WO1997047647A1 - Proteines vegetales - Google Patents

Proteines vegetales Download PDF

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Publication number
WO1997047647A1
WO1997047647A1 PCT/ES1996/000130 ES9600130W WO9747647A1 WO 1997047647 A1 WO1997047647 A1 WO 1997047647A1 ES 9600130 W ES9600130 W ES 9600130W WO 9747647 A1 WO9747647 A1 WO 9747647A1
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WIPO (PCT)
Prior art keywords
protein
ser
leu
plant
nucleic acid
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PCT/ES1996/000130
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English (en)
Spanish (es)
Inventor
Crisanto Gutierrez-Armenta
Qi Xie
Andrés PELAYO SANZ-BURGOS
Paula Suarez Lopez
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Consejo Superior De Investigaciones Cientificas
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Priority to PCT/ES1996/000130 priority Critical patent/WO1997047647A1/fr
Priority to CA002257972A priority patent/CA2257972A1/fr
Priority to NZ333100A priority patent/NZ333100A/xx
Priority to AU32579/97A priority patent/AU721332B2/en
Priority to ZA975202A priority patent/ZA975202B/xx
Priority to EP97928187A priority patent/EP0914436A1/fr
Priority to CA002257828A priority patent/CA2257828A1/fr
Priority to PCT/EP1997/003070 priority patent/WO1997047745A1/fr
Priority to JP10501212A priority patent/JP2001502522A/ja
Priority to CN97197141A priority patent/CN1227605A/zh
Publication of WO1997047647A1 publication Critical patent/WO1997047647A1/fr
Priority to US09/213,293 priority patent/US6384299B1/en
Priority to US09/770,657 priority patent/US20020046416A1/en
Priority to US10/025,676 priority patent/US20020133847A1/en

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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)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
    • C12N15/8283—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for virus resistance
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • 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)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants

Definitions

  • the present invention relates to proteins that have biological activity in plant and animal systems, to polynucleotides that code for the expression of said proteins, to oligonucleotides for use in the identification and synthesis of these proteins and polynucleotides, to vectors and cells containing the Recombinant polynucleotides and plants and animals comprising these elements, and the use of proteins, polynucleotides and fragments thereof in the control of plant growth and the vulnerability of plants to viruses.
  • Rb retinoblastoma susceptibility gene
  • DNA tumor viruses that infect animal cells express oncoproteins that interact with the Rb protein through an LXCXE motif, breaking down Rb-E2F complexes and promoting the cells to enter the S phase (Weinberg ibid; Ludlow, J..W. PHASEB J. 7, 866 (1993); Moran, E. FASEB J. 7, 880 (1993); Vousden, K. FASEB J. 7, 872 (1993)).
  • the present inventors have shown that the efficient replication of a plant geminivirus requires the integrity of a motif of LXCXE amino acids in the RepA viral protein and that RepA can interact with members of the human Rb family in yeasts (Xie, Q. Suárez-López, P. and Gutiérrez, C. EMBO J. 14, 4073 (1995)).
  • the presence of the LXCXE motif has also been published in type D vegetable cyclines (Soni, R., Carmichael, JP, Shah, ZH and Murray, JAH Plant Cell 7, 85-103 (1995).
  • the present inventors have identified for the first time characteristic sequences of plant Rb proteins and the corresponding coding polynucleotides, they have isolated said proteins and polynucleotides and, in particular, have identified sequences that distinguish them from known animal Rb protein sequences.
  • the inventors have determined that a known DNA sequence from maize encoding a plant Rb protein and hereinafter referred to as ZmRbl.
  • ZmRbl a known DNA sequence from maize encoding a plant Rb protein and hereinafter referred to as ZmRbl.
  • the inventors have shown that ZmRbl interacts in yeasts with RepA, a vegetable geminivirus protein that contains an essential LXCXE motif for its function.
  • the inventors have further determined that the replication of geminivirus DNA in plant cells transfected with plasmids encoding human ZmRbl or pl30, a member of the human Rb family, is reduced.
  • Rb proteins from both humans and plants such as ZmRbl
  • ZmRbl have utility, among others, for therapeutic purposes for plants, diagnosis, growth control or research and many of these plant proteins will have similar utility in animals.
  • the use of retinoblastoma protein is provided in the growth control ' of plant cells and / or plant viruses.
  • the present invention provides control of viral infection and / or the growth of plant cells, where the virus requires the integrity of a motif of LXCXE amino acids in one of its proteins, particularly, for example, in the RepA viral protein. , for normal playback.
  • the particular plant viruses thus controlled are Geminivirus.
  • a preferred method of control using said proteins includes the application of these proteins to the plant cell, either directly or by introduction of DNA or RNA encoding for expression in the plant cell to be treated.
  • retinoblastoma protein By overexpression of the retinoblastoma protein or the expression of an Rb protein or a peptide fragment thereof that interacts with the virus's LXCXE motif, but does not affect the normal functioning of the cell, it is possible to inhibit viral growth. normal and, therefore, also reduce the spread of 'the infection of this cell to its neighbors.
  • antisense DNA or RNA into platelet cells in the form of vectors containing the promoters necessary for transcription of the DNA or RNA, it will be possible to exploit the well-known antisense mechanism to inhibit Rb protein expression, and by this of the S phase.
  • Such plants would be useful, among other aspects, to replicate DNA and RNA to high levels, for example in yeasts.
  • Methods for introducing antisense DNA into cells are well known to those skilled in the art: see for example "Principles of gene manipulation - An introduction to Genetic Engineering (1994) RW O ⁇ d & SB Primrose; Oxford-Blackwell Scientific Publications Fifth Edition p398 .
  • a recombinant nucleic acid is provided, particularly in the form of DNA or cRNA (mRNA), which codes for the expression of the Rb protein that is characteristic of plants.
  • This nucleic acid is characterized by one or several characteristic regions that differ from the nucleic acid of the known animal Rb protein and is exemplified herein by SEQ ID No 1, bases 31-2079.
  • the DNA or RNA may have a sequence containing degenerate substitutions in the nucleotides of the codons in SEQ ID No. 1, and where the RNA the T is U.
  • the most preferred DNA or RNA are capable of hybridizing with the polynucleotide of SEQ ID No. 1 under conditions of low stringency, preferably being able to hybridize under conditions of high stringency.
  • low stringency conditions and “high stringency conditions” are understood by the experts, but are conveniently used in US 5202257, Col-9-Col 10. If modifications were made that donate to the expression of a protein with different amino acids, preferably of the same class to the amino acids corresponding to SEQ ID No. 1; that is, they are conservative substitutions. Such substitutions are known to experts, for example, see US 5380712, and is contemplated only when the protein has activity with retinoblastoma protein.
  • the preferred DNA or cRNA encodes a plant Rb protein that has pocket subdomains A and B that have between 30% and 75% homology to the human Rb protein, particularly compared to pl30, more preferably between 50% and 64% homology.
  • the vegetable protein Rb thus encoded has conserved the amino acid C706 of human Rb.
  • the spacer sequence between the Pockets A and B are not conserved with respect to animal Rb proteins, preferably having a homology of less than 50% with the same region found in said animal proteins. More preferably, the protein thus encoded has a homology of 80% or greater with that of SEQ ID No 2 of the attached sequence list, even more preferably 90% or more and more preferably 95% or more.
  • a recombinant DNA of SEQ ID No. 1, bases 31 to 2079, or the entire SEQ ID No. 1, or the corresponding RNA encoding the corn cDNA clone encoding ZmRbl of SEQ ID No. 2 is provided.
  • the protein expressed by the recombinant DNA or RNA of the second aspect is provided, new proteins derived from said DNA or RNA and a protein derived from the naturally occurring DNA or RNA, by mutagenic means such as the use of mutagenic PCR primers.
  • vectors, cells, plants and animals comprising the recombinant DNA or RNA of correct or antisense sense of the invention are provided.
  • a method is provided to control cell or viral growth comprising the administration of DNA, RNA or proteins of the second or third aspect, to the cell.
  • Said administration may be direct in the case of proteins or may include indirect means, such as electroporation or plant seed cells with DNA or by transformation of cells with expression vectors capable of expressing or over-expressing the proteins of the invention or fragments of they are capable of inhibiting cell or viral growth.
  • the method uses an expression vector capable of producing antisense RNA from the cDNA of the invention.
  • plant protein and nuecleic acids includes an N-terminal domain corresponding in sequence to amino acids 1 to 90 of SEQ ID No. 2 and a nucleotide sequence corresponding to bases 31 to 300 of SEQ ID No. 1. These sequences are characterized by having less than 150 and less than 450 units than animal sequences that have more than 300 amino acids and 900 more base pairs.
  • Fig. 1 Sub-figure A shows the relative lengths of the present ZmRbl protein and human retinoblastoma proteins.
  • Sub-figure B shows the alignment of the amino acid sequences of the Pocket A and Pocket B of the ZmRbl with those of Xenopus, chicken, mouse and three human proteins (Rb, pl07 and pl30).
  • Fig. 2 This figure is a map of the main characteristics of the WDV virus and the pWori vector derived from WDV and the positions of the deletions and mutations used to establish that the LXCXE motif is required for replication in plant cells.
  • EXAMPLE 1 Isolation of DNA and protein expression clones
  • pBluéscript SK- (pBS) phagemids from the positive clones were excised by in vivo cleavage with the ExAssist (Stratagene) phage according to protocols recommended by the manufacturer.
  • DNA sequencing was carried out using a SequenaseTM (USB) device. The 5 'end of mRNAs encoding p75ZmRbl was determined by RACE-PCR. Poly-A + was purified
  • the first chain was synthesized using the oligonucleotide DraI35 (5'-GATTTAAAATCAAGCTCC, nucleotide positions 113-96). After denaturation at 90 ° C for 3 minutes, the RNA was removed by RNase treatment, the cDNA was recovered and a tail was created at the 5 'position with terminal transferase and dATP. Then, a PCR fragment was amplified using the DraI35 primer and the linker-primer (50 bp) from the Stratagene cDNA synthesis kit.
  • One of the positive clones (pBS.Rbl) thus produced contained a ⁇ 4 kb insert which, according to the restriction analysis, extended to the 5 'and 3' ends of the region contained in the Expressed Tag Sequence used.
  • the Nucleotide sequence corresponding to the longest cDNA insert (3747 bp) is shown in SEQ ID No 1.
  • This ZmRbl cDNA contains a single open reading phase capable of encoding a 683 amino acid protein (predetermined Mr 75247, p75ZmRbl) followed for a 3 'untranslated region of 1646 bp. Non-translated regions of similar length have also been found in mammalian Rb cDNA (Lee, W.
  • Plasmid pWori ⁇ was constructed by deleting in pWori the majority of the sequences encoding WDV proteins (Sanz and Gutierrez, unpublished). Plasmid p35S.Rbl was constructed by inserting the CaMV 35S promoter (obtained from pWDV3: 35SGUS) upstream of the ZmRBl cDNA into the pBS vector. Plasmid p35S.130 was constructed by introducing the complete coding sequence of human pl30 instead of ZmRbl sequences within p35S.Rbl. Plasmid p35.A + B was constructed by replacing sequences encoding the ORFs of Rep A and Rep B of WDV instead of ZmRbl in plasmid p35S.Rbl
  • the ZmRbl protein contains segments homologous to subdomains A and B of the "pocket" that is present in all members of the Rb family. These subdomains are separated by an unconserved spacer.
  • the ZmRbl also contains non-conserved N-terminal and C-terminal domains.
  • ZmRbl shares an amino acid identity of -28-30% (similarity of -50%) with members of the Rb family (Hannon, GJ, Demetrick, D. & Beach, D. Genes Dev. 7, 2378 (1993); Cobrinik, D., Whyte, P., Peeper, DS, Jacks, T. & Weinberg, RA ibid., P. 2392 (1993).
  • Amino acids 561-577 encompass a proline-rich domain.
  • ZmRbl contains 16 consensus sites, SP or TP, for phosphorylation by cyclin-dependent kinases (CDKs) with one at the 5 'end of subdomain A and several in the C-terminal zone that are potential phosphorylation sites.
  • CDKs cyclin-dependent kinases
  • Dwarf wheat geminivirus (WDV) replication is dependent on an intact LXCXE motif of the viral RepA protein. This motive may mediate the interaction with a member of the human Rb family, pl30, in yeasts. Therefore, the inventors investigated whether p75ZmRbl could be complexed with WDV RepA using the two yeast hybrid system (Fields, S. and Song, O. Nature 340, 245-264 (1989)). Yeast cells were cotransformed with a plasmid encoding the GAL4BD-RepA fusion protein and with plasmids encoding different GAL4AD fusion proteins.
  • the GAL4AD-p75ZmRbl fusion could also be complexed with GAL4BD-RepA to allow the growth of recipient yeast cells in the absence of histidine. This interaction was slightly stronger than that observed with the human protein pl30.
  • the RepA was also able to join to some extent the truncated form in the N-terminal of p75ZmRbl.
  • the role of the LXCXE motif in the RepA-p75ZmRbl interaction was determined using a point mutation in WDV RepA
  • the E198K mutant of WDV RepA behaves similarly to point-like mutations of animal virus oncoproteins (Moran, E., Zerler, B., Harrison, TM and Mathews, MB Mol. Cell Biol. 6, 3470 (1986); Cherington, V. et al., Ibid, p. 1380 (1988); Lillic, JW, Lowenstein, PM, Green, MR and Green, M. Cell 50, 1091 (1987); DeCaprio, JA et al., Ibid., P.275 (1988)).
  • yeast cotransformants are related to the levels of b-galactosidase activity.
  • PCNA proliferating cell nuclear antigen
  • the accumulation of newly replicated viral plasmid DNA was impaired in wheat cells transfected with plasmids expressing human p75ZmRbl or pl30, when the expression of the WDV replication protein (s) is directed by the WDV promoter or by the CaMV 35S promoter.
  • p75ZmRbl could also play key regulatory roles at other levels during the life of the plant cell.
  • a key issue that arises from the existence of Rb homologs in plant cells is whether, as in animals, the disruption of the Rb pathway leads to a tumor tendency disorder.
  • the inventors have observed that the VirB4 protein encoded by the Ti plasmids of both AgroJbacterium tumefaciens and
  • VirB4 protein is necessary for tumor induction
  • wheat cells were co-transfected with pWori (0.5 g) and pBS plasmids (control), p35S.Rbl or p35S.130 (10 g in each case ). Replication of the test plasmid (p Wori) was analyzed two days after transfection and was detected as described in part A using ethidium bromide marking; and hybridization of Southern C.
  • the pWoriDD test plasmid (which does not encode replication proteins) of functional WDVs but replicates when they are provided by a different plasmid, i.e., pWori).
  • Wheat cells were co-transfected, as indicated, with pWori ⁇ (0.25 g), pWori (0.25 g) p35S.AB (6 g), p35S.Rbl (10 g) and / or p35S.130 (10 g).
  • Replication of the test plasmid (pWoriDD) was analyzed 36 hours after transfection and was detected as described in part A using ethidium bromide, Southern hybridization. Plasmids pWori (MI) and pWori ⁇ were used
  • ORGANISM Zea mays (ix) CHARACTERISTICS:
  • GAT TTT AAA TCA ATT TTG ATC AAT AAT GAT TAT ATT CCC TAT GAT GAG 150 Asp Phe Lys Ser lie Leu He Asn Asn Asp Tyr lie Pro Tyr Asp Glu 25 30 35 40
  • AGT ATC TAT ATT GGG AGT ACG AAC CGT AAT GGG GTA TTA GTA TCG CGC 1590 Ser He Tyr He Gly Ser Thr Asn Arg Asn Gly Val Leu Val Ser Arg 505 510 515 520
  • GACGCTGACTCATCTC ⁇ ATAATGGCAGATGCTTAACCATCTTTAGGAGCTCATGTCATGA3259

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Abstract

La présente invention concerne l'isolement et la caracterisation d'une séquence d'ADN de cellules de plantes qui code une protéine rétinoblastome. Cette découverte se base sur les propriétés structurelles et fonctionnelles de la protéine de rétinoblastome de plantes en tant que régulateur du cycle cellulaire, de la croissance cellulaire et de la différentiation cellulaire chez les plantes. A cet effet, on révendique, entre autres, l'utilisation de la protéine rétinoblastome ou la séquence d'ADN qui la code dans la régulation de la croissance de cellules végétales, de plantes et/ou de virus relatifs aux végétaux, ainsi que l'utilisation de vecteurs, cellules, plantes ou animaux ou cellules animales modifiés par manipulation du processus de régulation basé sur la protéine rétinoblastome de plantes.
PCT/ES1996/000130 1996-06-13 1996-06-13 Proteines vegetales WO1997047647A1 (fr)

Priority Applications (13)

Application Number Priority Date Filing Date Title
PCT/ES1996/000130 WO1997047647A1 (fr) 1996-06-13 1996-06-13 Proteines vegetales
CA002257972A CA2257972A1 (fr) 1996-06-13 1996-06-13 Proteines vegetales
EP97928187A EP0914436A1 (fr) 1996-06-13 1997-06-12 Proteines vegetales associees au retinoblastome
AU32579/97A AU721332B2 (en) 1996-06-13 1997-06-12 Plant retinoblastoma-associated proteins
ZA975202A ZA975202B (en) 1996-06-13 1997-06-12 Plants proteins
NZ333100A NZ333100A (en) 1996-06-13 1997-06-12 Plant retinoblastoma-associated proteins and their use in modulating the cell cycle of a plant
CA002257828A CA2257828A1 (fr) 1996-06-13 1997-06-12 Proteines vegetales associees au retinoblastome
PCT/EP1997/003070 WO1997047745A1 (fr) 1996-06-13 1997-06-12 Proteines vegetales associees au retinoblastome
JP10501212A JP2001502522A (ja) 1996-06-13 1997-06-12 網膜芽細胞腫関連植物蛋白
CN97197141A CN1227605A (zh) 1996-06-13 1997-06-12 植物成视网膜细胞瘤相关蛋白
US09/213,293 US6384299B1 (en) 1996-06-13 1998-12-14 Plant retinoblastoma-associated gene
US09/770,657 US20020046416A1 (en) 1996-06-13 2001-01-29 Plant proteins
US10/025,676 US20020133847A1 (en) 1996-06-13 2001-12-26 Plant proteins

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PCT/ES1996/000130 WO1997047647A1 (fr) 1996-06-13 1996-06-13 Proteines vegetales
CA002257972A CA2257972A1 (fr) 1996-06-13 1996-06-13 Proteines vegetales

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US21329498A Continuation 1996-06-13 1998-12-14

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PCT/EP1997/003070 WO1997047745A1 (fr) 1996-06-13 1997-06-12 Proteines vegetales associees au retinoblastome

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EP (1) EP0914436A1 (fr)
CN (1) CN1227605A (fr)
AU (1) AU721332B2 (fr)
CA (2) CA2257972A1 (fr)
WO (2) WO1997047647A1 (fr)
ZA (1) ZA975202B (fr)

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WO1998056811A2 (fr) * 1997-06-12 1998-12-17 Consejo Superior De Investigaciones Cientificas Proteines vegetales grab
WO2002074909A2 (fr) * 2001-03-16 2002-09-26 Pioneer Hi-Bred International, Inc. Acides nucleiques et polypeptides intervenant dans le cycle cellulaire, et leurs utilisations
US6696560B1 (en) 1999-03-19 2004-02-24 The United States Of America As Represented By The United States Department Of Energy Retinoblastoma-like RRB gene of arabidopsis thaliana

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WO1999066055A2 (fr) * 1998-06-15 1999-12-23 Cropdesign N.V. Sequences de regulation inductibles par des pathogenes de vegetaux, liees de maniere operationnelle a des genes du cycle cellulaire, et utilisation desdites sequences
US6284947B1 (en) * 1999-02-25 2001-09-04 Pioneer Hi-Bred International, Inc. Methods of using viral replicase polynucleotides and polypeptides
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THE EMBO JOURNAL, 1995, Vol. 14, No. 16, XIE Q. et al., "Identification and Analysis of a Retinoblastoma Binding Motif in the Replication Protein of a Plant DNA Virus: Requirement for Efficient Viral DNA Replication", pp. 4073-4082. *
THE PLANT CELL, January 1995, Vol. 7, SONI R. et al., "A Family of Cyclin D Homologs from Plants Differentially Controlled by Growth Regulators and Containing the Conserved Retinoblastoma Protein Interaction Motif", pages 85-103. *
THE PLANT CELL, November 1995, Vol. 7, DAHL M. et al., "The D-Type Alfalfa Cyclin Gene Cyc Ms 4 Complements G1 Cyclin-Deficient Yeast and is Induced in the G1 Phase of the Cell Cycle", pages 1847-1857. *
VIROLOGY, May 1996, Vol. 219, COLLIN S. et al., "The Two Nonstructural Proteins from Wheat Dwarf Virus Involved in Viral Gene Expression and Replication Are Retinoblastoma Binding Proteins", pages 324-329. *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998056811A2 (fr) * 1997-06-12 1998-12-17 Consejo Superior De Investigaciones Cientificas Proteines vegetales grab
WO1998056811A3 (fr) * 1997-06-12 1999-03-04 Consejo Superior Investigacion Proteines vegetales grab
US6696560B1 (en) 1999-03-19 2004-02-24 The United States Of America As Represented By The United States Department Of Energy Retinoblastoma-like RRB gene of arabidopsis thaliana
WO2002074909A2 (fr) * 2001-03-16 2002-09-26 Pioneer Hi-Bred International, Inc. Acides nucleiques et polypeptides intervenant dans le cycle cellulaire, et leurs utilisations
WO2002074909A3 (fr) * 2001-03-16 2003-03-13 Pioneer Hi Bred Int Acides nucleiques et polypeptides intervenant dans le cycle cellulaire, et leurs utilisations

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CA2257828A1 (fr) 1997-12-18
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WO1997047745A1 (fr) 1997-12-18
CA2257972A1 (fr) 1997-12-18
AU3257997A (en) 1998-01-07
ZA975202B (en) 1998-12-14
AU721332B2 (en) 2000-06-29

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