Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters fat – fatty oil – ester-type wax – or...
Reexamination Certificate
1999-07-23
2002-12-17
McElwain, Elizabeth F. (Department: 1638)
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide alters fat, fatty oil, ester-type wax, or...
C800S298000, C435S419000, C435S468000, C536S023600
Reexamination Certificate
active
06495739
ABSTRACT:
TECHNICAL FIELD
The present invention is directed to nucleic acid and amino acid sequences and constructs, and methods related thereto.
BACKGROUND
Through the development of plant genetic engineering techniques, it is possible to produce a transgenic variety of plant species to provide plants which have novel and desirable characteristics. For example, it is now possible to genetically engineer plants for tolerance to environmental stresses, such as resistance to pathogens and tolerance to herbicides. Another important example for such plant genetic engineering techniques is the production of valuable products in plant tissues, such as improved fatty acid compositions.
There is a need for improved means to obtain or manipulate fatty acid compositions and content, from biosynthetic or natural plant sources. For example, novel oil products, improved sources of synthetic triacylglycerols (triglycerides), alternative sources of commercial oils, such as tropical oils (i.e., palm kernel and coconut oils), and plant oils found in trace amounts from natural sources are desired for a variety of industrial and food uses.
To this end, the triacylglycerol (TAG) biosynthesis system in mammalian tissues, yeast and plants has been studied. In the cytoplasmic membranes of plant seed tissues which accumulate storage triglycerides (“oil”), fatty acyl groups are added sequentially by specific acyltransferase enzymes to the sn-1, sn-2 and sn-3 positions of glycerol-3-phosphate (G3P) to form TAG. This pathway is commonly referred to as the Kennedy or G3P pathway (FIG.
9
).
The first step in TAG formation is the acylation of the sn-1 position of glycerol-3-phosphate (G-3P), catalyzed by glycerophosphate acyltransferase (GPAT), to form lysophosphatidic acid (LA). The lysophosphatidic acid is subsequently acylated at the sn-2 position by lysophosphatidic acid acyltransferase (LPAAT) to create phosphatidic acid.
A key step in the formation of TAG is the dephosphorylation of the sn-3 position of phosphatidic acid (PA) to form sn-1,2-diacylglycerol (DAG) and inorganic phosphate catalyzed by the enzyme phosphatidic acid phosphatase (PAP, EC 3.1.3.4). The sn-1,2-diacylglycerol is acylated at the sn-3 position by diacylglycerol acyltransferase ultimately forming triacylglycerol (TAG).
The characterization of phosphatidic acid phosphatase (also known as PAP) from plants is useful for the further study of plant fatty acid synthesis systems and for the development of novel and/or alternative oils sources. Studies of plant mechanisms may provide means to further enhance, control, modify, or otherwise alter the total fatty acyl composition of triglycerides and oils. Furthermore, the elucidation of the factor(s) critical to the natural production of triglycerides in plants is desired, including the purification of such factors and the characterization of element(s) and/or cofactors which enhance the efficiency of the system. Of particular interest are the nucleic acid sequences of genes encoding proteins which may be useful for applications in genetic engineering.
SUMMARY OF THE INVENTION
The present invention provides nucleic acid sequences encoding for proteins which catalyze the dephosphorylation of phosphatidic acid (PA) to form sn-1,2-diacylglycerol (DAG). Such proteins are referred to herein as phosphatidic acid phosphatase (EC 3.1.3.4) or PAP.
By this invention, nucleic acid sequences encoding plant PAP may now be characterized with respect to enzyme activity. In particular, isolation of nucleic acid sequences encoding for PAP from Arabidopsis, Brassica, soybean and corn are provided.
Thus, this invention encompasses plant PAP nucleic acid sequences and the corresponding amino acid sequences, and the use of these nucleic acid sequences in the preparation of oligonucleotides containing PAP encoding sequences for analysis and recovery of plant PAP gene sequences. The plant PAP encoding sequence may encode a complete or partial sequence depending upon the intended use. All or a portion of the genomic sequence, or cDNA sequence, is intended.
Of special interest are recombinant DNA constructs which provide for transcription or transcription and translation (expression) of the plant PAP sequences. In particular, constructs which are capable of transcription or transcription and translation in plant host cells are preferred. For some applications a reduction in plant PAP may be desired. Thus, recombinant constructs may be designed having the plant PAP sequences in a reverse orientation for expression of an anti-sense sequence or use of co-suppression, also known as “transwitch”, constructs may be useful. Such constructs may contain a variety of regulatory regions including transcriptional initiation regions obtained from genes preferentially expressed in plant seed tissue. For some uses, it may be desired to use the transcriptional and translational initiation regions of the PAP gene either with the PAP encoding sequence or to direct the transcription and translation of a heterologous sequence.
In yet a different aspect, this invention relates to a method for producing a plant PAP in a host cell or progeny thereof via the expression of a construct in the cell. Cells containing a plant PAP as a result of the production of the plant PAP encoding sequence are also contemplated herein.
In addition, methods for increasing oil content in developing seed as well as methods for producing novel oil compositions in developing seeds of oil producing plants are contemplated.
Also considered in this invention are the modified plants, seeds and oils obtained by expression of the plant PAP sequences and proteins of this invention.
REFERENCES:
patent: WO 95/27791 (1995-10-01), None
Broun et al, “Catalytic Plasticity of Fatty Acid Modification Enzymes Underlying Chemical Diversity of Plant Lipids” Nov. 1998, Science vol. 282, pp. 1315-1317.*
Van De Loo et al, “An oleate 12-hydroxylase fromTicinus communisL. is a fatty acyl desaturase homog”, Jul. 1995, Proc. Natl. Acad. Science, vol. 92 pp. 6743-6747.*
De Luca, “Molecular characterization of secondary metabolic pathways”, 1993, AgBiotech News and Information vol. 5 No. 6 pp. 225n-229N.*
Bork, “Go Hunting in sequence databases but watch out for the traps”, 1996, TIG vol. 12 No. 10 pp. 425-427.*
Smith et al, The challenges of genome sequence annotation or “The devil is in the details”, 1997, Nature Biotechnology vol. 15 pp. 1222-1223.*
Doerks, “Protein annotation: detective work for function prediction”, 1998, TIG vol. 14 No. 6 pp. 248-250.*
Brenner, “Errors in genome annotation”, 1999, vol. 15 No. 4 pp. 132-133.*
Venter et al., “The Sequence of the Human Genome”Science,291:1304-1351 (Feb. 2001).
Woese et al., “Conservation of primary structure in 16S ribosomal RNA”Nature,254:83-85 (Mar. 1975).
Stymne, et al., “Triacylglycerol Biosynthesis”The Biochemistry of Plantsvol. 9 pp: 175-214 (1987).
Kai, et al., “Cloning and Characterization of Two human Isozymes of Mg2+-independent Phosphatidic Acid Phosphatase”,The Journal of Biological Chemistry(1997) vol. 272, No. 39 pp: 24572-24578.
Kai, et al., “Identification and cDNA Cloning of 335-kDa Phosphatidic Acid Phosphatase (Type 2) Bound to Plasma Membranes”The Journal of Biological Chemistry(1996) vol. 271, No. 31, pp: 18931-18938.
Kanoh, et al., “Phosphatidic acid phosphatase from mammalian tissues: discovery of channel-like proteins with unexpected functions”Biochimica et Biophysica Acta(1997) pp: 56-62.
Katagiri, et al., Abstract: “A Role of Phosphatidic Acid Phosphatase (PAP) in Cell Elongation in Arabidopsis thaliana” Annual Meeting of The Japanese Society of Plant Physiologists, May 3-5, 1998, p. 460 Abstract XP000863412.
Sun, et al., “Acyl Coenzyme A Preference of the Glycerol Phosphate Pathway in the Microsomes from the Maturing Seeds of Palm, Maize,and Rapeseed”Plant Physiol.(1988) pp: 56-60,.
Lohden, et al., “Triacylglycerol biosynthesis in developing seeds ofTropaeolum majusL. andLimnanthes douglasiiR. Br.” Planta (1992) 188: 215-224.
Berg, et al., “Purification of a phosphatase which hydrolyzes phosphatidic acid, a k
Lassner Michael W.
Ruezinsky Diane M.
Arnold & Porter
Calgene LLC
McElwain Elizabeth F.
Stierwalt Brian K.
LandOfFree
Plant phosphatidic acid phosphatases does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Plant phosphatidic acid phosphatases, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Plant phosphatidic acid phosphatases will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2956338