WO1998041643A1 - Expression control polynucleotides - Google Patents
Expression control polynucleotides Download PDFInfo
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- WO1998041643A1 WO1998041643A1 PCT/GB1998/000833 GB9800833W WO9841643A1 WO 1998041643 A1 WO1998041643 A1 WO 1998041643A1 GB 9800833 W GB9800833 W GB 9800833W WO 9841643 A1 WO9841643 A1 WO 9841643A1
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- Prior art keywords
- expression control
- polynucleotide
- control polynucleotide
- pollen
- expression
- Prior art date
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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)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
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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
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
- C12N15/8222—Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation
- C12N15/823—Reproductive tissue-specific promoters
- C12N15/8231—Male-specific, e.g. anther, tapetum, pollen
Definitions
- This invention relates to the fields of plant biotechnology and plant genetic engineering. In particular it relates to transgenic plant production and tissue-specific expression of introduced gene sequences in pollen cells.
- a promoter is a non-coding nucleotide sequence which controls the transcription of an adjacent nucleotide sequence.
- a number of promoters have been isolated from a wide variety of sources, including plants.
- the promoter is only active in particular tissue types expression of the heterologous sequence is restricted accordingly and this may be especially desirable in some circumstances.
- a number of plant-derived promoters have been isolated which activate expression of their companion nucleotide sequences only in pollen cells. Use of these pollen cell-specific promoters to activate genes encoding heterologous proteins has also been described [see CA 2021643] and may lead to the production of proteins not normally present in pollen cells. Such an approach may allow the expression of heterologous genes which encode for proteins able to render the plant male-sterile by ablation of pollen cells (for example if the proteins are toxic to the pollen cell) or to drive the production of antisense RNAs which interfere with the normal processes of pollen cell metabolism. Pollen cell-specific promoters can further be used to drive expression of proteins that are toxic to insects or other pests which consume pollen. These promoters can also be used to activate the expression of genes encoding proteins which will enhance the nutritional value of pollen.
- the number of pollen cell-specific promoters which have been well characterised is limited and different promoters exhibit a range of activities which cannot be predicted a priori and are difficult to quantify.
- the activity of a promoter isolated from one species of plant may also differ when the promoter is utilised in an heterologous species - such differences may be both in the tissue specificity and strength of the promoter and are more likely to occur with greater taxonomic distance between plant species.
- different promoters may be required to control expression of multiple genes since a gene silencing effect can occur if duplicate copies of the same promoter are used. The choice of promoter is therefore limited and has to be experimentally verified in the system under study.
- an invertase gene expression control polynucleotide a derivative, a functional equivalent, or a part thereof, which is pollen cell-specific.
- polynucleotide exhibits a distinct level of activity (or lack of activity) in pollen cells (ie in material ranging from developing pollen grain through to material derived from pollen) compared to the other tissue types of the transformed plant.
- control polynucleotide we mean any polynucleotide which is capable of affecting the expression of a gene.
- the term is intended to include promoters, enhancers and suppressors.
- derivative we mean a modified version of the expression control polynucleotide which exhibits substantial sequence homology to the original polynucleotide, for example which include nucleotide substitutions which have no effect on biological function.
- part we mean a deleted version of the expression control polynucleotide, which comprises at least a substantial portion of the original polynucleotide (for example at least 50% of said polynucleotide) .
- the preferred type of expression control polynucleotide is a promoter.
- the invertase gene promoter is preferably derived from a dicotyledon, such as potato.
- the expression control polynucleotide of the invention may comprise double- or single-stranded DNA or RNA.
- the invention also provides the use of the expression control polynucleotide described above to control expression of heterologous sequences.
- the expression control polynucleotide is used to drive pollen cell-specific expression of protein-encoding heterologous genes in plants eg monocotyledons or dicotyledons. Use of the expression control polynucleotide in this way in dicotyledons is preferred.
- the invention also provides a recombinant expression control polynucleotide comprising at least a part of a pollen cell-specific expression control polynucleotide as described above.
- the recombinant expression control polynucleotide of the invention is capable of specific expression of a heterologous sequence in pollen cells.
- the heterologous sequence expressed may encode a protein.
- RNA sequences which do not code for protein eg ribosomal RNA or anti-sense RNA may instead be transcribed from the heterologous sequence.
- the invention also provides a polynucleotide having the sequence set out in SEQ ID No 1, including derivatives, functional equivalents or parts thereof.
- the preferred polynucleotide is that shown in SEQ ID No 1 from nucleotides 3144-5396 and more preferably from nucleotides 3430-5349.
- the most preferred polynucleotide is the promoter in the 3430-5349 bp fragment.
- a deposit of genetic material containing the polynucleotide of SEQ ID No 1 was made at the National Collection of Type Cultures on 7 February 1997 under No NCTC 13013.
- the present invention also provides a recombinant nucleotide construct comprising an expression control polynucleotide according to the invention operably linked to a heterologous (preferably protein-encoding) polynucleotide.
- activation of the expression control polynucleotide may drive the expression of the heterologous polynucleotide, enabling production of the encoded protein. Since the expression control polynucleotide is tissue-specific, production of the protein will be limited to those tissues where the expression control polynucleotide is active.
- the present invention also provides a recombinant vector containing an expression control polynucleotide or a recombinant nucleotide construct as defined above.
- a method of producing a recombinant vector comprising ligating an expression control polynucleotide as described above into a suitable vector.
- a method of producing a transformed cell by transfecting a host cell using said recombinant vector forms another aspect of the invention.
- Suitable vectors and genetic modifications thereof are well- known in the art.
- the present invention also provides a transformed host cell containing a recombinant nucleotide construct or vector as defined above.
- the present invention also provides a transgenic organism (for example a transgenic plant) containing a recombinant nucleotide construct or a vector as defined above.
- the progeny (and seeds) of such transgenic organisms forms a further part of the invention.
- the present invention also provides a method for controlling the expression of a protein, said method comprising operably linking a polynucleotide sequence encoding said protein to an expression control polynucleotide of the invention.
- the method is especially useful for the expression of proteins in pollen.
- the protein expressed leads to sterility of the transformed plant.
- the invention also provides a method of controlling the expression of a heterologous polynucleotide in pollen, said method comprising operably linking said heterologous polynucleotide to an expression control polynucleotide of the invention.
- the promoter for the invertase gene of potato is expressed specifically in pollen to activate expression of any DNA sequences in the pollen of transgenic plants. Below we describe the isolation and characterisation of this promoter and how it has been used to express genes in pollen.
- Figure 1 Map of sequences detailed in text with restriction enzymes used in their cloning.
- Figure 2. whole anther from transgenic potato plant stained for GUS activity (GUS activity indicated by the dark areas ) .
- Figure 3 cross-section of anther as in Figure. 2 showing staining in individual pollen grains (pollen grains appear as dark spots).
- Figure 4 RT-PCR analysis showing a product of 374 bp indicating expression from the promoter in (a) floral and bud tissue, and (b) in excised anthers but not in the remainder of the floral tissue.
- Example A potato ( Solanum tuberos ⁇ m L. ) cv. Saturna genomic library consisting of a partial Sau3AI digest of genomic DNA cloned into AEMBL3 , was plated to yield 1 x 10 5 pfu which were screened with a radiolabelled carrot invertase cDNA fragment generated by reverse transcription-polymerase chain reaction (RT-PCR) using primers derived from a sequence of carrot cDNA (Sturm and Chrispeels, 1990).
- RT-PCR reverse transcription-polymerase chain reaction
- the primers were: Forward Primer: 5 '-AACGATCCAAATGGACCA-3 ' (SEQ ID No 2 ) Reverse Primer: 5 '-GAAAAAATCAGGACATTCCCA-3 ' (SEQ ID No 3) .
- Figure 1 It was determined, by homology to known invertase gene sequences, that the 5.4 kb of potato DNA (Figure 1) carried sequence of two invertase genes with the intergenic region constituting the promoter of the downstream gene.
- a 2.25 kb Hindlll-Xbal fragment (bp 3144-596; Figure 1) comprising the promoter, 3' end of the upstream gene and 5' end of the downstream gene was subcloned into pUC19 to yield plasmid pGF5211 (deposited as NCTC 13013). This fragment was also cloned into pBll01.3 to give plasmid pRM11.2 which was used as a vector for stable plant transformation.
- pRM11.2 the fragment is fused to the uidA gene from Escherichia coli and when the promoter is active in plants would drive the transcription of this gene to produce the bacteria enzyme -glucuronidase (GUS) .
- GUS bacteria enzyme -glucuronidase
- An internal AccI fragment of 1.9 kb (bp 3430-5349; Figure 1) derived from the 2.2 kb fragment was also cloned into pBI101.3 to generate plasmid pRM12.3 which was also used as a vector for stable plant transformation. This fragment was also fused to the uidA gene to drive 5-glucuronidase synthesis when active.
- a series of transgenic lines of potato (cv. Desiree) plants were generated by Agrobacterium tu efaciens- mediated transformation using pRM12.3 and pRM11.2 as a vector. Plants derived from the use of pRM12.3 as a vector were passed through one cycle of tuberisation then grown in a controlled environment until flowering occurred. The floral tissues including anthers, sepals , petals and ovules were separately analysed by a GUS histochemical assay performed at two pH values: pH 5 to assay for endogenous enzyme activity (the control) and pH 7 to detect the activity derived from the uidA ene activated by the invertase promoter.
- the invention describes a promoter sequence demonstrated to be active specifically in pollen of Solarium tuberosum.
- This promoter is likely to be active in pollen of other species of the Solanceae, and may be active in pollen of other plant species including those in which the production of male sterile plants for hybrid production is important eg Lycopersicon esculentum and the Brassicaceae. It is a unique sequence described with this activity in Solanum tuberosum, and for other plants it provides an alternative to the use of perviously isolated promoters . It has the advantage of its own characteristic activity profile and when used in heterologous species may escape problems such as gene silencing, which can compromise the use of homologous promoters . It has potential use in the genetic engineering of male sterile plants and lines for the restoration of fertility, for the production of proteins in pollen which are toxic to insect and other pests, or for the production of protein of enhanced nutritional value in pollen.
- TTCATCAATA TTTCAACCGA GGGGGTTGGG GGTTGGAGTG GGGTGGGATT TTTTTTAATG 1320
- AATAACTCAT AAAAAAAATA AATAAGAAGG ATTTATTTGA TATATATGTGTGTTGAACAG 2400
- TCCATCTAAA AGTCAGAGTT TTCTGAACTA TTTAACCTAG GTAAATCTTG GGCGTTAAAT 2760 AGGTAGATTA AAATTTGACA AATATGGTGC TTGGTCCGGG TCAGCAACTA TTCTACCAAA 2820
- AAATTCCGAC AAAAGAATTA TAAGGTTCTC ATGTGCTCAG ATGCTAGAAG GTTTGTTTCT 5580
- TCATCATTTC TAAATCATCT GTGCTTCTTC AATAGAGAAT CCTCAAACCA TTCAAACGAA 6240
- GAAGAGTAAA AAATAAAAAA ATATTAAATA AACTAGTTCG AAAATTCTAA CGTAATAAAA 6360
- CTCTATTGAA CATCTAGGCG GAGACGTTCA TTCGCCTTCC TGTTTAAAAA TGGAATGATT 8160
- TTCTCTACGA CAATATCGTT TCTTACTATT CTTTTAGGGC TTTTCGCCCT AGGGATATAG 8820 TATATATGAT GTTATAGCAA AGAATGATAA GAAAATCCCG AAAAGCGGGA TCCCTATATC 8880
- ATATATACTC CAAATTTCAC GCCATATATA AATAATGTGG AAAAAACTTA TAATTTTCAA 8940
Abstract
There is provided an expression control polynucleotide of an invertase gene, which may be operably limited to a heterologous polynucleotide. Optionally the expression control polynucleotide and heterologous polynucleotide construct is transfected into host cells or organisms. Preferably the construct is used to produce a transgenic plant and the expression control polynucleotide is pollen cell-specific. A suitable expression control polynucleotide is as set out in SEQ ID No 1, especially nucleotides 3430-5349 thereof. Desirably the protein expressed by the heterologous polynucleotide causes male sterility in the transgenic plants.
Description
"Expression Control Polynucleotides"
This invention relates to the fields of plant biotechnology and plant genetic engineering. In particular it relates to transgenic plant production and tissue-specific expression of introduced gene sequences in pollen cells.
A promoter is a non-coding nucleotide sequence which controls the transcription of an adjacent nucleotide sequence. A number of promoters have been isolated from a wide variety of sources, including plants. In certain applications it is desirable to genetically engineer a construct which comprises a promoter operatively linked to a heterologous nucleotide sequence such that the promoter controls expression of the heterologous sequence in the host cell transformed with that construct. Where the promoter is only active in particular tissue types expression of the heterologous sequence is restricted accordingly and this may be especially desirable in some circumstances.
A number of plant-derived promoters have been isolated which activate expression of their companion nucleotide sequences only in pollen cells. Use of these pollen
cell-specific promoters to activate genes encoding heterologous proteins has also been described [see CA 2021643] and may lead to the production of proteins not normally present in pollen cells. Such an approach may allow the expression of heterologous genes which encode for proteins able to render the plant male-sterile by ablation of pollen cells (for example if the proteins are toxic to the pollen cell) or to drive the production of antisense RNAs which interfere with the normal processes of pollen cell metabolism. Pollen cell-specific promoters can further be used to drive expression of proteins that are toxic to insects or other pests which consume pollen. These promoters can also be used to activate the expression of genes encoding proteins which will enhance the nutritional value of pollen.
However, the number of pollen cell-specific promoters which have been well characterised is limited and different promoters exhibit a range of activities which cannot be predicted a priori and are difficult to quantify. The activity of a promoter isolated from one species of plant may also differ when the promoter is utilised in an heterologous species - such differences may be both in the tissue specificity and strength of the promoter and are more likely to occur with greater taxonomic distance between plant species. In addition different promoters may be required to control expression of multiple genes since a gene silencing effect can occur if duplicate copies of the same promoter are used. The choice of promoter is therefore limited and has to be experimentally verified in the system under study.
According to the present invention there is provided an invertase gene expression control polynucleotide, a
derivative, a functional equivalent, or a part thereof, which is pollen cell-specific.
By "pollen cell-specific" we mean that the expression control polynucleotide exhibits a distinct level of activity (or lack of activity) in pollen cells (ie in material ranging from developing pollen grain through to material derived from pollen) compared to the other tissue types of the transformed plant.
By "expression control polynucleotide" we mean any polynucleotide which is capable of affecting the expression of a gene. The term is intended to include promoters, enhancers and suppressors.
By "functional equivalent" we mean any variation of the expression control polynucleotide which exhibits substantially the same functional properties of the original polynucleotide.
By "derivative" we mean a modified version of the expression control polynucleotide which exhibits substantial sequence homology to the original polynucleotide, for example which include nucleotide substitutions which have no effect on biological function.
By "part" we mean a deleted version of the expression control polynucleotide, which comprises at least a substantial portion of the original polynucleotide (for example at least 50% of said polynucleotide) .
The preferred type of expression control polynucleotide is a promoter.
The invertase gene promoter is preferably derived from
a dicotyledon, such as potato.
The expression control polynucleotide of the invention may comprise double- or single-stranded DNA or RNA.
The invention also provides the use of the expression control polynucleotide described above to control expression of heterologous sequences. Optionally the expression control polynucleotide is used to drive pollen cell-specific expression of protein-encoding heterologous genes in plants eg monocotyledons or dicotyledons. Use of the expression control polynucleotide in this way in dicotyledons is preferred.
The invention also provides a recombinant expression control polynucleotide comprising at least a part of a pollen cell-specific expression control polynucleotide as described above. The recombinant expression control polynucleotide of the invention is capable of specific expression of a heterologous sequence in pollen cells. The heterologous sequence expressed may encode a protein. Alternatively RNA sequences which do not code for protein (eg ribosomal RNA or anti-sense RNA) may instead be transcribed from the heterologous sequence.
The invention also provides a polynucleotide having the sequence set out in SEQ ID No 1, including derivatives, functional equivalents or parts thereof. The preferred polynucleotide is that shown in SEQ ID No 1 from nucleotides 3144-5396 and more preferably from nucleotides 3430-5349. The most preferred polynucleotide is the promoter in the 3430-5349 bp fragment.
A deposit of genetic material containing the
polynucleotide of SEQ ID No 1 was made at the National Collection of Type Cultures on 7 February 1997 under No NCTC 13013.
The present invention also provides a recombinant nucleotide construct comprising an expression control polynucleotide according to the invention operably linked to a heterologous (preferably protein-encoding) polynucleotide.
Thus, activation of the expression control polynucleotide may drive the expression of the heterologous polynucleotide, enabling production of the encoded protein. Since the expression control polynucleotide is tissue-specific, production of the protein will be limited to those tissues where the expression control polynucleotide is active.
The present invention also provides a recombinant vector containing an expression control polynucleotide or a recombinant nucleotide construct as defined above.
According to the present invention there is also provided a method of producing a recombinant vector, said method comprising ligating an expression control polynucleotide as described above into a suitable vector. A method of producing a transformed cell by transfecting a host cell using said recombinant vector forms another aspect of the invention. Suitable vectors and genetic modifications thereof are well- known in the art.
The present invention also provides a transformed host cell containing a recombinant nucleotide construct or vector as defined above.
The present invention also provides a transgenic organism (for example a transgenic plant) containing a recombinant nucleotide construct or a vector as defined above. The progeny (and seeds) of such transgenic organisms forms a further part of the invention.
The present invention also provides a method for controlling the expression of a protein, said method comprising operably linking a polynucleotide sequence encoding said protein to an expression control polynucleotide of the invention. The method is especially useful for the expression of proteins in pollen. Preferably the protein expressed leads to sterility of the transformed plant.
Thus the invention also provides a method of controlling the expression of a heterologous polynucleotide in pollen, said method comprising operably linking said heterologous polynucleotide to an expression control polynucleotide of the invention.
In one embodiment the promoter for the invertase gene of potato is expressed specifically in pollen to activate expression of any DNA sequences in the pollen of transgenic plants. Below we describe the isolation and characterisation of this promoter and how it has been used to express genes in pollen.
The present invention will now be further described with reference to the Example and accompanying Figures in which:
Figure Legends
Figure 1. Map of sequences detailed in text with restriction enzymes used in their cloning.
Figure 2. whole anther from transgenic potato plant stained for GUS activity (GUS activity indicated by the dark areas ) .
Figure 3. cross-section of anther as in Figure. 2 showing staining in individual pollen grains (pollen grains appear as dark spots).
Figure 4. RT-PCR analysis showing a product of 374 bp indicating expression from the promoter in (a) floral and bud tissue, and (b) in excised anthers but not in the remainder of the floral tissue.
Example A potato ( Solanum tuberosυm L. ) cv. Saturna genomic library, consisting of a partial Sau3AI digest of genomic DNA cloned into AEMBL3 , was plated to yield 1 x 105 pfu which were screened with a radiolabelled carrot invertase cDNA fragment generated by reverse transcription-polymerase chain reaction (RT-PCR) using primers derived from a sequence of carrot cDNA (Sturm and Chrispeels, 1990).
The primers were: Forward Primer: 5 '-AACGATCCAAATGGACCA-3 ' (SEQ ID No 2 ) Reverse Primer: 5 '-GAAAAAATCAGGACATTCCCA-3 ' (SEQ ID No 3) .
Hybridisation conditions of 5 x SSC at 65°C were utilised with subsequent low stringency washing of filters in 2 x SSC at 65°C. After three rounds of screening two positive clones were obtained plaque pure. DNA was purified from one positive clone, AGF5, which was shown to contain an insert of approximately 23 kb of potato DNA. This cloned potato DNA was
digested with Xbal and Sail, and fragments cloned into pUC19. One subclone, named pGF521, contained 5.4 kb of the potato DNA. A complete DNA sequence of this fragment is presented (SEQ ID No 1). It was determined, by homology to known invertase gene sequences, that the 5.4 kb of potato DNA (Figure 1) carried sequence of two invertase genes with the intergenic region constituting the promoter of the downstream gene. A 2.25 kb Hindlll-Xbal fragment (bp 3144-596; Figure 1) comprising the promoter, 3' end of the upstream gene and 5' end of the downstream gene was subcloned into pUC19 to yield plasmid pGF5211 (deposited as NCTC 13013). This fragment was also cloned into pBll01.3 to give plasmid pRM11.2 which was used as a vector for stable plant transformation. In pRM11.2 the fragment is fused to the uidA gene from Escherichia coli and when the promoter is active in plants would drive the transcription of this gene to produce the bacteria enzyme -glucuronidase (GUS) . An internal AccI fragment of 1.9 kb (bp 3430-5349; Figure 1) derived from the 2.2 kb fragment was also cloned into pBI101.3 to generate plasmid pRM12.3 which was also used as a vector for stable plant transformation. This fragment was also fused to the uidA gene to drive 5-glucuronidase synthesis when active.
A series of transgenic lines of potato (cv. Desiree) plants were generated by Agrobacterium tu efaciens- mediated transformation using pRM12.3 and pRM11.2 as a vector. Plants derived from the use of pRM12.3 as a vector were passed through one cycle of tuberisation then grown in a controlled environment until flowering occurred. The floral tissues including anthers, sepals , petals and ovules were separately analysed by a GUS histochemical assay performed at two pH values: pH 5 to assay for endogenous enzyme activity (the control)
and pH 7 to detect the activity derived from the uidA ene activated by the invertase promoter. A strong blue staining, detected only at pH 7 and thus indicative of bacterial GUS derived from expression of uidA driven by the invertase promoter, was observed only in pollen cells (see dark areas of Figures 2 and 3) and in no other tissues of the flower or elsewhere throughout the plant, while in control untransformed plants only a light background of blue staining was observed in pollen cells. Prior to the GUS histochemical analysis an analysis using RT-PCR to detect expression from the native promoter driving its invertase gene had detected expression only in floral and bud tissue with no expression observed in source and sink leaf (Figure 4a), stem, root or tuber. A subsequent RT-PCR analysis detected expression only in the pollen-containing anthers and not elsewhere throughout the flower (Figure 4b) . We conclude that the activity of this promoter is restricted to pollen.
The recombinant DNA procedure utilised were as described by Sambrook et al (1989). Plant tissue culture and transformation protocols were as detailed by Hedley (1995). Histochemical assay of GUS was performed as indicated by Jefferson (1987).
The invention describes a promoter sequence demonstrated to be active specifically in pollen of Solarium tuberosum. This promoter is likely to be active in pollen of other species of the Solanceae, and may be active in pollen of other plant species including those in which the production of male sterile plants for hybrid production is important eg Lycopersicon esculentum and the Brassicaceae. It is a unique sequence described with this activity in Solanum tuberosum, and for other plants it provides an
alternative to the use of perviously isolated promoters . It has the advantage of its own characteristic activity profile and when used in heterologous species may escape problems such as gene silencing, which can compromise the use of homologous promoters . It has potential use in the genetic engineering of male sterile plants and lines for the restoration of fertility, for the production of proteins in pollen which are toxic to insect and other pests, or for the production of protein of enhanced nutritional value in pollen.
References Hedley (1995). PhD. Thesis, University of Dundee. Jefferson (1987). Plant Molecular Biology Reporter 5, 387-405. Sambrook et al. (1989). Molecular cloning A Laboratory Manual. Second edition. Cold spring Harbor Laboratory Press, Cold Spring Harbor, New York. Sturn & Chrispeels (1990). The Plant Cell 2, 1107- 1119.
SEQUENCE LISTING
(1) GENERAL INFORMATION:
(l) APPLICANT:
(A) NAME: SCOTTISH CROP RESEARCH INSTITUTE
(B) STREET: INVERGOWRIE
(C) CITY: DUNDEE
(E) COUNTRY: UK
(F) POSTAL CODE (ZIP) : DD2 5DA
(G) TELEPHONE: 01382 562731 (H) TELEFAX: 01382 562426
(u| TITLE OF INVENTION: EXPRESSION CONTROL POLYNUCLEOTIDES
(in) NUMBER OF SEQUENCES: 3
(IV) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk
(B) COMPUTER: IBM PC compatible
(C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: Patentln Release #1.0, Version #1.30 (EPO)
(2) INFORMATION FOR SEQ ID NO: 1:
(l) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 10811 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: double
(D) TOPOLOGY: linear
(n) MOLECULE TYPE: DNA (genomic)
(vi) ORIGINAL SOURCE:
(A) ORGANISM: Solanum tubersum
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:
AAGCCTAGTT CGACCTGCAG TCAACGGATC TTTATAGCTA CATATATATA AGATTGATCA 60
TTCTTGATAA GCTGGACGTC AGTTGCCTAG AAATATCGAT GTATATATAT TCTAACTAGT 120
AAGAACTATT TTCAAAATTA TGTATACATA CACACACATA CATAATTATG TGGTTCATTT 180
GTGTTAGTTA AAGTTTTAAT ACATATGTAT GTGTGTGTAT GTATTAATAC ACCAAGTAAA 240
CACAATCAAT TCTATTATTC AGTAGTCAGT ATTCATTTTT GAAATGTAAT TAATTTAAAT 300
TTGTGTCTAA AGATAATAAG TCATCAGTCA TAAGTAAAAA CTTTACATTA ATTAAATTTA 360
AACACAGATA ATATTCTATT TTGGAGAACA AAATCGCTCA TGATCAACAA TCGATGACTC 420
AATTTTTAAT TATAAGATAA AACCTCTTGT TTTAGCGAGT ACTAGTTGTT AGCTACTGAG 480
TTAAAAATTA ATTTAAATTC GAAATTAGAT TAATTATTAT GGCAAGACAA TTACAAGGCT 540
AAGGTTTTGT TAAATTTAAG CTTTAATCTA ATTAATAATA CCGTTCTGTT AATGTTCCGA 600
TTCCAAAACG ATAAGAATGT GCAAAAGAGA AAAAGAAACA TGAAATATAT GAAAAAGTTC 660
TTTTAACCTC TATTCTTACA CGTTTTCTCT TTTTCTTTGT ACTTTATATA CTTTTTCAAG 720
AAAATTGGAA AGATTTTGGC CATGGAATTA AGGTGAAAAT TAATTTGTTG GAGGCACCCT 780
TTATATTCCT TCTAAAACCG GTACCTTAAT TCCACTTTTA ATTAAACAAC CTCCGTGGGA 840
AATATAAGGC CTTGGCATTT TCTTCTCCCT TATATTTTTT CCTTCTAAAT TATTATTATT 900
ATTTTTATTG GAACCGTAAA AGAAGAGGGA ATATAAAAAA GGAAGATTTA ATAATAATAA 960
TAAAAATAAA TTATTATTAT TATTATTAAG TGTTGAAATA TAGTGACATT TCATACATAC 1020
TCACATATTT AATAATAATA ATAATAATTC ACAACTTTAT ATCACTGTAA AGTATGTATG 1080
AGTGTATAAG TGTACATTTA ATATGTAGGT CTTATATTAA TTAAAACTTG CCAAACATAT 1140
TGTCTCTTAC ACATGTAAAT TATACATCCA GAATATAATT AATTTTGAAC GGTTTGTATA 1200
ACAGAGAATT AAAGTTGGCT CCCCCAACCC CCAACCTCAC CCCACCCTAA AAAAAATTAC 1260
TTCATCAATA TTTCAACCGA GGGGGTTGGG GGTTGGAGTG GGGTGGGATT TTTTTTAATG 1320
AAGTAGTTAT TCGTTTTTTC ATGTGACACA ATTTTCTTAT TTAATTTGTT ATCTAAAGAA 1380
TGACTATTTA AGCAAAAAAG TACACTGTGT TAAAAGAATA AATTAAACAA TAGATTTCTT 1440
ACTGATAAAT ATATATTTAA ACCTGTACTT ATCATTCTAC ATGATATATA TATATATATA 1500
TATATATATA TATATAAATT TGGACATGAA TAGTAAGATG TACTATATAT ATATATATAT 1560
ATATATATAA TGTATATATA TGTGTGTGTG TGTTTTTATA GTCACAATAA ATCTCATGAT 1620
ATGTTTCAGT ACATATATAT ACACACACAC ACAAAAATAT CAGTGTTATT TAGAGTACTA 1680
TACAAAGTCA TCATAATTTT CAAATATTTT TTACTTTATT TTTTAAATTA CGTATTAAAT 1740
CAAATTACGT AGTATTAAAA GTTTATAAAA AATGAAATAA AAAATTTAAT GCATAATTTA 1800
GTTTAATGCC CATATAAATT AAAATGATTC AACTAATTAG TCATTTTTGT ATTTCCTACA 1860
TTTCTGTGTG GTATATTTAA TTTTACTAAG TTGATTAATC AGTAAAAACA TAAAGGATGT 1920
AAAGACACAT CACCTTTGAT TTGTTAATTA TTATAGTATT TGATTATTTC TTAATCATTG 1980
ATTAATTATA GTGGAAACTA AACAATTAAT AATATCATAA ACTAATAAAG AATTAGTAAC 2040
TAATTAATAG TAATGCTTTC TATTGTCTTC ACTAATCATT TCTTTTTTTA AAATTCTTTT 2100
GTTCTTATAC ATTACGAAAG ATAACAGAAG TGATTAGTAA AGAAAAAAAT TTTAAGAAAA 2160
CAAGAATATT GGTCTCTCTT CATTGGATAC CTTTCAAATA TACAAAGACC CTAATGGTGA 2220
GTTAGATTAA CCAGAGAGAA GTAACCTATG GAAAGTTTAT ATGTTTCTGG GATTACCACT 2280
CAATCTAATA TTTTTTTTAT TTATTCTTCC TAAATAAACT ATATATACAC ACAACTTGTC 2340
AATAACTCAT AAAAAAAATA AATAAGAAGG ATTTATTTGA TATATATGTG TGTTGAACAG 2400
TTATTGAGTA TTATATTATT GACTATCATT TTATTTTTCA ACAGCACCAA TGTATTATAA 2460
TGGAGTA AT AATATAATAA CTGATAGTAA AATAAAAAGT TGTCGTGGTT ACATAATATT 2520
ACCTCATATT CATTTATTCT ATCAATACAA TCCAAAAGGA TCAGTATGGG GCAATATTGT 2580
TTGGGCTCAA GTAAATAAGA TAGTTATGTT AGGTTTTCCT AGTCATACCC CGTTATAACA 2640
AACCCGAGTT TCAGTCTCAA AAGACTTGAT AAATTGGATC CATTTAGAAC CCGCAATTTA 2700
TCCATCTAAA AGTCAGAGTT TTCTGAACTA TTTAACCTAG GTAAATCTTG GGCGTTAAAT 2760
AGGTAGATTA AAATTTGACA AATATGGTGC TTGGTCCGGG TCAGCAACTA TTCTACCAAA 2820
TAACAAGCCT TTTAAACTGT TTATACCACG AACCAGGCCC AGTCGTTGAT AAGATGGTTT 2880
ATTGTTCGGC GTTATCTTAT ACACCGGAGT AGTAGATTCC CATGATTCTC AAGTTCAAAA 2940
TTATGCAATG CAATAGAATA TGTGGCCTCA TCATCTAAGG GTACTAAGAG TTCAAGTTTT 3000
AATACGTTAC CCGGCTAACT TGTCTGATCC ATTTCTTCGT AAATGGATCA AACCTAATAA 3060
CAACCCGTTG GGCCGATTGA ACAGACTAGG TAAAGAAGCA TTTACCTAGT TTGGATTATT 3120
GTTGGGCAAG ATTGTACCTG ACAATAGCAT CAACAAAACC AAATTTCGTG ATCCAACAAC 3180
CGCATGGATC TAACATGGAC TGTTATCGTA GTTGTTTTGG TTTAAAGCAC TAGGTTGTTG 3240
GCGTACCTAG GGCCAAGATG GGCTTTGGAG AATTGTAATA GGAAGTATGA GAAAACATAG 3300
AGGGATGGCC CCGGTTCTAC CCGAAACCTC TTAACATTAT CCTTCATACT CTTTTGTATC 3360
TCCCTACCGA TTATTGTATA GAAGTAGAGA CTTCATTAAA TGGGCCAAAG CCCAACATCC 3420
ACTTCATTCT AATAACATAT CTTCATCTCT GAAGTAATTT ACCCGGTTTC GGGTTGTAGG 3480
TGAAGTAAGA TCTCCTCATA CTGGAAATTG GGAATGTCCT GATTTTTTTC CTGTATCATT 3540
AAAAAATACT AGAGGAGTAT GACCTTTAAC CCTTACAGGA CTAAAAAAAG GACATAGTAA 3600
TTTTTTATGA AATGGCTTAG ATGCATCGTA TCGCGGAAAA AATGTCAAAT ATGTCCTTAA 3660
GAATAGCCTT TTACCGAATC TACGTAGCAT AGCGCCTTTT TTACAGTTTA TACAGGAATT 3720
CTTATCGGAT GATGTTAATA GGTTTGAGTA TTACACTATT GGTATGTATG ACACCAAAAA 3780
AGATAGGTAA CTACAATTAT CCAAACTCAT AATGTGATAA CCATACATAC TGTGGTTTTT 3840
TCTATCCATC ATTCCTGATA ACAATTCTAT CGATGGTTCG AAGGGATTGA GGCTTGACTA 3900
TGGGAATTTG TAAGGACTAT TGTTAAGATA GCTACCAAGC TTCCCTAACT CCGAACTGAT 3960
ACCCTTAAAC TATGCATCTA AATCATTCTA TGACCCTATG AAGAATCGAA GAATTGTATG 4020
GGGTTGGACG ATACGTAGAT TTAGTAAGAT ACTGGGATAC TTCTTAGCTT CTTAACATAC 4080
CCCAACCTGA AATGAATCAG ATGTTTTACC TGACGATGAA ATTAAGAAAG GATGGGCTGG 4140
AATTCAAGCT TTACTTAGTC TACAAAATGG ACTGCTACTT TAATTCTTTC CTACCCGACC 4200
TTAAGTTCGT ATTCCGCGTA AAGTATGGCT CGACCCTAGT GGTAAACAAC TGATTCAATG 4260
GCCTATTGAA TAAGGCGCAT TTCATACCGA GCTGGGATCA CCATTTGTTG ACTAAGTTAC 4320
CGGATAACTA GAATTAGAAA CATTAAGAAA GCAAAAGATT ATTCAATTGA ACAACAAGAA 4380
GTTGAGCAAT CTTAATCTTT GTAATTCTTT CGTTTTCTAA TAAGTTAACT TGTTGTTCTT 4440
CAACTCGTTG GGAGAAATGT TTGAAGTTAA AGGAATCTCA GCATCACAGG TTTTAACTTT 4500
TCCTTATTAC CCTCTTTACA AACTTCAATT TCCTTAGAGT CGTAGTGTCC AAAATTGAAA 4560
AGGAATAATA ACTATAGTCT TTTAAATATC ACAACAAAAA GTAATTTTAA TTAGTATACA 4620
ATGTATAATT TGATATCAGA AAATTTATAG TGTTGTTTTT CATTAAAATT AATCATATGT 4680
TACATATTAC AATGTATAAC TATTATATCA ATTGCATATG ATAGGTCTAT ATATAGTAGA 4740
TTAATTATTG TTACATATTG ATAATATAGT TAACGTATAC TATCCAGATA TATATCATCT 4800
AATTAATAAT ATTCACTTGT TATATGTATA TCGGTGATAA TTATAAATTG TTGCAATTGA 4860
ACGAGTGAAA TAAGTGAACA ATATACATAT AGCCACTATT AATATTTAAC AACGTTAACT 4920
TGCTCACTTA TATTTTTGGT TAGTCGGGAA AAAAATTTGA CTAGGGCTAG GGCTAAATAA 4980
ACTTGTGCGT ATAAAAACCA ATCAGCCCTT TTTTTAAACT GATCCCGATC CCGATTTATT 5040
TGAACACGCT TAGCTTCTTC TCATCTTACA TTTCTTTGGT TGCAGGCTGA TATTGAAGTG 5100
TCGTTCTCTA ATCGAAGAAG AGTAGAATGT AAAGAAACCA ACGTCCGACT ATAACTTCAC 5160
AGCAAGAGAT TTTCAAGTTT GAACAAGGCC GAACAATTTG ATCCTAAATG GGCCGACCTT 5220
TATGCCCAAA AAAGTTCAAA CTTGTTCCGG CTTGTTAAAC TAGGATTTAC CCGGCTGGAA 5280
ATACGGGTTG ATGTTTGTGC CATAAAGGGT TCGACTATCC AAGGTGGGCT TGGACCATTT 5340
GGGCTTGTGC TACAAACACG GTATTTCCCA AGCTGATAGG TTCCACCCGA ACCTGGTAAA 5400
CCCGAACACA CATTAGCTTC TAAAAACTTG GAAGAATACA CACCTGTTTT CTTTCGAGTG 5460
TTTAAGGCTT GTAATCGAAG ATTTTTGAAC CTTCTTATGT GTGGACAAAA GAAAGCTCAC 5520
AAATTCCGAC AAAAGAATTA TAAGGTTCTC ATGTGCTCAG ATGCTAGAAG GTTTGTTTCT 5580
TCAATTGAAG TTTTCTTAAT ATTCCAAGAG TACACGAGTC TACGATCTTC CAAACAAAGA 5640
AGTTAACTTT TAGTTGTAAT GATCATAGTT TACATAGTTA ACTCAATATA GGAGCACAAA 5700
ATTTGAGTAA ATCAACATTA CTAGTATCAA ATGTATCAAT TGAGTTATAT CCTCGTGTTT 5760
TAAACTCATA ATCAAGAATT ATAATGACCC GACTTTGATA TCATGATAAG AAATACATCT 5820
ACTTATCGAT TAGTTCTTAA TATTACTGGG CTGAAACTAT AGTACTATTC TTTATGTAGA 5880
TGAATAGCTT TGTTTATTAG TGTCATTAAA AAACTCTAAC CTTGTTTAGT TTCTTTATTA 5940
ATGAGCAGAA ACAAATAATC ACAGTAATTT TTTGAGATTG GAACAAATCA AAGAAATAAT 6000
TACTCGTCTT CTACCATGAG ACAAAATGAA GCAATGTACA AGCCCTCATT TGCTGGATAT 6060
GTAGATGTAA GATGGTACTC TGTTTTACTT CGTTACATGT TCGGGAGTAA ACGACCTATA 6120
CATCTACATG ATTTAGTAGA CACGAAGAAG TTATCTCTTA GGAGTTTGGT AAGTTTGCTT 6180
TCATCATTTC TAAATCATCT GTGCTTCTTC AATAGAGAAT CCTCAAACCA TTCAAACGAA 6240
AGTAGTAAAT TTTATTTTTT TATAATTTAT TTGATCAAGC TTTTAAGATT GCATTATTTT 6300
GAAGAGTAAA AAATAAAAAA ATATTAAATA AACTAGTTCG AAAATTCTAA CGTAATAAAA 6360
CTTCTCATTC GATTTGTGTT TGACTAATCA GTTTGTATCA TATGTATATT TTTCAGATTG 6420
ATAACTCAGG CTAAACACAA ACTGATTAGT CAAACATAGT ATACATATAA AAAGTCTAAC 6480
TATTGAGTCT AGTGGAGAGT TTTGGTGCTG GTGGAAAAAC ATGCATAACA TCGAGGGTGT 6540
ATCCAACGTA TCACCTCTCA AAACCACGAC CACCTTTTTG TACGTATTGT AGCTCCCACA 6600
TAGGTTGCAT AGCGATTCAT AATAATGCAC ATTTATTTGT CTTCAATAAT GGATCTGAGA 6660
CAATCACAAA TCGCTAAGTA TTATTACGTG TAAATAAACA GAAGTTATTA CCTAGACTCT 6720
GTTAGTGTTT TGAGACTCTT AATGCTTGGA GCATGGATGT ACCTAAGATG CACTAAATAT 6780
TTTTTTTTAA ACTCTGAGAA TTACGAACCT CGTACCTACA TGGATTCTAC GTGATTTATA 6840
AAAAAAAATA AAAGGAATTA TGTCTACAAC CATATATGTC TAAAGAGACA AAAATTGTGT 6900
CAAATTTAAT TTTCCTTAAT ACAGATGTTG GTATATACAG ATTTCTCTGT TTTTAACACA 6960
GTTTAAATTC AGTAGATGAT GTCCTCAAGA ATCCTCTATA ATTGTCTCTT AATTTATTTT 7020
GGTGAATTTG TCATCTACTA CAGGAGTTCT TAGGAGATAT TAACAGAGAA TTAAATAAAA 7080
CCACTTAAAA GAAGGCAAAG AGTGTGTATA TGGCTTGTCT AGTACCATAT ATATACTAAG 7140
AAAGAAATTT CTTCCGTTTC TCACACATAT ACCGAACAGA TCATGGTATA TATATGATTC 7200
TTTCTTTAAT GTTAGCTTTC CTTTTTGTTT TGTTACACAA TCAAATGTGT GGTCTTATGT 7260
AGAACTAATA CAATCGAAAG GAAAAACAAA ACAATGTGTT AGTTTACACA CCAGAATACA 7320
TCTTGATTAA TTTGGTAATA TTAGGCAAGT TGTTATGTGA CTTATTTTAT TCAAATATAA 7380
TAAGAAGTTT AAACCATTAT AATCCGTTCA ACAATACACT GAATAAAATA AGTTTATATT 7440
ATTCTTCAAC AAAGAGAAGA GTACAAGTAA GTAGAGGCGA ATTCAGGATT TGAAGAGTTT 7500
GGGTGCATCG TTTCTCTTCT CATGTTCATT CATCTCCGCT TAAGTCCTAA ACTTCTCAAA 7560
CCCACGTAGA ATATTATTTT AATTTAAGCA TTAGTAAAAA TATTAATGAC GGCTAACGAA 7620
TAATGTCATT TATAATAAAA TTAAATTCGT AATCATTTTT ATAATTACTG CCGATTGCTT 7680
ATTACAGTAA TTTAATAGCA AATAAACTCT TTAACTTGTA ATAAAATGAG TAAAAAATGG 7740
GTTGTTGGCT AAATTATCGT TTATTTGAGA AATTGAACAT TATTTTACTC ATTTTTTACC 7800
CAACAACCGC GAAATCGACT CTAGGTCGTA GGGGTGGACT CCCTTACTTC CAACAACGTA 7860
CTAGATCAAG CTTTAGCTGA GATCCAGCAT CCCCACCTGA GGGAATGAAG GTTGTTGCAT 7920
GATCTAGTTT GCCTTCTAAA GGTGCACTCT TTTTTATATT CTCTATTTTT ACTAGTTTCT 7980
CAAGATACTA CGGAAGATTT CCACGTGAGA AAAAATATAA GAGATAAAAA TGATCAAAGA 8040
GTTCTATGAT GTAGATCCGC CTCTGCAAGT AAGCGGAAGG ACAAATTTTT ACCTTACTAA 8100
CTCTATTGAA CATCTAGGCG GAGACGTTCA TTCGCCTTCC TGTTTAAAAA TGGAATGATT 8160
GAGATAACTG GCACCAAATC CCTATTAACA AGCATGAAAA TATGAGAAGA CGAAAGAACT 8220
AAGTAAGTTC CGTGGTTTAG GGATAATTGT TCGTACTTTT ATACTCTTCT GCTTTCTTGA 8280
TTCATTCAAA TAATTATTGT ATAATTTATA ACACACTCTA GGATAATATT ACAAATAAGA 8340
ATAGTGAATT ATTAATAACA TATTAAATAT TGTGTGAGAT CCTATTATAA TGTTTATTCT 8400
TATCACTTAA CTTAATTAAT GACGAACTAT AAAAGCAAAG AAGGAAGAGG AGTGATGAGC 8460
CTCGGAAAAT GAATTAATTA CTGCTTGATA TTTTCGTTTC TTCCTTCTCC TCACTACTCG 8520
GAGCCTTTTA AAAAATCACA AATGAATAAA GATGGCTAAA ATAGAGTAAC TTGATGAAGT 8580
TGATACTTCT TTTTTAGTGT TTACTTATTT CTACCGATTT TATCTCATTG AACTACTTCA 8640
ACTATGAAGC ATTCATAATC AAATCTGTTC AAAAACACTT GATAGTTAAT TTTTAATTTT 8700
AAGAGATGCG TAAGTATTAG TTTAGACAAG TTTTTGTGAA CTATCAATTA AAAATTAAAA 8760
TTCTCTACGA CAATATCGTT TCTTACTATT CTTTTAGGGC TTTTCGCCCT AGGGATATAG 8820
TATATATGAT GTTATAGCAA AGAATGATAA GAAAATCCCG AAAAGCGGGA TCCCTATATC 8880
ATATATACTC CAAATTTCAC GCCATATATA AATAATGTGG AAAAAACTTA TAATTTTCAA 8940
GGTAACAAGG GTTTAAAGTG CGGTATATAT TTATTACACC TTTTTTGAAT ATTAAAAGTT 9000
CCATTGTTCA AATCATATTT TTGTTACGCC TTCGTGGAGA CTACTTCCTC AATCGTAACA 9060
AAGAACAACT TTAGTATAAA AACAATGCGG AAGCACCTCT GATGAAGGAG TTAGCATTGT 9120
TTCTTGTTGA TTTCAAGTGG CGACTCAAAA AAGTCGCCAC GACCGGTGAT AACCTTTTGT 9180
GGCGATTATT AAAGTTCACC GCTGAGTTTT TTCAGCGGTG CTGGCCACTA TTGGAAAACA 9240
CCGCTAATAA TTTAGTCGCC ACTAAACATT ATTTGTGATC ATATTTTCTT TTCTTTTTCA 9300
TTTTTTATTT AAATCAGCGG TGATTTGTAA TAAACACTAG TATAAAAGAA AAGAAAAAGT 9360
AAAAAATAAT AAGGTCAAAT TTACCCCTTT ATCTTATTTA TGTAAATTGG AAAATCCCAA 9420
ATTTTGCATA TTCCAGTTTA AATGGGGAAA TAGAATAAAT ACATTTAACC TTTTAGGGTT 9480
TAAAACGTAT ATTTTTCGAA TTCCTTTTTT TTTAACACAC TCAAAAGTCA AAACATTAAA 9540
GAACGGAATA TAAAAAGCTT AAGGAAAAAA AAATTGTGTG AGTTTTCAGT TTTGTAATTT 9600
CTTGCCTTAA GCAAATTAAA TGGCAAAAGA CTTGTTCTAA CAAAAAATTA TTAGTAAAAC 9660
AGACTCATGT CGTTTAATTT ACCGTTTTCT GAACAAGATT GTTTTTTAAT AATCATTTTG 9720
TCTGAGTACA AATGTAACAA TAACCAACAA ATTATCCAAA ATTTTAGATA AATATTATCC 9780
AAACAAATAT TTACATTGTT ATTGGTTGTT TAATAGGTTT TAAAATCTAT TTATAATAGG 9840
TTTGTTTATA AAAATAATCA TCCAATCCAT TTAATTTATT TTTAAAAAAA ATCCTAAATT 9900
AACTCTCCAT TTTTATTAGT AGGTTAGGTA AATTAAATAA AAATTTTTTT TAGGATTTAA 9960
TTGAGAGGTT CTTTCACTCA AAAACAAACT CTACCCATTT TTTTCACTAT AAATACTCTT 10020
CATAATTTTA GAAAGTGAGT TTTTGTTTGA GATGGGTAAA AAAAGTGATA TTTATGAGAA 10080
GTATTAAAAC ATTTATTCTT CATTCCCAAG TTTCTTTCTC CCTATCCAAA AAAAATAAAA 10140
AAAAATTATG TAAATAAGAA GTAAGGGTTC AAAGAAAGAG GGATAGGTTT TTTTTATTTT 10200
TTTTTAATAT TAGATTAATT ACCACTATCT GTCAAAGCCC AATTATTGAA AAAAAAAAAA 10260
CTATGGATTA ATCTAATTAA TGGTGATAGA CAGTTTCGGG TTAATAACTT TTTTTTTTTT 10320
GATACCTAAA TTCATCTAAT TCTCGTTGGG CTTTGCCAGT TATCTTAGTT TGCTTTTTTA 10380
TAGTTTTATT AAGTAGATTA AGAGCAACCC GAAACGGTCA ATAGAATCAA ACGAAAAAAT 10440
ATCAAAATAT ATCCAATAAT GTTGTTTTTG CTTCTCACAA AGTTTTTATT CATTTGCAAT 10500
CTCAAAATGA TAGGTTATTA CAACAAAAAC GAAGAGTGTT TCAAAAATAA GTAAACGTTA 10560
GAGTTTTACC TGTAAATGTT CAAACTGTTC ATAGAACTGG TTACCATTTT CAGCCAGAAA 10620
AACATTGGAG ACATTTACAA GTTTGACAAG TATCTTGACC AATGGTAAAA GTCGGTCTTT 10680
TTGTAACCTT CAATGGTATG TATACGCGCT TTTTCGCGTT TTTTCTTCTT TATATATGTG 10740
TCAAAAAAAA GTTACCATAC ATATGCGCGA AAAAGCGCAA AAAAGAAGAA ATATATACAC 10800
AGTTTTTTTT A 10811
(2) INFORMATION FOR SEQ ID NO: 2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: other nucleic acid
(A) DESCRIPTION: /desc = "Forward primer"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: AACGATCCAA ATGGACCA 18
(2) INFORMATION FOR SEQ ID NO: 3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: other nucleic acid
(A) DESCRIPTION: /desc = "Reverse primer"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: GAAAAAATCA GGACATTCCC A 21
Claims
1. A pollen cell-specific expression control polynucleotide of an invertase gene, or a derivative, functional equivalent or part of said expression control polynucleotide.
2. An expression control polynucleotide as claimed in Claim 1 which is a promoter.
3. An expression control polynucleotide as claimed in either one of Claims 1 and 2 which comprises a sequence substantially as set out in SEQ ID No 1 or as present in NCTC Deposit No 13013, or a functional equivalent or part thereof .
4. An expression control polynucleotide as claimed in Claim 3 which comprises the sequence of nucleotides 3430-5349 of SEQ ID No 1.
5. A recombinant expression control polynucleotide comprising at least a part of a polynucleotide as claimed in any one of Claims 1 to 4.
6. A recombinant nucleotide construct which comprises an expression control polynucleotide as claimed in any one of Claims 1 to 5 operably linked to a heterologous polynucleotide.
7. A construct as claimed in Claim 6 which is in the form of a vector.
8. A construct as claimed in either one of Claims 6 and 7 wherein said heterologous polynucleotide encodes a protein.
9. A host cell transformed with a construct as claimed in any one of Claims 6 to 8.
10. A transgenic organism transformed with a construct as claimed in any one of Claims 6 to 8.
11. An organism as claimed in Claim 10 which is a plant .
12. An organism as claimed in Claim 11 wherein said expression control polynucleotide is pollen cell- specific and the heterologous polynucleotide operably linked thereto encodes for a protein which causes male sterility of said plant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU65110/98A AU6511098A (en) | 1997-03-19 | 1998-03-19 | Expression control polynucleotides |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB9705694.9 | 1997-03-19 | ||
| GBGB9705694.9A GB9705694D0 (en) | 1997-03-19 | 1997-03-19 | Polynucleotide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO1998041643A1 true WO1998041643A1 (en) | 1998-09-24 |
Family
ID=10809512
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/GB1998/000833 WO1998041643A1 (en) | 1997-03-19 | 1998-03-19 | Expression control polynucleotides |
Country Status (3)
| Country | Link |
|---|---|
| AU (1) | AU6511098A (en) |
| GB (1) | GB9705694D0 (en) |
| WO (1) | WO1998041643A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000023597A2 (en) * | 1998-10-16 | 2000-04-27 | Scottish Crop Research Institute | Tissue-specific promoters for gene expression |
| WO2000077187A2 (en) * | 1999-06-12 | 2000-12-21 | Thomas Roitsch | Promoter system and production and use of the same |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994001572A2 (en) * | 1992-07-09 | 1994-01-20 | Pioneer Hi-Bred International, Inc. | Maize pollen-specific polygalacturonase gene |
-
1997
- 1997-03-19 GB GBGB9705694.9A patent/GB9705694D0/en active Pending
-
1998
- 1998-03-19 AU AU65110/98A patent/AU6511098A/en not_active Abandoned
- 1998-03-19 WO PCT/GB1998/000833 patent/WO1998041643A1/en active Application Filing
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994001572A2 (en) * | 1992-07-09 | 1994-01-20 | Pioneer Hi-Bred International, Inc. | Maize pollen-specific polygalacturonase gene |
Non-Patent Citations (10)
| Title |
|---|
| ALLEN, R.L., ET AL.: "Molecular characterization of one of the maize polygalacturonase gene family members which are expressed during late pollen development", THE PLANT JOURNAL, vol. 3, no. 2, 1993, pages 261 - 271, XP002071297 * |
| HEDLEY, P.E., ET AL,: "cDNA cloning and expression of a potato (Solanum tuberosum) invertase", PLANT MOLECULAR BIOLOGY, vol. 22, 1993, pages 917 - 922, XP002071294 * |
| HEDLEY, P.E., ET AL.: "Potato (Solanum tuberosum) invertase-encoding cDNAs and their differential expression", GENE, vol. 145, 1994, pages 211 - 214, XP002071296 * |
| LORENZ, K., ET AL.: "D.carota (Queen Anne's Lace) Inv*Dc2 gene 3432bp", EMBL SEQUENCE DATABASE, ACCESSION NO. X78424, 25 March 1994 (1994-03-25) * |
| LORENZ, K., ET AL.: "Structural organization and differential expression of carrot beta-fructofuranosidase genes: identification of a gene coding for a flower bud-specific isozyme", PLANT MOLECULAR BIOLOGY, vol. 28, 1995, pages 189 - 194, XP002071295 * |
| MASCARENHAS, J.P.: "Gene activity during pollen development", ANN. REV. PLANT PHYSIOL. PLANT MOL. BIOL, vol. 41, 1990, pages 317 - 338, XP002071292 * |
| MEYER R ET AL: "Promoter deletion analysis of potato invertase gene expression.", ANNUAL MEETING OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY, SWANSEA, WALES, UK, APRIL 11-15, 1994. JOURNAL OF EXPERIMENTAL BOTANY 45 (SUPPL.). 1994. 6. ISSN: 0022-0957, XP002071293 * |
| RAMLOCH-LORENZ, K., E AL.: "Molecular characterization of the gene for carrot cell wall beta-fructosidase", THE PLANT JOURNAL, vol. 4, no. 3, 1993, pages 545 - 554, XP002071298 * |
| STURM A.: "D.carota (Queen Anne's Lace) Inv*Dc1 gene", EMBL SEQUENCE DATABASE, ACCESSION NO. X69321, 23 November 1992 (1992-11-23) * |
| XU J ET AL: "A similar dichotomy of sugar modulation and developmental expression affects both paths of sucrose metabolism: Evidence from a maize invertase gene family.", PLANT CELL 8 (7). 1996. 1209-1220. ISSN: 1040-4651, XP002071291 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000023597A2 (en) * | 1998-10-16 | 2000-04-27 | Scottish Crop Research Institute | Tissue-specific promoters for gene expression |
| WO2000023597A3 (en) * | 1998-10-16 | 2000-07-27 | Scottish Crop Research Inst | Tissue-specific promoters for gene expression |
| WO2000077187A2 (en) * | 1999-06-12 | 2000-12-21 | Thomas Roitsch | Promoter system and production and use of the same |
| WO2000077187A3 (en) * | 1999-06-12 | 2001-08-09 | Thomas Roitsch | Promoter system and production and use of the same |
| US7012173B1 (en) * | 1999-06-12 | 2006-03-14 | Freistaat Bayern | Promoter system and production and use of the same |
Also Published As
| Publication number | Publication date |
|---|---|
| AU6511098A (en) | 1998-10-12 |
| GB9705694D0 (en) | 1997-05-07 |
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