Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se
Reexamination Certificate
2000-02-09
2004-11-02
Bui, Phuong T. (Department: 1638)
Multicellular living organisms and unmodified parts thereof and
Plant, seedling, plant seed, or plant part, per se
C435S006120, C435S069100, C435S183000, C435S410000, C435S419000, C435S252300, C435S320100, C530S370000, C536S023100, C536S023200, C536S023600, C536S024100, C536S024300, C536S024330, C536S024500, C800S278000
Reexamination Certificate
active
06812382
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 proteins involved in chloroplast division in plants and seeds.
BACKGROUND OF THE INVENTION
In plants plastid organelles harbor several metabolic pathways essential for plant growth, development and viability. Specifically, plastids contain the metabolic pathways for photosynthesis, the synthesis of various amino acids, lipids and plant growth regulatory hormones (Mullet, J. E. (1988)
Annu. Rev. Plant Physiol. Plant Mol Biol
. 39:475-502). During cell division plastids divide and segregate to daughter cells in a manner that insures that each daughter cell receives sufficient numbers of plastids to provide the cell with efficient photosynthetic and biosynthetic capabilities. Plastids appear to divide by a binary fission mechanism that involves two contractile rings, one on the stromal face of the inner envelope and one on the cytosolic face of the outer envelope (Kuroiwa, T., et al., (1998)
Int. Rev. Cyto
. 181:1-41). However, little is known about the exact nature of these rings or the precise mechanism of plastid division.
A genetic study of plastid division in Arabidopsis has revealed several arc mutations (accumulation and replication of chloroplasts) that define at least seven nuclear genes important in the control of plastid division (Pyke, K. A., (1997)
Am. J Bot
. 84:1017-1027). Osteryoung et al. have discovered, in the nuclear genome of Arabidopsis, a homolog of the bacterial cell division gene ftsZ. In bacteria defects in the ftsZ gene (for filament temperature-sensitive) cause filament formation and incomplete septum formation at the restrictive temperature (Osteryoung, K. W., et al., (1995)
Nature
376:473-474. The encoded FtsZ protein appears to be part of the cell division apparatus, specifically a cytoskelatal component of the contractile ring (Osteryoung, K. W., et al., (1998)
The Plant Cell
10:1991-2004) and Strepp et al. reports that mutations in the FtsZ gene resulted in the disruption of chloroplast division (Strepp, R., et al., (1998)
Proc. Natl. Acad. Sci
. 95:4368-4373).
Recent studies indicate that Arabidopsis contains two FtsZ genes one localized to the cytosol (FtsZ2) and one to the chloroplast (FtsZ1). Inhibition of either causes a severe decrease in the numbers of chloroplasts in Arabidopsis leaf cells (Osteryoung, K., et al., (1998)
The Plant Cell
10:1991-2004). Thus the two FtsZ genes are both essential for chloroplast division and appear to play distinct roles in the division process. The Osteryoung studies indicate further that plant FtsZ genes are grouped into two families and that the FtsZ proteins within a family share amino acid sequence identities ranging from 76 to 91% however between families the similarity declines to 61%.
There is a great deal of interest in identifying the genes that encode proteins involved in plastid division in plants. These gene may be used to enhance plastid transformation. For example, if gene expression of the ftsZ genes could be inhibited, plants with 1-3 very large chloroplasts per cell would be produced. By decreasing the number of chloroplasts per cell and increasing the size of each chloroplast a much better recipient cell for plastid transformation would result. Furthermore, because the FtsZ proteins are essential for chloroplast division, their inhibition could significantly decrease the, efficiency of photosynthesis and other metabolic pathways. Accordingly, the availability of nucleic acid sequences encoding all or a portion of a FtsZ protein would facilitate studies to better understand chloroplast division in plants and provide genetic tools to inhibit or otherwise alter chloroplast division in plants.
SUMMARY OF THE INVENTION
The present invention relates to isolated polynucleotides comprising a nucleotide sequence encoding a polypeptide of at least 145 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of corn cytosol and chloroplast associated FtsZ polypeptides of SEQ ID NOs:2 and 8, a rice cytosol FtsZ polypeptide of SEQ ID NO:10, a soybean chloroplast associated FtsZ polypeptide of SEQ ID NO:4, and wheat cytosol and chloroplast associated FtsZ polypeptides of SEQ ID NOs:6 and 14. The present invention also relates to an isolated polynucleotide comprising the complement of the nucleotide sequences described above.
The present invention also relates to isolated polynucleotides comprising a nucleotide sequence encoding a polypeptide of at least 241 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a soybean cytosol associated FtsZ polypeptide of SEQ ID NO:12. The present invention also relates to an isolated polynucleotide comprising the complement of the nucleotide sequences described above.
The present invention also relates to isolated polynucleotides comprising a nucleotide sequence encoding a polypeptide of at least 94 amino acids that has 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:16, 18, 20, 22 and 24. The present invention also relates to an isolated polynucleotide comprising the complement of the nucleotide sequences described above.
It is preferred that the isolated polynucleotides of the claimed invention consists of a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21 and 23 that codes for the polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24. The present invention also relates to an isolated polynucleotide comprising a nucleotide sequences 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, 15, 17, 19, 21, 23, and the complement of such nucleotide sequences.
The present invention relates to a chimeric gene comprising an isolated polynucleotide of the present invention operably linked to suitable regulatory sequences.
The present invention relates to an isolated 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.
The present invention relates to a process for producing an isolated 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 an isolated compatible host cell with a chimeric gene or isolated polynucleotide of the present invention.
The present invention relates to a cytosol and chloroplast associated FtsZ polypeptide of at least 145 amino acids comprising at least 90% homology based on the Clustal method of alignment compared to a polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10 and 14.
The present invention relates to a cytosol associated FtsZ polypeptides polypeptide of at least 241 amino acids comprising at least 90% homology based on the Clustal method of alignment compared to a polypeptide of SEQ ID NO:12.
The present invention relates to a method of selecting an isolated polynucleotide that affects the level of expression of a cytosol or chloroplast associated FtsZ polypeptide in a host cell, preferably a plant cell, the method comprising the steps of: (a) constructing an isolated polynucleotide of the present invention or an isolated chimeric gene of the present invention; (b) introducing the isolated polynucleotide or the isolated chimeric gene into a host cell; (c) measuring the level a cy
Hitz William D.
Miao Guo-Hua
Bui Phuong T.
E.I. du Pont de Nemours and Company
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