Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters fat – fatty oil – ester-type wax – or...
Patent
1996-11-18
1999-09-21
Robinson, Douglas W.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide alters fat, fatty oil, ester-type wax, or...
4351721, 4351723, 4352351, 435236, 530412, 536 2372, 800DIG9, 800DIG13, 800DIG25, 800DIG40, 800DIG42, 800DIG43, 800DIG44, A01H 500, C07K 114, C12N 701, C12N 704, C12N 1540, C12N 1582
Patent
active
059556471
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to methods for production of recombinant peptides and proteins. More particularly, this invention relates to techniques for inserting peptides into the coat protein of a virus, particularly for purposes of creating a vaccine.
2. Description of Related Art
Tobacco mosaic virus (TMV) is a well-characterized plant virus with a single, positive-sense RNA genome of 6395 nucleotides. The sequence of the TMV coat protein was initially derived back in the 1950's, and it was not until 1972 that the structure and roles of the forms of the TMV CP, its assembly and microscopic examination of the polymers were published (Durham, et al., J. Mol. Bio., 67:315-332 and 67:307-314). In 1982 the full genomic sequence of TMV RNA was published, and confirmation of the amino acid sequence of the CP as derived from the sequence of the viral RNA was confirmed (Goelet, et al., Proc. Natl. Acad. Sci. USA, 79:5818-5822, 1982). The 126 and 183-kDa proteins, which are required for virus replication (Ishikawa, et al. Nucleic Acids Res. 14:8291-8305, 1985) are translated directly from the genomic RNA from the same initiation codon. The 30 kDA movement protein (MP), which is involved in cell-to-cell movement, and the 17.5 kDA coat protein (CP) are translated from separate 3' coterminal subgenomic mRNAs (Siegel, et al., Proc. Nat. Acad. Sci. USA, 48:1845-1851, 1976; R. N. Beachy, et al., Virology, 63:84-97, 1975.
The life cycle of the TMV virus is well known. The amount of mRNA for the viral proteins determines the amount of each protein produced. The protein produced in the largest amount is the CP, which is as much as 5-10% of the total protein made in the infected cell. The structure of the TMV CP is reviewed in Butler, et al,. (J. of General Virology, 65:253-279, 1984). The CP encapsidates the viral RNA and is required for long-distance movement in the plant. The CP drives the assembly and encapsidation, which in turn enables systemic spread (Dawson, et al., Phytopathology, 78:783-789, 1988). Viral assembly and encapsidation are not required for local cell to cell spread of the viral RNA; however, the MP protein, a non-structural protein encoded by the virus, is required for local cell to cell spread of the viral RNA.
A number of research projects have been conducted to establish the use of TMV as a potential vector by inserting peptides into the coat protein. Even before the cloned cDNA of TMV was available, Haynes, et al. (Bio/Technology, 4:637-641, 1986) attached additional nucleotides at the 3' end of the coat protein mRNA, resulting in production of a fusion protein carrying sequences to serve as a vaccine for polio virus. The fusion CP was able to assemble and form rod-like structures in E. coli. Because the cDNA was unavailable, no work was done in a plant system.
The development of in vitro expression systems that allow production of infectious TMV RNAs from cloned full length cDNA genomes (Dawson, et al., Proc. Natl. Acad. Sci. USA, 83:1832-1836, 1986; Meshi, et al., Proc. Natl. Acad. Sci. USA, 83:5043-5047, 1986) has permitted the direct manipulation of the TMV genome at the DNA level. Highly infectious RNA transcripts of a full-length infectious cDNA clone of the U1 (common) strain of TMV have been produced in vitro using bacteriophage T7 RNA polymerase (Holt and Beachy, Virology 181:109-117, 1991) Thus, TMV RNA is a good candidate as a vector for the expression of foreign genes in plants. However, the TMV vectors developed using a CP gene modified to insert foreign genes usually fail to systemically express foreign genes, either through failure to produce intact CP for virus particle formation, or through loss of the foreign gene sequence during replication due to RNA recombination (N. Takamatsu, et al., EMBO J., 6:306-311, 1987; N. Takamatsu, et al, FEBS Lett., 2:73-76, 1989; W. O. Dawson, et al., Virology, 172:285-292, 1989). One exception is the TMV-ORSV (odontoglossum ringspot virus) hybrid vector, which includes an ORSV CP as an additional i
REFERENCES:
patent: 5618699 (1997-04-01), Hamamoto et al.
Hamamoto H, et al. "A new tobacco mosaic virus vector and its use for the systemic production of angiotensin-l-converting enzyme inhibitor in transgenic tobacco and tomato." Bio/Technology 11: 930-932, Aug. 1993.
Holt CA, et al. "In vivo complementation of infectious transcripts from mutant tobacco mosaic virus cDNAs in transgenic plants. " Virology 181: 109-117, 1991.
Citovsky, et al., Nuclear Localization of Agrobacterium VirE2 Protein in Plant Cells, Jun. 26, 1992, Science, 256: 1802-1805.
Bruening, et al., In Vitro and In Vivo Translation of the Ribonucleic Acids of a Cowpea Strain of Tobacco Mosaic Virus, 1976, Virology, 71:498-517.
Takamatsu, et al., Production of Enkephalin in Tobacco Protoplasts Unsing Tobacco Mosaic Virus RNA Vector, Aug. 1990, Federation of European Biochemical Societies, No. 1, 269:73-76.
Dawson, et al., Modifications of the Tobacco Mosaic Virus Coat Protein Gene Affecting Replication, Movement, and Symptomatology, 1988, The American Phytopathological Society, No. 6, 78:783-789.
Culver, et al., Virus-Host Interactions: Induction of Chlorotic and Necrotic Responses in Plant by Tobamoviruses, 1991, Annu. Rev. Phytopathol, 29:193-217.
Haynes, et al., Development of a Genetically-Engineered, Candidate Polio Vaccine Employing the Self-Assembling Properties of the Tobacco Mosaic Virus Coat Protein, Jul. 1986, Bio/Technology, 4:637-641.
Holt & Beachy, In Vivo Complementation of Infectious Transcripts from Mutant Tobacco Mosaic Virus cDNAs in Transgenic Plants, 1991, Virology, 181:109-117.
Hamamoto, et al., A New Tobacco Mosaic Virus Vector and its Use for the Systemic Production of Angiotensin-I-Converting Enzyme Inhibitor in Transgenic Tobacco and Tomato, Aug. 1993, Bio/Technology, vol. 11, pp. 930-932.
Beachy Roger N.
Fitchen John H.
Fitting Thomas
Holmes Emily
Nelson Amy J.
Robinson Douglas W.
The Scripps Research Institute
LandOfFree
Method for using tobacco mosaic virus to overproduce peptides an does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Method for using tobacco mosaic virus to overproduce peptides an, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Method for using tobacco mosaic virus to overproduce peptides an will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-82070