Magnesium chelatase

Details Magnesium chelatase Magnesium chelatase

2000-06-01

2003-05-27

Bui, Phuong T. (Department: 1638)

Multicellular living organisms and unmodified parts thereof and

Method of introducing a polynucleotide molecule into or...

C435S006120, C435S069100, C435S183000, C435S410000, C435S419000, C435S252300, C435S320100, C530S350000, C530S370000, C536S023100, C536S023200, C536S023600, C536S024100, C536S024300, C800S295000

Reexamination Certificate

active

06570063

ABSTRACT:

FIELD OF THE INVENTION
This invention is in the field of plant molecular biology. More specifically, this invention pertains to nucleic acid fragments encoding magnesium chelatase subunit in plants and seeds.
BACKGROUND OF THE INVENTION
Magnesium chelatase catalyzes the insertion of the magnesium cation (Mg
2+
) into protoporphyrin IX, the branchpoint in the tetrapyrrole biosynthetic pathways leading to (bacterio)chlorophyll synthesis. In photosynthetic bacteria, magnesium chelatase activity requires three different subunits encoded by the genes bchD, bchH and bchI (Willows, R. D. and Beale, S. I., (1998)
J. Biol. Chem
. 273:34206-34213). It has been proposed that the BchH subunit initially forms a complex with protoporphyrin IX while the Bch I and BchD subunits form a complex in an ATP-dependent activation step. The BchI-BchD complex then inserts the magnesium cation into the BchH-bound protoporphyrin IX in an ATP-dependent reaction (Willows, R. D. and Beale, S. I. supra).
Similarly in higher plants, three distinct proteins, CHLD, CHLH, and CHLI, encoded by the genes ChlD, ChlH and ChlI respectively (Papenbrock J. et al., (1997)
Plant J
. 12:981-990) are required for magnesium chelatase activity. They share significant sequence similarity with their bacterial counterparts, further suggesting that the mechanism of magnesium chelation proceeds more or less similarly in plants and bacteria (Guo, R. et al., (1998)
Plant Physiol
. 116:605-615).
Since magnesium chelatase is an enzyme specific for chlorophyll synthesis, it presents a potential target for discovery and development of herbicides nontoxic to man and animals. Isolation of more genes encoding magnesium chelatase subunits provides a wider array of possible targets, thereby increasing the chances of successfully identifying promising herbicide candidates.
SUMMARY OF THE INVENTION
The present invention concerns an isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of: (a) a first nucleotide sequence encoding a polypeptide of at least 50 amino acids having at least 80% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs:14, 38, 42, and 48; (b) a second nucleotide sequence encoding a polypeptide of at least 100 amino acids having at least 80% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs:6, 10, and 30; (c) a third nucleotide sequence encoding a polypeptide of at least 100 amino acids having at least 85% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:26; (d) a fourth nucleotide sequence encoding a polypeptide of at least 100 amino acids having at least 90% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs:2, 20, and 34; (e) a fifth nucleotide sequence encoding a polypeptide of at least 100 amino acids having at least 95% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:24; (f) a sixth nucleotide sequence encoding a polypeptide of at least 130 amino acids having at least 85% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:28; (g) a seventh nucleotide sequence encoding a polypeptide of at least 150 amino acids having at least 80% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:32; (h) an eighth nucleotide sequence encoding a polypeptide of at least 250 amino acids having at least 85% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:44; (i) a ninth nucleotide sequence encoding a polypeptide of at least 250 amino acids having at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:22; 0) a tenth nucleotide sequence encoding a polypeptide of at least 380 amino acids having at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:8; (k) an eleventh nucleotide sequence encoding a polypeptide of at least 400 amino acids having at least 85% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:50; (1) a twelfth nucleotide sequence encoding a polypeptide of at least 400 amino acids having at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:4; (m) a thirteenth nucleotide sequence encoding a polypeptide of at least 750 amino acids having at least 90% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:12; (n) a fourteenth nucleotide sequence encoding a polypeptide of at least 1110 amino acids having at least 95% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:36; and (o) a fifteenth nucleotide sequence comprising the complement of (a), (b), (c), (d), (e), (f), (g), (h), (i), (0), (k), (1), (m), or (n).
In a second embodiment, it is preferred that the isolated polynucleotide of the claimed invention comprises a nucleotide sequence which comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 41, 43, 47, and 49 that codes for the polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 42,44, 48, and 50.
In a third embodiment, this invention concerns an isolated polynucleotide comprising a nucleotide sequence of at least one of 60 (preferably at least one of 40, most preferably at least one of 30) contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 41, 43, 47, and 49 and the complement of such nucleotide sequences.
In a fourth embodiment, this invention relates to a chimeric gene comprising an isolated polynucleotide of the present invention operably linked to at least one suitable regulatory sequence.
In a fifth embodiment, the present invention concerns a host cell comprising a chimeric gene of the present invention or an isolated polynucleotide of the present invention. The host cell may be eukaryotic, such as a yeast or a plant cell, or prokaryotic, such as a bacterial cell. The present invention also relates to a virus, preferably a baculovirus, comprising an isolated polynucleotide of the present invention or a chimeric gene of the present invention.
In a sixth embodiment, the invention also relates to a process for producing a host cell comprising a chimeric gene of the present invention or an isolated polynucleotide of the present invention, the process comprising either transforming or transfecting a compatible host cell with a chimeric gene or isolated polynucleotide of the present invention.
In a seventh embodiment, the invention concerns a magnesium chelatase subunit polypeptide selected from the group consisting of: (a) a polypeptide of at least 50 amino acids having at least 80% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs:14, 38, 42, and 48; (b) a polypeptide of at least 100 amino acids having at least 80% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs:6, 10, and 30; (c) a polypeptide of at least 100 amino acids having at least 85% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:26; (d) a polypeptide of at least 100 amino acids having at least 90% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs:2, 20, and 34; (e) a polypeptide of at least 100 amino acids having at least 95% identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:24; (f) a

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