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
1996-04-10
1998-10-20
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...
4351723, 43525233, 4353201, 435419, 536 236, 800250, A01H 500, A01H 510, C12N 121, C12N 514, C12N 1529, C12N 1582
Patent
active
058248730
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
This application relates to novel DNA constructs, plant cells containing the constructs and plants derived therefrom having modified gene expression.
2. Description of Related Art
The modification of plant gene expression has been achieved by several methods. The molecular biologist can choose from a range of known methods to decrease or increase gene expression or to alter the spatial or temporal expression of a particular gene. For example, the expression of either specific antisense RNA or partial (truncated) sense RNA has been utilised to reduce the expression of various target genes in plants (as reviewed by Bird and Ray, 1991, Biotechnology and Genetic Engineering Reviews 9:207-227). These techniques involve the incorporation into the genome of the plant of a synthetic gene designed to express either antisense or sense RNA. They have been successfully used to down-regulate the expression of a range of individual genes involved in the development and ripening of tomato fruit (Gray et al, 1992, Plant Molecular Biology, 19:69-87). Methods to increase the expression of a target gene have also been developed. For example, additional genes designed to express RNA containing the complete coding region of the target gene may be incorporated into the genome of the plant to "over-express" the gene product. Various other methods to modify gene expression are known; for example, the use of alternative regulatory sequences.
SUMMARY OF THE INVENTION
In work leading to the present invention we have identified a gene which expresses an enzyme involved in the ripening of tomatoes. This gene is hereinafter referred to as TOM41.
According to the preset invention there is provided DNA comprising a TOM41 DNA sequence. A TOM41 DNA sequence is any sequence encoding a ripening-related enzyme which is homologous to the sequence shown as SEQ ID NO 1. The DNA sequence may be derived from cDNA, from genomic DNA or may be synthesized ab initio.
A cDNA clone encoding the TOM41 gene product has been obtained from a ripening tomato cDNA library using the method described by Slater et al (1985, Plant Molecular Biology, 5:137-147). The clone pTOM41 includes a sequence coding for substantially the whole of the mRNA produced by the corresponding TOM41 gene.
The mRNA for which pTOM41 codes is expressed in ripening tomato fruit. pTOM41 mRNA was also detected in green fruit but not in ripening fruit of the rin (ripening inhibitor) mutant (Picton PhD Thesis, University of Nottingham 1988). Sequence hybridising to pTOM41 have been located in a single region of chromosome 1 of tomato (Kinzer et al, 1990, Theor Appl Genet, 79:489-496). The biochemical function of the products of these genes has not hitherto been fully elucidated.
The nucleotide sequence of the TOM41 cDNA is given as SEQ ID NO 1. The sequence is 1423 bases in length with a reading frame from 78 to 1280. Searches in DNA data bases indicate that the pTOM41 clone shows homology (50% amino acid sequence homolgy) to an open-reading frame (ORF2) associated with the prs gene of E. coli (EMBL Database accession M77237). This open-reading frame forms part of a minor tri-cistronic transcript with the gene for phosphoribosylpyrophosphate synthetase (Post DA et al, 1991, FASEB Journal 5, A812). ORF2 encodes a 31 kD protein the function of which is unknown. Phosphoribosylpyrophosphate acts as a ribosyl donor for synthesis of nucleotides as well as histidine and tryptophan.
The clone pTOM41 was deposited at The National Collections of Industrial and Marine Bacteria (23 St Machar Drive, Aberdeen, Scotland, AB2 1RY) under the terms of the Budapest Treaty on 18 Mar., 1993 under the accession number NCIMB 40543.
An alternative source of the DNA sequence is a suitable gene encoding the TOM41 gene product (the TOM41 enzyme). This gene may differ from the corresponding cDNA in that introns may be present. The introns are not transcribed into mRNA (or, if so transcribed, are subsequently cut out). Oligonucleotide probes or the cDNA clone may
REFERENCES:
Gao J, et al. "The stability of foreign protein production in genetically modified plant cells." Plant Cell Rep. 10: 533-536, 1991.
Liaw G-J, et al. "Characterization of downstream elements in a Raf-1 pathway." PNAS 90: 858-862, Feb. 1993.
Hiatt A, et al. "Production of antibodies in transgenic plants." Nature 342: 76-78, Nov. 2, 1989.
Koziel MG, et al. "Optimizing expression of transgenes with an emphasis on post-transcriptional events." Plant Mol. Biol. 32: 393-405, 1996.
Stam M, et al. "The silence of genes in transgenic plants." Ann. Bot. 79: 3-12, 1997.
Smith CJS, et al. "Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes." Nature 334: 724-726, 1988.
Grierson Donald
Lawrence Susan Dale
Moore Gloria Andrews
Schuch Wolfgang Walter
Nelson Amy J.
Robinson Douglas W.
University of Florida
Zeneca Limited
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