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Patent US5162601 - Plant potyvirus expression vector with a gene for protease - Google PatentsSearch Images Maps Play YouTube News Gmail Drive More »Sign inPatentsThe present invention relates to a recombinant multigene comprising a plurality of structural genes and a plurality of DNA sequences which encode peptide linkers. In the present invention, one of the structural genes encodes a protease, the DNA sequences encoding the peptide linkers are adjacent to the...http://www.google.com/patents/US5162601?utm_source=gb-gplus-sharePatent US5162601 - Plant potyvirus expression vector with a gene for proteaseAdvanced Patent SearchTry the new Google Patents, with machine-classified Google Scholar results, and Japanese and South Korean patents.Publication numberUS5162601 APublication typeGrantApplication numberUS 07/752,972Publication dateNov 10, 1992Filing dateAug 30, 1991Priority dateNov 22, 1989Fee statusPaidPublication number07752972, 752972, US 5162601 A, US 5162601A, US-A-5162601, US5162601 A, US5162601AInventorsJerry L. SlightomOriginal AssigneeThe Upjohn CompanyExport CitationBiBTeX, EndNote, RefManNon-Patent Citations (20), Referenced by (112), Classifications (27), Legal Events (11) External Links: USPTO, USPTO Assignment, EspacenetPlant potyvirus expression vector with a gene for protease
US 5162601 AAbstract
The present invention relates to a recombinant multigene comprising a plurality of structural genes and a plurality of DNA sequences which encode peptide linkers. In the present invention, one of the structural genes encodes a protease, the DNA sequences encoding the peptide linkers are adjacent to the DNA sequences which encode the structural genes and the peptide linkers contain an amino acid sequence which the protease recognizes as a proteolytic cleavage site. The present invention additionally relates to transgenic plants which contain such a recombinant multigene transgene, host cells transformed with such a recombinant multigene, and transgenic animals which contain such a recombinant multigene transgene. Furthermore, the present invention relates to a method of producing a plurality of polypeptides in a host by incorporating and expressing in the host a such recombinant multigene.
1. A recombinant multigene vector comprising: a) a plurality of structural genes; andb) a plurality of DNA sequences which encode peptide linkers; wherein at least one of said structural genes encodes a potyvirus protease; DNA sequences adjacent to said structural genes are said DNA sequences which encode peptide linkers; and said peptide linkers contain an amino acid sequence which said potyvirus protease recognizes as a protease cleavage site. 2. A recombinant multigene vector according to claim 1 wherein said potyvirus protease is selected from the group consisting of tobacco etch virus protease, watermelon mosaic virus 2 protease, zucchini yellow mosaic virus protease, and papaya ringspot 2 virus protease.
This application is a File Wrapper Continuation application of U.S. patent application Ser. No. 07/441,092 filed Nov. 22, 1989, abandoned.
The present invention relates to a recombinant DNA molecule useful in introducing genetic information into host organisms.
The techniques of genetic engineering allow for the introduction of specific genetic information into a host regardless of the source of such information. Accordingly, it is possible to add foreign derived information to a host's genetic material. Moreover, it is possible to add additional copies of endogenous genetic material to a host. In either case, if the additionally provided genetic information introduced into the host is expressed, the result of such expression is to confer and/or enhance a desired trait or traits.
Dougherty, W. G. et al., Virology, 172:302-310 (1989) report the effects of altering the amino acid sequence of the tobacco etch virus 49K protease on proteolysis by site directed mutagenesis of the protease cDNA gene. The findings disclosed suggest the catalytic triad of the protease to be composed of the His234, Asp269, and Cys339.
The present invention relates to a recombinant multigene comprising a plurality of structural genes and a plurality of DNA sequences which encode peptide linkers. In the present invention, one of the structural genes encodes a protease, the DNA sequences encoding the peptide linkers are adjacent to the DNA sequences which encode the structural genes and the peptide linkers contain an amino acid sequence which the protease recognizes as a proteolytic cleavage site.
Certain conventions are used in Charts 1-7 to illustrate plasmids and DNA fragments as follows:
Pca =CaMV 35S Promoter;
Icm =CMV intergenic region including 5' untranslated region and imitation codon according to Kozak's element;
Sca =CaMV polyadenylation signal;
Protev =TEV protease gene;
TEVcp =TEV coat protein gene;
PRVcp =PRV coat protein gene;
Psv =SV40 promoter;
HtPA =Human tPA gene;
StPA =Human tPA polyadenylation signal.
PL =λ phage PL promoter
Construction of an expressible Multigene containing the Tobacco Etch Virus (TEV) protease genes and two potyvirus coat protein genes (TEV and PRV)
This example describes a method of assembling regulatory elements and known genes to produce an expressible multigene Polymerase Chain Reaction (PCR) technology is used in order to produce copies of the desired genetic sequences. Furthermore, desired restriction enzyme sites are added at both the 5' and 3' end of copies made. The techniques used are referred to as Custom Polymerase Chain Reaction (CPCR) to describe the PCR technique used to amplify genetic sequences and add custom restriction endonuclease sites at the 5' and 3' ends of such sequences.
As shown in Chart 2, the desired region from pUC18cp-exp includes the promoter from CaMV (Pca); the intergenic region from CMV (Icm) which includes the 5'-untranslated region and the initiation codon according to Kozak's Element; and the 3'-untranslated region including the polyadenylation signal from CaMV (Sca). Using CPCR technology, oligomers were produced which amplify the expression cassette in pUC18-cp-exp and add XbaI restriction sites on the 5'-end of the Pca and 3'-end of Sca. Accordingly, the desired portion of pUC18cp-exp may be produced having XbaI sites at both the 5' and 3' ends. The product of this reaction is referred to as cp-exp fragment. It may be treated with XbaI and cloned into the XbaI site of pUCl8. The resulting plasmid is referred to as P18Xcp-exp and is shown in Chart 2. The following oligomer sequences can be used for this amplification: ##STR1##
Seeds of Phaseolus vulgaris cv. Olathe or Glycine max (cV.A0949) are surface sterilized with 15% Clorox for 10 minutes, followed by 4-5 rinses with distilled water and then placed on moistened paper towels in a temperature controlled Percival incubator at 28� C. and allowed to germinate for various times, 16 to 96 hours. Seed coats are removed and the decoated seeds are opened in halves (that is how cotyledons were removed from the main seed body). The mesocotyl region of the germinating seeds, with their plummule still attached, are infected with an overnight liquid culture of various Agrobacterium strains by using an Eppendorf pipetter fitted with a 271/2 gauge needle. Seeds are infected with virulent or non-virulent A. tumefaciens strains (A208C58, C58z707 and A208/phas-zein) or A. rhizogenes strains (A4RS and A4RS(pR:B278b)pu3.3c-1]. The common A. tumefaciens and A. rhizogenes strains are available from ATCC, 12301 Parklawn Drive, Rockville, MD. The disarmed A. rhizogenes strain RS(pRiB278b) has been described by Vilaine and Casse-Debart (1987) Mol. Gen. Genet., 206,17 and is available from Dr. F. Casse-Delbart, C.N.R.A., Routede Saint Cyr, F78000, Versailles, France. The disarmed A. tumefaciens, strain C582707 is available from Dr. A. G. Hepburn, University of Illinois, Urbana, IL. Inoculated seeds are then placed on moistened paper towels in petri dishes and incubated at 28� C. After four days these seedlings are transformed to soil and grown to maturity in the greenhouse. Plants infected with virulent strains of A. tumefaciens are scored for efficiency of gall formation as a function of germination time.
NPT II enzyme activity is detected by a standard in situ gel assay. Briefly, 100 mg. of a leaf tissue was mixed with 20 ml. of extraction buffer in a 1.5 ml. Eppendorf tube. Tissue samples are macerated with a Konte pestle and centrifuged for 20 minutes at 4� C. A 35 μl aliquot of the supernatant solutions was electrophoresed on a non-denaturing 10% polyacrylamide gel. The gel was overlaid with a 1% agarose gel containing 67 mM. tris-maleate (pH 7.1), 42 mM. MgCl2, 400 mM NH4 Cl, 20 μg kanamycin sulfate and 200 μCi γ-[32 P]ATP. After incubating for 30 minutes at room temperature, the agarose gel is blotted onto Whatman P81 phosphocellulose paper overnight. The P81 paper is removed, washed several times with hot water (80� C.) and autoradiographed.
Construction of a Multigene containing the tobacco etch virus protease gene, the human renin gene, and the human tPA gene under the control of the SV40 promoter.
This example describes a multigene which has the TEV protease and two human structural genes under the control of Simian Virus 40 (SV40) promoter. Using the SV40 promoter, multigenes of this construction may be used for expression in animal systems, either cellular or in transgenic animals. The SV40 promoter is widely used in animal expression systems (Moreau et al, 1981 Nuc. Acids Res. 9:6047-68). The multigene construction described in this example is useful for the expression of cDNA clones of both human renin (Imai et al, 1983 Proc. Natl. Acad. Sci. USA 80:7405-9) and human tissue plasminogen activator (tPA) (Pennica et al, Nature 301:214-21).
The first step in this construction is to use CPCR technology to amplify SV40 promoter and add appropriate restriction enzyme sites at the 5' and 3' ends (see Chart 4). Selection of restriction enzyme sites can be adjusted to avoid sites internal to any of the particular genes which are to be used in the multigene. The copies are generated from wildtype SV40 DNA. The 5' oligomer used to amplify the SV40 promoter adds an EcoRI site to the 5' end. The 3' oligomer used in the amplification adds an NcoI site at the initiation codon at the 3' end. The resulting fragment, referred to as Psv fragment in Chart 4 contains a copy of the SV40 promoter with an EcoRI site at its 5' end and an EcoRI site and a NcoI site at the 3' end downstream from the initiation codon. The following oligomer primers are used to generate Psv fragment: ##STR5##
The amplified SV40 promoter (400 bp) and TEV protease (1300 bp) fragments are then treated with NcoI and ligated to each other. The resulting 1700 bp fragment, referred to as Psv /TEV fragment in Chart 4, is treated with EcoRI and inserted into the plasmid pUC18 which is opened with EcoRI. The resulting recombinant plasmid is referred to as pUC18SVTEV and shown in Chart 4. This plasmid contains an inframe KpnI site in the SVTEV insert which can be used to clone additional genes.
Multigene construct for expression in prokaryotes
The following example illustrates a construction which can be used to express in a bacterial expression system, such as E. coli a multigene which contains a protease. This construction follows directly from Example 2 which describes the major elements needed except that instead of using the SV40 promoter, a bacterial promoter is used. In this example the λ phage PL promoter is used. The PL promoter can be obtained from any number of E. coli expression vectors or from the λ phage DNA. In this example, an EcoRI site to the 5'-end and an NcoI site to the 3'-end of the promoter by CPCR technology to form PL fragment as shown in Chart 8. The following oligomer primers are used: ##STR9##
After CPCR amplification, the 120 bp PL promoter fragment is subjected to digestion with NcoI and ligated with the 1 kb TEV protease gene containing fragment referred to as TEV protease fragment described in Example 2. The product of this ligation, referred to as PL TEV fragment in Chart 8, is then digested with EcoRI and cloned into pUC18 to obtain the plasmid referred to as pUC18PL TEV in Chart 8. As described in Example 2, the TEV protease fragment which is incorporated in the vector pUC18PL TEV contains an inframe KpnI site which can be used as a cloning site. The multigene construction RenPSJtPA fragment which was described in Example 2 and shown in Chart 7 can be inserted in the KpnI site of pUC18PL TEV. The clone resulting from the addition of the PL promoter to the human renin and tPA genes is referred to as pUC18PL TEVRentPA and shown in Chart 8.
A host bacteria can be transformed with plasmid pUC18PL TEVRentPA and the multigene will be expressed. The translation product will be processed by the TEV protein and both human renin and human tPA will be produced by the transformed bacteria.
Expression of a multigene in a eukaryotic host
Plasmid pSVTEVRentPA, described in Example 2, is digested with EcoR1 and the fragment containing the multigene under the regulatory control of the SV40 promoter is isolated. Chinese Hamster Ovary (CHO) cells are transfected with the multigene fragment. Cells are selected which express human renin and human tPA. The techniques used are widely known to those having ordinary skill in the art.
Production of a transgenic animal using a recombinant multigene as the transgene
Plasmid pSVTEVRentPA, described in Example 2, is digested with EcoR1 and the fragment containing the multigene under the regulatory control of the SV40 promoter is isolated. The recombinant multigene is introduced into non-human animal embryos using the methods described in Wagner, T.E. et al, Microinjection of a rabbit β-globin gene into zygotes and its subsequent expression in adult mice and their offspring. Proc. Natl. Acad. Sci. USA Vol. 78, No. 10 pp. 6376-6380, (Oct. 1981). Between 200 and 400 copies of the recombinant multigene are microinjected into each male pronuclei. Several lines of transgenic animals which express both human renin and human tPA are generated. The copy number for the multigene construct range between 5 to 10 copies per genome. ##STR10##
Non-Patent CitationsReference1Allison, R., et al., "The Nucleotide Sequence of the Coding Region of Tobacco Etch Virus Genomic RNA: Evidence for the synthesis of a Single Polyprotein", 1986, Virology 154:9-20.2 *Allison, R., et al., The Nucleotide Sequence of the Coding Region of Tobacco Etch Virus Genomic RNA: Evidence for the synthesis of a Single Polyprotein , 1986, Virology 154:9 20.3 *Carrington et al. Proced. Nat l Acad. Sci. 1988 vol. 85, 3391 3395.4Carrington et al. Proced. Nat'l Acad. Sci. 1988 vol. 85, 3391-3395.5Carrington, J. C., et al., "Small Nuclear Inclusion Protein Encoded by a Plant Potyvirus Genome is a Protease", 1987, J. Virology 61:2540-2548.6 *Carrington, J. C., et al., Small Nuclear Inclusion Protein Encoded by a Plant Potyvirus Genome is a Protease , 1987, J. Virology 61:2540 2548.7Dougherty, W. G., et al., "Biochemical and Mutational Analysis of Plant Virus Polyprotein Cleavage Site", 1988, Embo J. 7:1281-1287.8Dougherty, W. G., et al., "Characterization of the Catalytic Residues of the Tobacco Etch Virus 49-kDa Proteinase", 1989, Virology 172:302-310.9Dougherty, W. G., et al., "Expression and Function of Potyviral Gene Products", 1988, Ann. Rev. Phytopathol. 26:123-143.10Dougherty, W. G., et al., "Molecular Genetic and Biochemical Evidence for the Involvement of the Heptapeptide Cleavage Sequence in Determining the Reaction Profile at Two Tobacco Etch Virus Cleavage Sites in Cell-Free Assays", 1989, Virology 172:145-155.11Dougherty, W. G., et al., "Nucleotide Sequence at the 3' Terminus of Pepper Mottle Virus Genomic RNA: Evidence for an Alternative Mode of Potyvirus Capsid Protein Gene Organization", 1985, Virology 146:282-291.12 *Dougherty, W. G., et al., Biochemical and Mutational Analysis of Plant Virus Polyprotein Cleavage Site , 1988, Embo J. 7:1281 1287.13 *Dougherty, W. G., et al., Characterization of the Catalytic Residues of the Tobacco Etch Virus 49 kDa Proteinase , 1989, Virology 172:302 310.14 *Dougherty, W. G., et al., Expression and Function of Potyviral Gene Products , 1988, Ann. Rev. Phytopathol. 26:123 143.15 *Dougherty, W. G., et al., Molecular Genetic and Biochemical Evidence for the Involvement of the Heptapeptide Cleavage Sequence in Determining the Reaction Profile at Two Tobacco Etch Virus Cleavage Sites in Cell Free Assays , 1989, Virology 172:145 155.16 *Dougherty, W. G., et al., Nucleotide Sequence at the 3 Terminus of Pepper Mottle Virus Genomic RNA: Evidence for an Alternative Mode of Potyvirus Capsid Protein Gene Organization , 1985, Virology 146:282 291.17Dougherty; W. G., et al., "Molecular Genetic Analysis of a Plant Virus Polyprotein Cleavage Site: A Model", 1989, Virology 171:356-364.18 *Dougherty; W. G., et al., Molecular Genetic Analysis of a Plant Virus Polyprotein Cleavage Site: A Model , 1989, Virology 171:356 364.19Yeh, S. D., et al., "Translation of Papaya Ringspot virus RNA in Vitro: Detection of a Possible Polyprotein That is Processed for Capsid Protein, Cylindrical-Inclusion Protein and Amorphous-Inclusion Protein", 1985, Virology 143:260-271.20 *Yeh, S. D., et al., Translation of Papaya Ringspot virus RNA in Vitro: Detection of a Possible Polyprotein That is Processed for Capsid Protein, Cylindrical Inclusion Protein and Amorphous Inclusion Protein , 1985, Virology 143:260 271.* Cited by examinerReferenced byCiting PatentFiling datePublication dateApplicantTitleUS5491076 *Nov 1, 1993Feb 13, 1996The Texas A&M University SystemExpression of foreign genes using a replicating polyprotein producing virus vectorUS5530187 *Jul 16, 1993Jun 25, 1996The Salk Institute For Biological StudiesTransgenic plants containing multiple disease resistance genesUS5532142 *Feb 12, 1993Jul 2, 1996Board Of Regents, The University Of Texas SystemMethod of isolation and purification of fusion polypeptidesUS5766885 *Jun 6, 1995Jun 16, 1998Texas A & M UniversityPotyvirus vectors for the expression of foreign genesUS5846767 *Dec 19, 1994Dec 8, 1998Zeneca LimitedExpression of self processing polyproteins in transgenic plantsUS5912167 *Jun 6, 1995Jun 15, 1999Wisconsin Alumni Research FoundationAutocatalytic cleavage site and use thereof in a protein expression vectorUS5955072 *Jul 11, 1995Sep 21, 1999Sankyo Company, LimitedExpression systems utilizing autolyzing fusion proteins and a reducing polypeptideUS5965124 *Jan 31, 1995Oct 12, 1999Whitehead Institute For Biomedical ResearchReplication-competent recombinant viral vaccines and method of producing sameUS6002072 *Jun 7, 1995Dec 14, 1999Seminis Vegetable Seeds, Inc.Coat protein gene for the FLA83 W strain of papaya ringspot virusUS6150588 *Apr 4, 1997Nov 21, 2000Zeneca LimitedDNA encoding antimicrobial proteins from impatiensUS6307038 *Oct 6, 1998Oct 23, 2001Sankyo Company, LimitedExpression systems utilizing autolyzing fusion proteins and a novel reducing polypeptideUS6503732Sep 21, 1999Jan 7, 2003The Scripps Research InstituteMethod for using tobacco mosaic virus to overproduce peptides and proteinsUS6743611Apr 25, 2001Jun 1, 2004Sankyo Company, LimitedExpression systems utilizing autolyzing fusion proteins and a novel reducing polypeptideUS7091401Dec 20, 2002Aug 15, 2006Sembiosys Genetics Inc.Expression of epidermal growth factor in plant seedsUS7632823Dec 15, 2009Ambrx, Inc.Compositions of tRNA and uses thereofUS7632924Dec 15, 2009Ambrx, Inc.Antigen-binding polypeptides and their usesUS7638299Dec 29, 2009Ambrx, Inc.Biosynthetic polypeptides utilizing non-naturally encoded amino acidsUS7736872Dec 1, 2005Jun 15, 2010Ambrx, Inc.Compositions of aminoacyl-TRNA synthetase and uses thereofUS7816320Oct 19, 2010Ambrx, Inc.Formulations of human growth hormone comprising a non-naturally encoded amino acid at position 35US7829310Nov 9, 2010Ambrx, Inc.Compositions of aminoacyl-tRNA synthetase and uses thereofUS7838265Mar 15, 2010Nov 23, 2010Ambrx, Inc.Compositions of aminoacyl-tRNA synthetase and uses thereofUS7846689Mar 15, 2010Dec 7, 2010Ambrx, Inc.Compositions of aminoacyl-tRNA synthetase and uses thereofUS7858344Mar 15, 2010Dec 28, 2010Ambrx, Inc.Compositions of aminoacyl-tRNA synthetase and uses thereofUS7883866Feb 8, 2011Ambrx, Inc.Compositions of aminoacyl-tRNA synthetase and uses thereofUS7919591Apr 5, 2011Ambrx, Inc.Modified human plasma polypeptide or Fc scaffolds and their usesUS7939496Dec 21, 2005May 10, 2011Ambrx, Inc.Modified human growth horomone polypeptides and their usesUS7947473May 24, 2011Ambrx, Inc.Methods for expression and purification of pegylated recombinant human growth hormone containing a non-naturally encoded keto amino acidUS7959926Jun 14, 2011Ambrx, Inc.Methods for expression and purification of recombinant human growth hormone mutantsUS8012931Mar 19, 2008Sep 6, 2011Ambrx, Inc.Modified FGF-21 polypeptides and their usesUS8022186Sep 20, 2011Ambrx, Inc.Modified human plasma polypeptide or Fc scaffolds and their usesUS8053560Nov 8, 2011Ambrx, Inc.Modified human plasma polypeptide or Fc scaffolds and their usesUS8080391Dec 21, 2005Dec 20, 2011Ambrx, Inc.Process of producing non-naturally encoded amino acid containing high conjugated to a water soluble polymerUS8093356Jan 10, 2012Ambrx, Inc.Pegylated human interferon polypeptidesUS8097702Jan 17, 2012Ambrx, Inc.Modified human interferon polypeptides with at least one non-naturally encoded amino acid and their usesUS8114630Apr 30, 2008Feb 14, 2012Ambrx, Inc.Modified interferon beta polypeptides and their usesUS8119603Jan 28, 2005Feb 21, 2012Ambrx, Inc.Modified human interferon polypeptides and their usesUS8143216Oct 25, 2007Mar 27, 2012Ambrx, Inc.Modified human growth hormoneUS8163695Oct 25, 2007Apr 24, 2012AmbrxFormulations of human growth hormone comprising a non-naturally encoded amino acidUS8178108May 15, 2012Ambrx, Inc.Methods for expression and purification of recombinant human growth hormoneUS8178494Oct 25, 2007May 15, 2012Ambrx, Inc.Modified human growth hormone formulations with an increased serum half-lifeUS8232371Dec 8, 2011Jul 31, 2012Ambrx, Inc.Modified human interferon polypeptides and their usesUS8278418Sep 25, 2009Oct 2, 2012Ambrx, Inc.Modified animal erythropoietin polypeptides and their usesUS8383365Feb 26, 2013Ambrx, Inc.Methods of making FGF-21 mutants comprising non-naturally encoded phenylalanine derivativesUS8420792Sep 7, 2007Apr 16, 2013Ambrx, Inc.Suppressor tRNA transcription in vertebrate cellsUS8569233Jul 24, 2012Oct 29, 2013Eli Lilly And CompanyModified animal erythropoietin polypeptides and their usesUS8618257Sep 7, 2007Dec 31, 2013Ambrx, Inc.Modified human plasma polypeptide or Fc scaffolds and their usesUS8735539Aug 17, 2011May 27, 2014Ambrx, Inc.Relaxin polypeptides comprising non-naturally encoded amino acidsUS8778880Jan 28, 2005Jul 15, 2014Ambrx, Inc.Human growth hormone modified at position 35US8906676Jan 28, 2005Dec 9, 2014Ambrx, Inc.Modified human four helical bundle polypeptides and their usesUS8907064Oct 18, 2007Dec 9, 2014Ambrx, Inc.Modified human four helical bundle polypeptides and their usesUS8946148Nov 20, 2008Feb 3, 2015Ambrx, Inc.Modified insulin polypeptides and their usesUS9079971Jan 2, 2013Jul 14, 2015Ambrx, Inc.Modified FGF-21 polypeptides comprising non-naturally occurring amino acidsUS9121024Sep 28, 2009Sep 1, 2015Ambrx, Inc.Non-natural amino acid replication-dependent microorganisms and vaccinesUS9121025Jul 7, 2013Sep 1, 2015Ambrx, Inc.Non-natural amino acid replication-dependent microorganisms and vaccinesUS9133495Sep 7, 2007Sep 15, 2015Ambrx, Inc.Hybrid suppressor tRNA for vertebrate cellsUS9156899Sep 20, 2013Oct 13, 2015Eli Lilly And CompanyModified animal erythropoietin polypeptides and their usesUS9175083Oct 16, 2007Nov 3, 2015Ambrx, Inc.Antigen-binding polypeptides and their usesUS9260472Oct 18, 2007Feb 16, 2016Ambrx, Inc.Modified human four helical bundle polypeptides and their usesUS9434778Oct 23, 2015Sep 6, 2016Bristol-Myers Squibb CompanyModified FGF-21 polypeptides comprising an internal deletion and uses thereofUS20030177537 *Dec 20, 2002Sep 18, 2003Sembiosys Genetics Inc.Expression of epidermal growth factor in plant seedsUS20030208792 *Jan 7, 2003Nov 6, 2003The Scripps Research InstituteMethod for using tobacco mosaic virus to overproduce peptides and proteinsUS20060135427 *Dec 21, 2005Jun 22, 2006Ambrx, Inc.Formulations of human growth hormone comprising a non-naturally encoded amino acidUS20070065912 *Jul 21, 2006Mar 22, 2007Abbott LaboratoriesMultiple Gene Expression including sORF Constructs and Methods with Polyproteins, Pro-Proteins, and ProteolysisUS20080108792 *Oct 26, 2007May 8, 2008Ambrx, Inc.Human Interferon Molecules and Their UsesUS20080113411 *Oct 29, 2007May 15, 2008Ambrx, Inc.Modified Human Plasma Polypeptide or Fc Scaffolds and Their UsesUS20080113412 *Oct 29, 2007May 15, 2008Ambrx, Inc.Modified Human Plasma Polypeptide or Fc Scaffolds and Their UsesUS20080114154 *Oct 19, 2007May 15, 2008Ambrx, Inc.Modified Human Interferon Polypeptides and Their UsesUS20080119640 *Oct 26, 2007May 22, 2008Ambrx, Inc.Human Interferon Molecules and Their UsesUS20080132681 *Oct 26, 2007Jun 5, 2008Ambrx, Inc.Human Interferon Molecules and Their UsesUS20080153745 *Dec 1, 2005Jun 26, 2008Ambrx, In.Compositions of tRNA and uses thereofUS20080255045 *Mar 19, 2008Oct 16, 2008Ambrx, Inc.Modified FGF-21 Polypeptides and Their UsesUS20080317670 *Dec 13, 2006Dec 25, 2008Ambrx, Inc.Compositions Containing, Methods Involving, and Uses of Non-Natural Amino Acids and PolypeptidesUS20090137736 *Nov 8, 2006May 28, 2009Ambrx, Inc.Accelerants for the Modification of Non-Natural Amino Acids and Non-Natural Amino Acid PolypeptidesUS20090208454 *Apr 30, 2008Aug 20, 2009Ambrx, Inc.Modified interferon beta polypeptides and their usesUS20090222936 *Nov 6, 2006Sep 3, 2009Timothy RichmondRecombinant Expression of Multiprotein Complexes Using PolygenesUS20100035812 *Jul 22, 2009Feb 11, 2010Ambrx, Inc.Modified Bovine G-CSF Polypeptides And Their UsesUS20100093608 *Sep 25, 2009Apr 15, 2010Ambrx, Inc.Modified animal erythropoietin polypeptides and their usesUS20100135959 *Jun 2, 2006Jun 3, 2010Ambrx, Inc.Human Interferon Molecules and Their UsesUS20100159585 *Sep 7, 2007Jun 24, 2010Ambrx, Inc.Suppressor tRNA Transcription in Vertebrate CellsUS20100159586 *Sep 7, 2007Jun 24, 2010Ambrx, Inc.Hybrid Suppressor tRNA for Vertebrate CellsUS20100160212 *Sep 7, 2007Jun 24, 2010Ambrx, Inc.Modified Human Plasma Polypeptide or Fc Scaffolds and Their UsesUS20100167347 *Mar 15, 2010Jul 1, 2010Ambrx, Inc.Compositions of Aminoacyl-tRNA Synthetase and Uses ThereofUS20100173379 *Mar 15, 2010Jul 8, 2010Ambrx, Inc.Compositions of Aminoacyl-tRNA Synthetase and Uses ThereofUS20100173380 *Jul 8, 2010Ambrx, Inc.Compositions of Aminoacyl-tRNA Synthetase and Uses ThereofUS20100184140 *Mar 15, 2010Jul 22, 2010Ambrx, Inc.Compositions of Aminoacyl-tRNA Synthetase and Uses ThereofUS20100298212 *Nov 20, 2008Nov 25, 2010Ambrx, Inc.Modified Insulin Polypeptides and Their UsesUS20110144307 *Jun 16, 2011Ambrx, Inc.Methods and Compositions Comprising Non-Natural Amino AcidsUS20110150861 *Jun 23, 2011Abbott LaboratoriesSorf constructs and multiple gene expressionUS20110172401 *Jul 14, 2011Ambrx, Inc.Modified FGF-21 Polypeptides and Their UsesUS20110195483 *Sep 28, 2009Aug 11, 2011Ambrx, Inc.Non-Natural Amino Acid Replication-Dependent Microorganisms and VaccinesUS20110195899 *Aug 5, 2010Aug 11, 2011Ambrx, Inc.Formulations of Human Growth Hormone Comprising a Non-Naturally Encoded Amino AcidUS20110207914 *Aug 25, 2011Ambrx, Inc.Modified Human Plasma Polypeptide or Fc Scaffolds and Their UsesEP0552559A2 *Dec 21, 1992Jul 28, 1993Unilever PlcTransgenic plants resistant to microbian infectionEP0682709A1 *Jan 31, 1994Nov 22, 1995Board Of Regents The University Of Texas SystemFusion proteins including a cleavage site recognized by a plant virus proteaseEP0700999A2 *Jul 13, 1995Mar 13, 1996Sankyo Company LimitedExpression systems utilizing autolyzing fusion proteins and a novel reducing polypeptideEP2468768A2Jul 21, 2006Jun 27, 2012Abbott LaboratoriesMultiple gene expression including sorf contructs and methods with polyproteins, pro-proteins, and proteolysisEP2468881A2Jul 21, 2006Jun 27, 2012Abbott LaboratoriesMultiple gene expression including sorf contructs and methods with polyproteins, pro-proteins, and proteolysisEP2484774A2Jul 21, 2006Aug 8, 2012Abbott LaboratoriesMultiple gene expression including sorf contructs and methods with polyproteins, pro-proteins, and proteolysisWO1994010319A1 *Nov 4, 1993May 11, 1994MAX-PLANCK-Gesellschaft zur F�rderung der Wissenschaften e.V.Process for increasing the protein content of plantsWO1994016087A1 *Jan 12, 1994Jul 21, 1994Institut National De La Recherche AgronomiquePlant virus-resistant transgenic plants and method for producing sameWO1994018331A2 *Jan 31, 1994Aug 18, 1994Board Of Regents, The University Of Texas SystemFusion proteins including a cleavage site recognized by a plant virus proteaseWO1994018331A3 *Jan 31, 1994Oct 13, 1994Univ TexasFusion proteins including a cleavage site recognized by a plant virus proteaseWO1995012669A1 *Nov 1, 1994May 11, 1995The Texas A & M University SystemExpression of foreign genes using a replicating polyprotein producing virus vectorWO1995017514A1 *Dec 19, 1994Jun 29, 1995Zeneca LimitedExpression of self-processing polyproteins in transgenic plantsWO1995021249A1 *Feb 3, 1995Aug 10, 1995The Scripps Research InstituteA cassette to accumulate multiple proteins through synthesis of a self-processing polypeptideWO1995024486A1 *Mar 9, 1995Sep 14, 1995Zeneca LimitedANTIMICROBIAL PROTEINS FROM ARALIA AND $i(IMPATIENS)WO1996021019A1 *Jun 7, 1995Jul 11, 1996Asgrow Seed CompanyPapaya ringspot virus coat protein geneWO1996021029A1 *Dec 21, 1995Jul 11, 1996University Technologies International, Inc.Oil body proteins as carriers of high value proteinsWO1996021033A2 *Jun 7, 1995Jul 11, 1996Asgrow Seed CompanyPAPAYA RINGSPOT VIRUS NIa PROTEASE GENEWO1996021033A3 *Jun 7, 1995Oct 10, 1996Asgrow Seed CoPAPAYA RINGSPOT VIRUS NIa PROTEASE GENEWO2000011175A1 *Aug 17, 1999Mar 2, 2000Syngenta LimitedGenetic method for the expression of polyproteins in plantsWO2002061100A1 *Jan 30, 2001Aug 8, 2002Wisconsin Alumni Research FoundationExpression of multiple proteins in transgenic plants* Cited by examinerClassifications U.S. Classification800/301, 435/320.1, 800/298International ClassificationC12N15/62, C12N15/82, C12N9/50, C12N9/64, C07K14/08, C12N9/72Cooperative ClassificationC12Y304/23015, C12Y304/21069, C07K14/005, C12N2770/34022, C12N15/8241, C12N15/8216, C12N9/6459, C12N15/62, C12N9/506, C07K2319/00, C12N9/6486European ClassificationC12N9/64F23F, C12N9/64F21Q68, C07K14/005, C12N15/82B, C12N15/82C, C12N15/62, C12N9/50A1Legal EventsDateCodeEventDescriptionJan 5, 1995ASAssignmentOwner name: ASGROW SEED COMPANY CORP., MICHIGANFree format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:UPJOHN COMPANY, THE A DE CORP.;REEL/FRAME:007275/0211Effective date: 19941230Apr 23, 1996FPAYFee paymentYear of fee payment: 4May 6, 1997ASAssignmentOwner name: SEMINIS VEGETABLE 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