Nucleic acid molecules encoding wheat enzymes involved in starch synthesis
View Patent ↗The invention relates to nucleic acid molecules which code for enzymes and which are involved in the synthesis of starch in plants. These enzymes concern isoamylases derived from wheat. The invention also relates to vectors and host cells which contain the described nucleic acid molecules, especially transformed plant cells and plants which can be regenerated therefrom, which exhibit an increased or reduced activity of the inventive isoamylases.
1. A vector containing 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 shown under SEQ ID NO:3,
(b) a nucleic acid molecule comprising the nucleotide sequence shown under SEQ ID NO:2 or a ribonucleotide sequence corresponding hereto;
(c) a nucleic acid molecule which hybridizes under stringent conditions with a nucleic acid molecule mentioned under (a) or (b) or is complementary thereto, wherein hybridization 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; and
(d) a nucleic acid molecule whose nucleotide sequence deviates from the sequence of a nucleic acid molecule mentioned under (a), (b) or (c) owing to the degeneracy of the genetic code,
the nucleic acid molecule mentioned under (a), (c) and (d) having sequence identity of over 90% with SEQ ID NO:2, wherein said nucleic acid molecule is linked in sense orientation to regulatory elements, which ensure the synthesis of an untranslatable RNA in prokaryotic or eukaryotic cells.
2. A vector containing 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 shown under SEQ ID NO:3,
(b) a nucleic acid molecule comprising the nucleotide sequence shown under SEQ ID NO:2 or a ribonucleotide sequence corresponding hereto;
(c) a nucleic acid molecule which hybridizes under stringent conditions with a nucleic acid molecule mentioned under (a) or (b) or is complementary thereto, wherein hybridization 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; and
(d) a nucleic acid molecule whose nucleotide sequence deviates from the sequence of a nucleic acid molecule mentioned under (a), (b) or (c) owing to the degeneracy of the genetic code,
the nucleic acid molecule mentioned under (a), (c) and (d) having sequence identity of over 90% with SEQ ID NO:2, wherein said nucleic acid molecule is linked in antisense orientation to regulatory elements, which ensure the synthesis of an untranslatable RNA in prokaryotic or eukaryotic cells.
3. A host cell which is transformed with the vector of claim 1 or 2 or a cell which is derived from the host cell and comprises the vector of claim 1 or 2 .
4. A process for generating a transgenic plant tell, wherein the vector of claim 1 or 2 is integrated into the genome of a plant cell.
5. A transgenic plant cell that has been transformed with the vector of claim 1 or 2 or a cell which is derived from the transgenic plant cell and comprises the vector of claim 1 or 2 .
6. A plant containing the plant cell of claim 5 .
7. A process for generating a transgenic plant, wherein
(a) the vector of claim 1 or 2 is integrated into the genome of a plant cell and
(b) an intact plant is regenerated from said plant cell.
8. A transgenic plant generated by the process of claim 7 .
9. The plant of claim 6 , which is a crop plant.
10. The plant of claim 6 , which is a starch-synthesizing or starch-storing plant.
11. A propagation material of the plant of claim 6 .