 855 represented by SEQ ID NO: 66;
[F5] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 855 represented by SEQ ID NO: 66 under stringent conditions, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds; and
[F6] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 855 represented by SEQ ID NO: 66, and encoding a protein having an enzyme activity that catalyzes a reaction of cis-chain elongation of isoprenoid compounds.
[G1] a DNA comprising the base sequence of base numbers 1 to 615 represented by SEQ ID NO: 18;
[G2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 615 represented by SEQ ID NO: 18 under stringent conditions, and encoding a rubber particle-associated protein which is associated with rubber particles in latex; and
[G3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 615 represented by SEQ ID NO: 18, and encoding a rubber particle-associated protein which is associated with rubber particles in latex.
[H1] a DNA comprising the base sequence of base numbers 1 to 417 represented by SEQ ID NO: 60;
[H2] a DNA hybridizing to a DNA comprising a base sequence complementary to the base sequence of base numbers 1 to 417 represented by SEQ ID NO: 60 under stringent conditions, and encoding a rubber particle-associated protein which is associated with rubber particles in latex; and
[H3] a DNA comprising a base sequence having 80% or more sequence identity with the base sequence of base numbers 1 to 417 represented by SEQ ID NO: 60, and encoding a rubber particle-associated protein which is associated with rubber particles in latex.
einen Hevein 2.1 codierenden Genpromotor und
ein Gen, wobei das Gen funktionell mit dem Promotor verknüpft ist, wobei das Gen wenigstens ein Gen ist, welches aus der Gruppe ausgewählt ist, welche besteht aus einem Gen codierend Farnesyldiphosphatsynthase, ein Gen codierend Geranylgeranyldiphosphatsynthase, ein Gen codierend 3-Hydroxy-3-methylglutaryl-CoA-reduktase, ein Gen codierend Isopentenyldiphosphatisomerase, ein Gen codierend cis-Prenyltransferase, ein Gen codierend Kleines-Kautschuk-Partikel-Protein und ein Gen codierend Kautschukverlängerungsfaktor,
wobei der Hevein 2.1 codierende Genpromotor eine der nachfolgenden DNS'en enthält:
[A1] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 1680 wiedergegeben durch SEQ ID NO: 1,
[A2] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 1680 wiedergegeben durch SEQ ID NO:1 unter stringenten Bedingungen und mit einer Promotor-Aktivität für eine laticiferspezifische Genexpression und
[A3] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 1680 wiedergegeben durch SEQ ID NO: 1 und mit einer Promotoraktivität für laticiferspezifische Genexpression.
[B1] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 1029 wiedergegeben durch SEQ ID NO: 2,
[B2] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 1029 wiedergegeben durch SEQ ID NO: 2 unter stringenten Bedingungen und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion unter Verwendung von Isopentenyldiphosphat und Dimethylallyldiphosphat als Substrate oder eine Reaktion unter Verwendung von Isopentenyldiphosphat und Geranyldiphosphat als Substrate katalysiert, und
[B3] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 1029 wiedergegeben durch SEQ ID NO: 2 und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion unter Verwendung von Isopentenyldiphosphat und Dimethylallyldiphosphat als Substrate oder eine Reaktion unter Verwendung von Isopentenyldiphosphat und Geranyldiphosphat als Substrate katalysiert.
[C1] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 1113 wiedergegeben durch SEQ ID NO: 4,
[C2] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 1113 wiedergegeben durch SEQ ID NO: 4 unter stringenten Bedingungen und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion unter Verwendung von Isopentenyldiphosphat und Dimethylallyldiphosphat als Substrate, eine Reaktion unter Verwendung von Isopentenyldiphosphat und Geranyldiphosphat als Substrate oder eine Reaktion unter Verwendung von Isopentenyldiphosphat und Farnesyldiphosphat als Substrate katalysiert, sowie
[C3] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 1113 wiedergegeben durch SEQ ID NO: 4 und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion unter Verwendung von Isopentenyldiphosphat und Dimethylallyldiphosphat als Substrate, eine Reaktion unter Verwendung von Isopentenyldiphosphat und Geranyldiphosphat als Substrate oder eine Reaktion unter Verwendung von Isopentenyldiphosphat und Farnesyldiphosphat als Substrate katalysiert.
[D1] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 1728 wiedergegeben durch SEQ ID NO: 6,
[D2] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 1728 wiedergegeben durch SEQ ID NO: 6 unter stringenten Bedingungen und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion der Reduktion von 3-Hydroxy-3-methylglutaryl-CoA katalysiert,
[D3] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz der Basennummern 1 bis 1728 wiedergegeben durch SEQ ID NO: 6 und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion der Reduktion von 3-Hydroxy-3-methylglutaryl-CoA katalysiert,
[D4] eine DNS enthaltend die Basensequenz mit den Basennummern 1 bis 1761 wiedergegeben durch SEQ ID NO: 7,
[D5] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 1761 wiedergegeben durch SEQ ID NO: 7 unter stringenten Bedingungen und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion der Reduktion von 3-Hydroxy-3-methylglutaryl-CoA katalysiert,
[D6] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 1761 wiedergegeben durch SEQ ID NO: 7 und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion der Reduktion von 3-Hydroxy-3-methylglutaryl-CoA katalysiert,
[D7] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 1821 wiedergegeben durch SEQ ID NO: 8,
[D8] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 1821 wiedergegeben durch SEQ ID NO: 8 unter stringenten Bedingungen und codierend ein Protein mit einer Enzymaktivität, weiche eine Reaktion der Reduktion der 3-Hydroxy-3-methylglutaryl-CoA katalysiert,
[D9] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 1821 wiedergegeben durch SEQ ID NO. 8 und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion der Reduktion der 3-Hydroxy-3-methylglutaryl-CoA katalysiert,
[D10] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 1581 wiedergegeben durch SEQ ID NO: 9,
[D11] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 1581 wiedergegeben durch SEQ ID NO: 9 unter stringenten Bedingungen und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion der Reduktion von 3-Hydroxy-3-methylglutaryl-CoA katalysiert, sowie
[D12] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz der Basennummern 1 bis 1581 wiedergegeben durch SEQ ID NO. 9 und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion der Reduktion der 3-Hydroxy-3-methylglutaryl-CoA katalysiert.
[E1] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 705 wiedergegeben durch SEQ ID NO: 14,
[E2] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 705 wiedergegeben durch SEQ ID NO: 14 unter stringenten Bedingungen und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion der Isomerisation von Isopentenyldiphosphat oder Dimethylallyldiphosphat katalysiert, sowie
[E3] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 705 wiedergegeben durch SEQ ID NO: 14, und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion der Isomerisation von Isopentenyldiphosphat oder Dimethylallyldiphosphat katalysiert.
[F1] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 873 widergegeben durch SEQ ID NO: 16,
[F2] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 873 wiedergegeben durch SEQ ID NO. 16 unter stringenten Bedingungen und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion von cis-Kettenverlängerung von Isoprenoidverbindungen katalysiert,
[F3] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 873 wiedergebgeben durch SEQ ID NO. 16 und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion von cis-Kettenverlängerung von Isoprenoidverbindungen kataylsiert,
[F4] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 855 wiedergegeben durch SEQ ID NO. 66,
[F5] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 855 widergegeben durch SEQ ID NO. 66 unter stringenten Bedingungen und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion von cis-Kettenverlängerung von Isprenoidverbindungen katalysiert, sowie
[F6] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 855 wiedergegeben durch SEQ ID NO. 66 und codierend ein Protein mit einer Enzymaktivität, welche eine Reaktion von cis-Kettenverlängerung von Isprenoidverbindungen katalysiert.
[G1] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 615 wiedergegeben durch SEQ ID NO: 18,
[G2] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 615 wiedergegeben durch SEQ ID NO. 18 unter stringenten Bedingungen und codierend ein Kautschukpartikel-assoziiertes Protein, weiches mit Kautschukpartikeln in Latex assoziiert ist, sowie
[G3] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 615 wiedergegeben durch SEQ ID NO: 18 und codierend ein Kautschukpartikel-assoziiertes Protein, welches in Latex mit Kautschukpartikeln assoziiert ist.
[H1] eine DNS enthaltend eine Basensequenz mit den Basennummern 1 bis 417 wiedergegeben durch SEQ ID NO. 60,
[H2] eine DNS hybridisierend an eine DNS enthaltend eine Basensequenz komplementär zu der Basensequenz mit den Basennummern 1 bis 417 wiedergegeben durch SEQ ID NO. 60 unter stringenten Bedingungen und codierend ein Kautschukpartikel-assoziiertes Protein, welches in Latex mit Kautschukpartikeln assoziiert ist, sowie
[H3] eine DNS enthaltend eine Basensequenz mit 80 % oder mehr Sequenzidentität mit der Basensequenz mit den Basennummern 1 bis 417 wiedergegeben durch SEQ ID NO. 60 und codierend ein Kautschukpartikel-assoziiertes Protein, welches in Latex mit Kautschukpartikeln assoziiert ist.
un promoteur de gène codant pour l'Hévéine 2.1 ; et
un gène, le gène étant fonctionnellement lié au promoteur, le gène étant au moins un gène choisi dans le groupe consistant en un gène codant pour la farnésyl diphosphate synthase, un gène codant pour la géranylgéranyl diphosphate synthase, un gène codant pour la 3-hydroxy-3-méthylglutaryl CoA réductase, un gène codant pour l'isopentényl diphosphate isomérase, un gène codant pour la cis-prényltransférase, un gène codant pour la Protéine des Petites Particules de Caoutchouc, et un gène codant pour le Facteur d'Allongement du Caoutchouc,
dans lequel le promoteur de gène codant pour l'Hévéine 2.1 comprend l'un quelconque des ADN suivants :
[A1] un ADN comprenant une séquence de bases de numéros de bases 1 à 1680 représentée par SEQ ID NO: 1 ;
[A2] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 1680 représentée par SEQ ID NO: 1 dans des conditions stringentes, et ayant une activité de promoteur pour une expression de gènes spécifiques des laticifères ; et
[A3] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 1680 représentée par SEQ ID NO: 1, et ayant une activité de promoteur pour une expression de gènes spécifiques des laticifères.
[B1] un ADN comprenant la séquence de bases de numéros de bases 1 à 1029 représentée par SEQ ID NO: 2 ;
[B2] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 1029 représentée par SEQ ID NO: 2 dans des conditions stringentes, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction utilisant le diphosphate d'isopentényle et le diphosphate de diméthylallyle comme substrats ou une réaction utilisant le diphosphate d'isopentényle et le diphosphate de géranyle comme substrats ; et
[B3] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 1029 représentée par SEQ ID NO: 2, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction utilisant le diphosphate d'isopentényle et le diphosphate de diméthylallyle comme substrats ou une réaction utilisant le diphosphate d'isopentényle et le diphosphate de géranyle comme substrats.
[C1] un ADN comprenant la séquence de bases de numéros de bases 1 à 1113 représentée par SEQ ID NO: 4 ;
[C2] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 1113 représentée par SEQ ID NO: 4 dans des conditions stringentes, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction utilisant le diphosphate d'isopentényle et le diphosphate de diméthylallyle comme substrats, une réaction utilisant le diphosphate d'isopentényle et le diphosphate de géranyle comme substrats, ou une réaction utilisant le diphosphate d'isopentényle et le diphosphate de farnésyle comme substrats ; et
[C3] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 1113 représentée par SEQ ID NO: 4, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction utilisant le diphosphate d'isopentényle et le diphosphate de diméthylallyle comme substrats, une réaction utilisant le diphosphate d'isopentényle et le diphosphate de géranyle comme substrats, ou une réaction utilisant le diphosphate d'isopentényle et le diphosphate de farnésyle comme substrats.
[D1] un ADN comprenant la séquence de bases de numéros de bases 1 à 1728 représentée par SEQ ID NO: 6 ;
[D2] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 1728 représentée par SEQ ID NO: 6 dans des conditions stringentes, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction de réduction de 3-hydroxy-3-méthylglutaryl CoA ;
[D3] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 1728 représentée par SEQ ID NO: 6, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction de réduction de 3-hydroxy-3-méthylglutaryl CoA ;
[D4] un ADN comprenant la séquence de bases de numéros de bases 1 à 1761 représentée par SEQ ID NO: 7 ;
[D5] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 1761 représentée par SEQ ID NO: 7 dans des conditions stringentes, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction de réduction de 3-hydroxy-3-méthylglutaryl CoA ;
[D6] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 1761 représentée par SEQ ID NO: 7, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction de réduction de 3-hydroxy-3-méthylglutaryl CoA ;
[D7] un ADN comprenant la séquence de bases de numéros de bases 1 à 1821 représentée par SEQ ID NO: 8 ;
[D8] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 1821 représentée par SEQ ID NO: 8 dans des conditions stringentes, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction de réduction de 3-hydroxy-3-méthylglutaryl CoA ;
[D9] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 1821 représentée par SEQ ID NO: 8, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction de réduction de 3-hydroxy-3-méthylglutaryl CoA ;
[D10] un ADN comprenant la séquence de bases de numéros de bases 1 à 1581 représentée par SEQ ID NO: 9 ;
[D11] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 1581 représentée par SEQ ID NO: 9 dans des conditions stringentes, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction de réduction de 3-hydroxy-3-méthylglutaryl CoA ; et
[D12] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 1581 représentée par SEQ ID NO: 9, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction de réduction de 3-hydroxy-3-méthylglutaryl CoA.
[El] un ADN comprenant la séquence de bases de numéros de bases 1 à 705 représentée par SEQ ID NO: 14 ;
[E2] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 705 représentée par SEQ ID NO: 14 dans des conditions stringentes, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction d'isomérisation de diphosphate d'isopentényle ou de diphosphate de diméthylallyle ; et
[E3] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 705 représentée par SEQ ID NO: 14, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction d'isomérisation de diphosphate d'isopentényle ou de diphosphate de diméthylallyle.
[F1] un ADN comprenant la séquence de bases de numéros de bases 1 à 873 représentée par SEQ ID NO: 16 ;
[F2] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 873 représentée par SEQ ID NO: 16 dans des conditions stringentes, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction d'allongement de chaîne cis de composés isoprénoïdes ;
[F3] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 873 représentée par SEQ ID NO: 16, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction d'allongement de chaîne cis de composés isoprénoïdes ;
[F4] un ADN comprenant la séquence de bases de numéros de base 1 à 855 représentée par SEQ ID NO: 66 ;
[F5] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 855 représentée par SEQ ID NO: 66 dans des conditions stringentes, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction d'allongement de chaîne cis de composés isoprénoïdes ; et
[F6] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 855 représentée par SEQ ID NO: 66, et codant pour une protéine ayant une activité enzymatique qui catalyse une réaction d'allongement de chaîne cis de composés isoprénoïdes.
[G1] un ADN comprenant la séquence de bases de numéros de bases 1 à 615 représentée par SEQ ID NO: 18 ;
[G2] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 615 représentée par SEQ ID NO: 18 dans des conditions stringentes, et codant pour une protéine associée aux particules de caoutchouc qui est associée aux particules de caoutchouc dans un latex ; et
[G3] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 615 représentée par SEQ ID NO: 18, et codant pour une protéine associée aux particules de caoutchouc qui est associée aux particules de caoutchouc dans un latex.
[H1] un ADN comprenant la séquence de bases de numéros de bases 1 à 417 représentée par SEQ ID NO: 60 ;
[H2] un ADN s'hybridant à un ADN comprenant une séquence de bases complémentaire à la séquence de bases de numéros de bases 1 à 417 représentée par SEQ ID NO: 60 dans des conditions stringentes, et codant pour une protéine associée aux particules de caoutchouc qui est associée aux particules de caoutchouc dans un latex ; et
[H3] un ADN comprenant une séquence de bases ayant 80 % ou plus d'identité de séquence avec la séquence de bases de numéros de bases 1 à 417 représentée par SEQ ID NO: 60, et codant pour une protéine associée aux particules de caoutchouc qui est associée aux particules de caoutchouc dans un latex.
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