Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
  • C12N15/09—Recombinant DNA-technology
  • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
  • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
  • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
  • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
  • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
  • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
  • C12N15/8247—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified lipid metabolism, e.g. seed oil composition
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
  • C12N9/10—Transferases (2.)
  • C12N9/1025—Acyltransferases (2.3)
  • C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)

Definitions

• Figure 1 provides the 204 amino acid conserved sequence profile identified from comparisons of glycerol-3-phosphate acyltransferase and various lysophosphatidic acid acyltransferase using PSI-BLAST.
• polynucleotides of the invention can be used as a hybridization probe for RNA, cDNA, or genomic DNA to isolate full length cDNAs or genomic clones encoding a polypeptide and to isolate cDNA or genomic clones of other genes that have a high sequence similarity to a polynucleotide set forth in the Sequence Listing.
• Such probes will generally comprise at least 15 bases.
• Preferably such probes will have at least 30 bases and can have at least 50 bases.
• Particularly preferred probes will have between 30 bases and 50 bases, inclusive.
• Polypeptides of the present invention include isolated polypeptides encoded by a polynucleotide comprising a sequence selected from the group of a sequence contained in SEQ ID NOs: 1, 3, 5, 7, 9, 10, 12, 14, 16, 18, 20, 22, and 226-233.
• the inactive precursors generally are activated when the prosequences are removed. Some or all of the prosequences may be removed prior to activation. Such precursor protein are generally called proproteins.
• the polynucleotide and polypeptide sequences can also be used to identify additional sequences which are homologous to the sequences of the present invention. The most preferable and convenient method is to store the sequence in a computer readable medium, for example, floppy disk, CD ROM, hard disk drives, external disk drives and DVD, and then to use the stored sequence to search a sequence database with well known searching tools. Examples of public databases include the DNA Database of Japan (DDBJ)(http://www.ddbj.nig. ac.jp/); Genebank
• host cell is meant a cell which contains a vector and supports the replication, and/or transcription or transcription and translation (expression) of the expression construct.
• Host cells for use in the present invention can be prokaryotic cells, such as E. coli, or eukaryotic cells such as yeast, plant, insect, amphibian, or mammalian cells.
• host cells are monocotyledenous or dicotyledenous plant cells.
• antibodies to the acyltransferase protein can be prepared by injecting rabbits or mice with the purified protein or portion thereof, such methods of preparing antibodies being well known to those in the art. Either monoclonal or polyclonal antibodies can be produced, although typically polyclonal antibodies are more useful for gene isolation.
• Western analysis may be conducted to determine that a related protein is present in a crude extract of the desired plant species, as determined by cross- reaction with the antibodies to the acyltransferase protein. When cross-reactivity is observed, genes encoding the related proteins are isolated by screening expression libraries representing the desired plant species. Expression libraries can be constructed in a variety of commercially available vectors, including lambda gtl 1, as described in Sambrook, et al. (Molecular
• nucleic acid or amino acid sequences encoding an acyltransferase of this invention may be combined with other non-native, or “heterologous”, sequences in a variety of ways.
• heterologous sequences is meant any sequence which is not naturally found joined to the acyltransferase, including, for example, combinations of nucleic acid sequences from the same plant which are not naturally found joined together.
• the Ti- or Ri-plasmid containing the T-DNA for recombination may be armed (capable of causing gall formation) or disarmed (incapable of causing gall formation), the latter being permissible, so long as the vir genes are present in the transformed Agrobacterium host.
• the armed plasmid can give a mixture of normal plant cells and gall.
• ATAT9F GGATCCGCGGCCGCACAATGGGAGCTCAGGAGAAACGGCG 177
• ATAT2 pCGB8565
• ATAT3 pCGN8566
• P CGN8918 ATAT6
• pCGN8913 ATAT7
• pCGN8904 pCGN9970
• pCGN9940 ATAT10
• P CGN8567 ATATl 1
• pCGN8632 ATLPAATl
• P CGN9901 YSCAT1 also referred to as g>2132299).
• ⁇ CGN9902 (YSCAT2, also referred to as gi 1078509), pCGN9903 (YSCAT3, also referred to as gi2132939), pCGN9904 (YSCAT4, also referred to gi2133031), pCGN9905 (YSCAT5, also referred to as gi320748).
• pCGN9906 pCGN9906
• Fragments which were cloned into the pCGN8624 vector created the constructs pCGN8903 (ATATl), pCGN8573 (ATAT2), pCGN8911 (ATAT3), pCGN8598 (ATAT4), pCGN8921 (ATAT6), pCGN8916 (ATAT7), pCGN8907 (ATAT8), pCGN9971 (ATAT9), pCGN9944 (ATAT10), pCGN8577 (ATATl 1), and pCGN8635 (ATLPAATl) for the sense expression of the respective coding sequences from the 35S promoter.
• pCGN8903 pCGN8573
• pCGN8911 pCGN8911
• pCGN8598 ATAT4
• pCGN8921 ATAT6
• pCGN8916 pCGN8916
• pCGN8907 pCGN8907
• pCGN9944
• Transgenic Brassica plants are obtained by Agrobacterium-mediated transformation as described by Radke et ⁇ l (Theor. Appl Genet. (1988) 75:685-694; Plant Cell Reports (1992) 77:499-505).
• Transgenic Arabidopsis thaliana plants may be obtained by Agrobacterium-mediated transformation as described by Valverkens et al, (Proc. Nat. Acad. Sci. (1988) 55:5536-5540), or as described by Bent et al. ((X99A), Science 265: 1856-1860), or Bechtold et al. ((1993), C.R.Acad.Sci, Life Sciences 316: 1194-1199) or Clough, et al.

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• Micro-Organisms Or Cultivation Processes Thereof (AREA)
• Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
• Enzymes And Modification Thereof (AREA)

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• 1999
  • 1999-09-24 WO PCT/US1999/022231 patent/WO2000018889A2/en not_active Ceased
  • 1999-09-24 JP JP2000572337A patent/JP2002525105A/ja not_active Withdrawn
  • 1999-09-24 CA CA002343969A patent/CA2343969A1/en not_active Abandoned
  • 1999-09-24 EP EP99958636A patent/EP1115844A2/de not_active Withdrawn

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