Nucleic acid molecules which code for enzymes derived from wheat and which are involved in the synthesis of starch
View Patent ↗Nucleic acid molecules are described which encode enzymes involved in starch synthasis in plants. These enzymes are wheat isoamylases. The invention furthermore relates to vectors and host cells which contain the above-described nucleic acid molecules, in particular to transformed plant cells and plants which can be regenerated from these and which have an increased or reduced activity of the isoamylases according to the invention.
1. An isolated nucleic acid molecule encoding a protein with the function of a wheat isoamylase, selected from the group consisting of
(a) a nucleic acid molecule encoding a protein comprising the amino acid sequence of SEQ ID NO:3,
(b) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:2or a ribonucleotide sequence corresponding thereto; and
(c) a nucleic acid molecule whose nucleotide sequence deviates from the sequence of a nucleic acid molecule mentioned under (a) or (b) owing to the degeneracy of the genetic code,
wherein the isolated nucleic acid molecule is isolated from wheat.
2. An isolated nucleic acid molecule which hybridizes with the complement of the nucleic acid molecule of claim 1 , wherein the hybridization conditions comprise a hybridization temperature of 68° C., a hybridization buffer salt concentration of 5×SSC, a wash temperature of 68° C., and a wash buffer salt concentration of 0.5×SSC, wherein the isolated nucleic acid molecule encodes a wheat isoamylase.
3. The nucleic acid molecule as claimed in claim 1 which is a DNA molecule.
4. The nucleic acid molecule as claimed in claim 3 which is a cDNA molecule.
5. The nucleic acid molecule as claimed in claim 1 comprising regulatory elements.
6. The nucleic acid molecule as claimed in claim 1 which is an RNA molecule.
7. A vector containing the DNA molecule as claimed in claim 1 .
8. The vector as claimed in claim 7 , wherein said nucleic acid molecule is operably linked in sense orientation to regulatory elements which ensure transcription and synthesis of a translatable RNA in prokaryotic or eukaryotic cells.
9. A host cell which is transformed with the nucleic acid molecule as claimed in claim 1 , or transformed with a vector comprising the nucleic acid molecule of claim 1 , or a cell which is derived from the host cell, which derived cell comprises said nucleic acid molecule.
10. A process for the preparation of a protein encoded by the nucleic acid molecule as claimed in claim 1 , wherein a host cell transformed with the nucleic acid molecule of claim 1 or transformed with a vector comprising said nucleic acid molecule is cultured under conditions which permit said protein to be synthesized and said protein is isolated from the cultured cells and/or the culture medium.
11. A process for generating a transgenic plant cell, wherein
a) the nucleic acid molecule as claimed in claim 1 or
b) a vector comprising said nucleic acid molecule is integrated into the genome of a plant cell.
12. A transgenic plant cell which has been transformed with the nucleic acid molecule as claimed in claim 1 or transformed with a vector comprising the nucleic acid molecule of claim 1 , or a cell which is derived from the transgenic plant cell, which derived cell comprises said nucleic acid molecule.
13. A process for generating a transgenic plant cell, wherein
a1) the nucleic acid molecule as claimed in claim 1 or
a2) a vector comprising the nucleic acid molecule of claim 1 is integrated into the genome of a plant cell and
b) an intact plant is regenerated from said plant cell.
14. A plant containing the plant cell as claimed in claim 12 .
15. The plant as claimed in claim 14 which is a crop plant.
16. The plant as claimed in claim 15 which is a starch-storing plant.
17. The plant as claimed in claim 16 which is a monocotyledonous plant or maize.
18. The plant as claimed in claim 17 which is a barley, rye or wheat plant.
19. Propagation material of the plant as claimed in claim 14 , wherein said propagation material comprises said nucleic acid molecule.
20. A process for the production of strach comprising isolating starch frrom the cell as claimed in claim 12 , the plant as claimed in claim 14 or the propagation material as claimed in claim 19 .