Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide encodes an inhibitory rna molecule
Patent
1997-10-21
1999-10-26
Smith, Lynette R. F.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide encodes an inhibitory rna molecule
536 236, 536 245, 536 231, 435 691, 435468, 435410, 435418, 435419, 4353201, 800278, 800295, 800281, 8003202, C12N 1582, C12N 1584, C12N 504, C12N 1529
Patent
active
059732261
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a method for changing the composition of phospholipids produced by an organism and to a recombinant vector therefor.
BACKGROUND ART
Phospholipids are naturally occurring nonionic surface active agents which are valuable in the field of foods, medicines and production of various materials. Phospholipids are mainly produced from biological materials, and the composition of the phospholipids is one of the elements which determine the quality of the material.
The factor which participates in the determination of the composition of phospholipids is unknown and no methods by which the composition of phospholipids is changed by genetic engineering process have been reported.
DISCLOSURE OF THE INVENTION
An object of the present invention is to provide a method for artificially changing the composition of phospholipids produced by cells.
The present inventors intensively studied to discover that the composition of phospholipids produced by cells can be changed by inhibiting expression of the gene encoding phospholipase D (hereinafter also referred to as "PLD") which is a kind of phospholipase, thereby completing the present invention.
That is, the present invention provides a method for changing the composition of phospholipids produced by a host cell, comprising transforming said host cell with a recombinant DNA having an antisense gene of a phospholipase D gene, which antisense gene is expressed in said host cell to produce an mRNA that hybridizes with mRNA of a phospholipase D gene in said host cell thereby inhibiting expression of said phospholipase D gene. The present invention also provides a recombinant DNA having an antisense gene of a phospholipase D gene, which antisense gene is expressed in a host cell to produce an mRNA that hybridizes with mRNA of phospholipase D gene in said host cell thereby inhibiting expression of said phospholipase D gene.
BEST MODE FOR CARRYING OUT THE INVENTION
PLD is a kind of phospholipase, which, for example, catalyzes the reaction of hydrolyzing lecithin to liberate phosphatidic acid and choline. This enzyme is known to occur in plants, animals and microorganisms. The host cell employed in the method of the present invention may be any cell of plants, animals and microorganisms, as long as it has a PLD gene. The host cell is preferably a plant cell, more preferably a cell of a spermatophyte.
The term "spermatophyte" herein means plants which flower and form seeds. In the present invention, among the cells of spermatophytes, cells of monocotyledons, especially rice, are preferred.
The recombinant vector used for the method of the present invention comprises an antisense gene of the PLD gene, which antisense gene can inhibit expression of the PLD gene in the host cells. The nucleotide sequences of PLD genes are known. For example, the nucleotide sequences of PLD genes of microorganisms and animals are described in Japanese Laid-open Patent Application (Kokai) No. 3-187382; Adrian L. M. Hodgson, Phllip Bird, and Ian T. Nisbet 1990, "Cloning, nucleotide sequence, and expression in Escherichia coli of the Phospholipase D gene from Corynebacterium pseudotuberculosis" Journal of Bacteriology 172, 1256-1261; and Japanese Laid-open Patent Application (Kokai) No. 5-76357. As for PLD genes of plants, the nucleotide sequences of the PLD genes of rice and maize are described in WO95/09234. Further, PLD genes originated from Arabidopsis and castor bean have also been reported (Plant Physiol.(1995)109:1497-1499, Plant Gene Register PGR95-096, J. Biol. Chem. (1994) 269:20312-20317).
In the above-mentioned reference (Plant Gene Register PGR95-096), it is described that PLD genes originated from rice, maize, Arabidopsis and castor bean have high homologies each other, so that it is suggested that PLD genes of spermatophytes are well conserved. Therefore, PLD genes of spermatophytes may easily be obtained by a conventional method using the above-mentioned sequences of the PLD genes of rice, maize and the like as probes.
Thus, since the
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Morioka Shinji
Ueki Jun
Japan Tobacco Inc.
Smith Lynette R. F.
Zaghmout Ousama
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