Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se – Higher plant – seedling – plant seed – or plant part
Patent
1996-05-08
1999-03-09
Robinson, Douglas W.
Multicellular living organisms and unmodified parts thereof and
Plant, seedling, plant seed, or plant part, per se
Higher plant, seedling, plant seed, or plant part
4353201, 536 236, 8003202, A01H 500, C12N 1582, C12N 1563, C07H 2104
Patent
active
058803343
DESCRIPTION:
BRIEF SUMMARY
This application is a 371 of PCT/JP95/01356 filed Jul. 6, 1995.
TECHNICAL FIELD
The present invention relates to a phosphoenolpyruvate carboxykinase (hereinafter also referred to as "PCK") gene and to a recombinant vector containing the same.
BACKGROUND ART
PCK is an enzyme which reversibly catalyzes the reaction forming oxaloacetic acid by carboxylation of phosphoenolpyruvate. ATP-dependent PCK (EC 4.1.1.49) and GTP-dependent PCK (EC 4.1.1.32) are known. Plant PCKs are dependent on ATP and play an important role in the process of photosynthesis by which starch is formed from carbon dioxide.
On the other hand, C.sub.4 plants include mainly the plants belonging to family Gramineae originated in tropical zone, and are well adapted for strong sun light, high temperature and to shortage of water. More particularly, the rate of photosynthesis of C.sub.4 plants is twice that of C.sub.3 plants, and the photosynthesis is not inhibited by oxygen in the air. The photosynthesis of C.sub.4 plants does not reach saturation even if they are irradiated with a light having an intensity by which the photosynthesis of C.sub.3 plants is saturated. Further, the optimum temperature for photosynthesis of C.sub.4 plants is higher than that of C.sub.3 plants.
As mentioned above, PCK of plants plays an important role in photosynthesis. Thus, if a C.sub.3 plant is transformed so that it produces the PCK of C.sub.4 plant, it is expected that various effects may be obtained, such as increase in the rate of photosynthesis, efficient utilization of the sun light, and promotion of photosynthesis at a high temperature. However, so far, PCK gene of plants, needless to say C.sub.4 plants, has not been entirely sequenced.
DISCLOSURE OF THE INVENTION
Accordingly, an object of the present invention is to provide a cloned PCK gene of a C.sub.4 plant, a recombinant vector containing the gene, and a plant transformed with the recombinant vector.
The present inventors succeeded in cloning the PCK gene of Urochloa panicoides which is a C.sub.4 plant and in determining the entire sequence of the gene as well as deduced amino acid sequence encoded by the gene. The present inventors further succeeded in constructing a recombinant vector containing the PCK gene and in transforming a plant with the recombinant vector to obtain a transformed plant which expresses the PCK gene, thereby completing the present invention.
That is, the present invention provides a cloned DNA encoding the amino acid sequence shown in SEQ ID NOS: 1-6 in Sequence Listing or the same amino acid sequence as shown in SEQ ID NOS: 1-6 except that one or more amino acid is added, deleted, inserted or substituted, with the proviso that the polypeptide having this amino acid sequence has phosphoenolpyruvate carboxykinase activity. The present invention also provides a recombinant vector comprising the DNA according to the present invention, which can express said DNA in a host cell. The present invention further provides a plant transformed with the recombinant vector according to the present invention, which produces phosphoenolpyruvate carboxykinase.
By the present invention, the PCK gene of Urochloa panicoides was cloned and its nucleotide sequence was determined. It is expected that by introducing this gene into a C.sub.3 plant, the efficiency of photosynthesis may be promoted, photo energy may be more efficiently utilized and resistance to high temperature of the plant may also be promoted.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 shows the positions and lengths of the cDNAs employed for obtaining PCK1 which is a full length cDNA sequence of Urochloa panicoides;
FIG. 2 shows the positions and lengths of the cDNAs employed for obtaining PCK2 which is a full length cDNA sequence of Urochloa panicoides;
FIG. 3 shows the amino acid sequence encoded by PCK1 in comparison with that encoded by PCK2;
FIG. 4 is a gene map showing inserted DNA region of the recombinant vector used for transformation of rice plants; and
FIG. 5 is a schematic view showing localization of PCK prote
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Baba et
Arai Masao
Burnell James Nigel
Finnegan Patrick M.
Murai Nobuhiko
Suzuki Shoichi
Japan Tobacco Inc.
Robinson Douglas W.
Wai Thanda
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