Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se – Higher plant – seedling – plant seed – or plant part
Reexamination Certificate
1998-08-20
2002-12-24
Riley, Jezia (Department: 1637)
Multicellular living organisms and unmodified parts thereof and
Plant, seedling, plant seed, or plant part, per se
Higher plant, seedling, plant seed, or plant part
C800S312000, C800S260000, C800S320300, C435S006120, C435S091100, C435S091200, C435S440000, C435S468000
Reexamination Certificate
active
06498286
ABSTRACT:
BACKGROUND OF THE INVENTION
The present invention is directed to germplasm containing an identifier nucleotide sequence and to a method for identifying germplasm, more particularly the source of germplasm. The present invention is especially useful for determining the ownership of germplasm, including plants and animals, or the ownership of germplasm used as a parent in the development of germplasm. Briefly, the invention is directed to the production of transgenic organisms which contain an identifier nucleotide sequence within the organellar genome, including but not limited to plastids and mitochondria. The identifier nucleotide sequence is selected such that it is capable of indicating the source of the germplasm. The organellar genome of germplasm to be tested is isolated and analyzed for the presence of the identifier nucleotide sequence. The presence of the identifier nucleotide sequence establishes the source of the germplasm.
The publications and other materials used herein to illuminate the background of the invention, and in particular, cases to provide additional details respecting the practice, are incorporated by reference, and for convenience are referenced in the following text by author and date and are listed alphabetically by author in the appended bibliography.
The classification of organisms has traditionally been done along more or less arbitrary and some-what artificial lines. For example, the living world has been divided into two kingdoms, namely plants and animals. This classification works well for generally familiar organsims, but becomes more difficult for such organisms as unicellular ones. Where classification of organisms becomes more than a scientific exercise is in the identification of plants and animals for hybridization and breeding programs, and in accurate and reliable identification of microorganisms which may infect plants and animals. For example, the plant breeder may wish to have a quick and reliable means of identifying different species and strains for use in their breeding programs. In addition, the owner of a particular plant variety may wish to have a quick and reliable means of identifying his variety for establishing derivation or ownership of germplasm. Thus, the correct identification of species or varieties of organisms is of particular importance.
Biological samples have been collected and analyzed for a variety of reasons relating to agricultural forensic, e.g., to identify individuals for population genetics or variety derivation. The known methods of analyzing biological samples from plants include using the proteins, specific enzymes or the nucleic acids to identify individually specific patterns within a species. For example, protein from a plant can be isolated and subjected to a two-dimensional electrophoretic analysis to produce a protein fingerprint (Anderson et al., 1985; Choi et al., 1984). Alternatively, isozymes, i.e., specific enzymes which exist in several forms in plants, can be isolated and subjected to electophoresis to produce an isozyme fingerprint (Nielsen, 1985; Shields et al., 1983). Finally, the nucleic acids can be isolated and analyzed to produce a DNA fingerprint (U.S. Pat. No. 5,674,687; U.S. Pat. No. 4,963,663; Helentjaris et al., 1985; Landry et al., 1985; Jeffreys et al., 1985). DNA fingerprinting has utilized restriction fragment length polymorphisms and the use of minisatellite and microsatellite probes and/or amplification primers to obtain highly specific patterns which are useful to identify individuals within a known species or in pedigree studies to determine lineage. DNA fingerprinting is generally useful for identifying individuals because it uses patterns that are individually-specific and complex. Similarly, two-dimensional protein fingerprint could be used to identify individuals on the basis of complex patterns. An isozyme fingerprint is less complex but also has less resolution power.
Although each of these methods or a combination of these methods can be used to identify individuals, they have several disadvantages including: (a) time-consuming, (b) complex, sometimes with many steps, (c) requiring skilled and knowledgeable technicians, (d) non-standardization with results varying from lab to lab and test to test and (e) inaccurate measurements often with subjective interpretation. Thus, there is a definite need for a method to identify the source or parentage of germplasm which is easy to perform, relatively fast, does not require significant expertise and requires no subjectivity of interpretation. The present invention solves this need as illustrated herein.
SUMMARY OF THE INVENTION
The present invention is directed to germplasm containing an identifier nucleotide sequence and to a method for identifying germplasm, more particularly the source or parentage of germplasm. The present invention is especially useful for determining the ownership of germplasm, including plants and animals, or the ownership of germplasm used as a parent in the development of germplasm. Briefly, the invention is directed to the production of transgenic organisms which contain an identifier nucleotide sequence within the organellar genome. The identifier nucleotide sequence is selected such that it is capable of indicating the source of the germplasm. The organellar genome of germplasm to be tested is isolated and analyzed for the presence of the identifier nucleotide sequence. The presence of the identifier nucleotide sequence establishes the source or parentage of the tested germplasm.
DETAILED DESCRIPTION OF THE INVENTION
The present invention avoids the problems of prior art fingerprinting techniques for the identification of the source of germplasm. The present invention is simpler and easier to use than prior art fingerprinting techniques and requires no subjective interpretation of the results of the method.
The present invention is directed to germplasm containing an identifier nucleotide sequence and to a method for identifying germplasm, more particularly the source or parentage of germplasm. The present invention is especially useful for determining the ownership of germplasm, including plants and animals, or the ownership of germplasm used as a parent in the development of germplasm. Briefly, the invention is directed to the production of transgenic organisms which contain an identifier nucleotide sequence within the organellar genome. The identifier nucleotide sequence is selected such that it is capable of indicating the source of the germplasm. The organellar genome of germplasm to be tested is isolated and analyzed for the presence of the identifier nucleotide sequence. The presence of the identifier nucleotide sequence establishes the source or parentage of the tested germplasm.
In order to more fully understand the present invention, the following definitions are provided.
“Amplification of Polynucleotides” utilizes methods such as the polymerase chain reaction (PCR), ligation amplification (or ligase chain reaction, LCR) and amplification methods based on the use of Q-beta replicase. Also useful are strand displacement amplification (SDA), thermophilic SDA, nucleic acid sequence based amplification (3SR or NASBA) and repair chain reaction (RCR). These methods are well known and widely practiced in the art. See, e.g., U.S. Pat. Nos. 4,683,195 and 4,683,202 and Innis et al., 1990 (for PCR); Wu et al., 1989 and EP 320,308A (for LCR); U.S. Pat. Nos. 5,270,184 and 5,455,166 and Walker et al., 1992 (for SDA); Spargo et al., 1996 (for thermophilic SDA) and U.S. Pat. No. 5,409,818, Fahy et al., 1991 and Compton, 1991 for 3SR and NASBA. Reagents and hardware for conducting PCR are commercially available. Primers useful to amplify the identifier nucleotide sequence are preferably complementary to, and hybridize specifically to the identifier nucleotide sequence or to regions that flank a target region therein.
“Germplasm” refers to species or variety of a plant or an animal.
“Identifier nucleotide Sequence” refers to a unique nucleic acid which is not present either in the organellar genom
FFR Cooperative
Riley Jezia
Rothwell, Figg Ernst & Manbeck, PC
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