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
1995-03-20
1997-03-04
Fox, David T.
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...
800DIG44, 536 236, 536 241, 435 691, 435 701, 4351723, 435194, 435411, 435419, A01H 100, C12N 504, C12N 1529, C12N 1582
Patent
active
056081502
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
Recent advances in genetic engineering have provided the requisite tools to transform plants to contain foreign genes. It is now possible to produce plants which have unique characteristics of agronomic and crop processing importance. The ability to chose the tissues in which to express such foreign genes and the time during plant growth to obtain expression is possible through the choice of a regulatory sequence which turns on the gene, called the promoter. A wide range of promoters are known for various plants, plant tissues, and developmental stages.
The tomato is a very important plant for genetic engineering. It is readily transformed to express foreign genes and has many characteristics which are known to be improved by certain genes. It is also a valuable crop plant in many countries and was the first transgenic food crop approved for sale in the U.S.
Promoters useful in expressing foreign genes in tomato and other fruits are known. For example, the solids content of tomato fruit can be increased by expressing an ADPglucose pyrophosphorylase gene behind a fruit specific promoter. (Kishore, PCT Appl. WO 91/19806). The promoter from the 2A11 genomic clone (Pear, et al. (1989) Plant Mol. Biol. 13:639-651) will control expression of ADPglucose pyrophosphorylase in tomato fruit. The E4 and E8 promoters (Deikman, et al. (1988) The EMBO Journal 7:3315-3320), as well as the promoter for polygalacturonase are known to be fruit specific. However, the last three are limited to expression during a late stage in the development of the tomato fruit and so are known as red fruit promoters. The 2A11 promoter will cause expression during early stages, but is weaker than desired for some genes. Therefore, there is a need for stronger promoters which will cause expression of a gene during the development of the green fruit.
It is an object of the present invention to provide such promoters. It is a further object of the present invention to provide promoters which will function as fruit-specific in tomatoes and other fruit-bearing crops. It is a still further object of the present invention to provide DNA constructs containing these promoters and a gene encoding a desired protein or the antisense sequence for a less desirable protein.
SUMMARY OF THE INVENTION
The present invention provides two fruit-specific promoters which provide for expression at greater levels during early development of the fruit body of a plant. The two promoters are (1) TFM7 which is a DNA fragment of about 2.3 kb, of which 1.4 kb of the 3' end is shown in SEQ ID NO:1; and (2) TFM9 which is a DNA fragment of about 900 bp, of which bp of the 3' end is shown in SEQ ID NO:2.
The present invention also provides a recombinant, double-stranded DNA molecule comprising in sequence: which encodes a desired protein; and transcriptional termination and the addition of polyadenylated nucleotides to the 3' end of the RNA sequence, Plant cells and whole plants containing this DNA construct are also provided.
Plants in which this DNA construct may be used include, but are not limited to, tomato, strawberry, and raspberry.
DETAILED DESCRIPTION OF THE INVENTION
The expression of a plant gene which exists in double-stranded DNA form involves transcription of messenger RNA (mRNA) from one strand of the DNA by RNA polymerase enzyme, and the subsequent processing of the mRNA primary transcript inside the nucleus. This processing involves a 3' non-translated region which adds polyadenylate nucleotides to the 3' end of the RNA.
Transcription of DNA into mRNA is regulated by a region of DNA usually referred to as the promoter. The promoter region contains a sequence of bases that signals RNA polymerase to associate with the DNA, and to initiate the transcription of mRNA using one of the DNA strands as a template to make a corresponding complimentary strand of RNA.
Novel fruit specific promoters exhibiting high and specific expression during the development of the tomato fruit have been isolated. A differential screening approach util
REFERENCES:
Wingate et al. 1991. Plant Physiol. 97: 496-501.
Bonner Grace L.
Fox David T.
Hoerner, Jr. Dennis R.
Monsanto Company
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