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
1999-09-21
2002-10-29
Fox, David T. (Department: 1638)
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
C800S288000, C800S287000, C800S292000, C800S293000, C800S294000, C800S317300, C435S320100, C435S468000, C435S469000, C435S470000, C435S418000, C435S419000, C435S414000, C435S421000, C536S023530, C536S023400
Reexamination Certificate
active
06472587
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a process for the production of herbicide-tolerant plants by expressing an exogenous herbicide-binding polypeptide in plants or plant organs. The invention furthermore relates to the use of the corresponding nucleic acids which encode a polypeptide, an antibody or parts of an antibody with herbicide-binding properties in transgenic plants, and the thus transformed plant itself.
2. Description of the Related Art
It is known that genetic engineering methods allow the specific transfer of foreign genes into the genome of a plant. This process is termed transformation, and the resulting plants transgenic plants. Transgenic plants are currently being employed in various fields of biotechnology. Examples of insect-resistant plants (Vaek et al. Plant Cell 5 (1987), 159-169), virus-resistant plants (Powell et al. Science 232 (1986), 738-743) and ozone-resistant plants (van Camp et al. BioTech. 12 (1994), 165-168). Examples of improved quality characteristics achieved by genetic engineering are: improved shelf life of fruit (Oeller et al. Science 254 (1991), 437-439), increased starch production in potato tubers (Stark et al. Science 242 (1992), 419), changes in starch (Visser et al. Mol. Gen. Genet. 225 (1991), 289-296) and lipid composition (Voelker et al. Science 257 (1992), 72-74), and production of foreign polymers (Poirer et al. Science 256 (1992), 520-523).
An important target of work carried out in the field of plant molecular genetics is the generation of herbicide tolerance. Herbicide tolerance is characterized by an improved compatibility (in terms of type or level) of the plant or plant organs with the herbicide applied. This can be effected in various ways. The known methods are utilization of a metabolic gene, for example the pat gene, in connection with glufosinate resistance (WO 8705629) or a target enzyme which is resistant to the herbicide, such as in the case of enolpyruvyl shikimate-3-phosphate synthase (WO 9204449), which is resistant to glyphosate, and the use of a herbicide in cell and tissue culture for the selection of tolerant plant cells and resulting resistant plants, such as described in the case of acetyl-CoA-carboxylase inhibitors (U.S. Pat. No. 5,162,602, U.S. Pat. No. 5,290,696).
Antibodies are proteins as component of the immune system. A joint feature of all antibodies is their spatial, globular structure, the construction of light and heavy chain and their basic capability of binding molecules or parts of a molecular structure with high specificity (Alberts et al., in: Molekularbiologie der Zelle [Molecular Biology of the Cell], 2nd Edition 1990, VCH Verlag, ISBN
3-527-27983-0, 1198-1237
). On the basis of these properties, antibodies have been utilized for a number of tasks. Application can be divided into application of the antibodies within the animal and human organisms in which they are produced, that is to say the so-called in-situ applications, and the ex-situ applications, ie. utilization of the antibodies after they have been isolated from the producing cells or organisms (Whitelam und Cockburn, TIPS Vol.1, 8 (1996), 268-272).
The use of somatic hybrid cell lines (hybridomas) as a source of antibodies against very specific antigens is based on work carried out by Köhler and Milstein (Nature 256 (1975) 495-97). This process allows so-called monoclonal antibodies to be produced which have a uniform structure and which are produced by means of cell fusion. Spleen cells of an immunized mouse are fused with mouse myeloma cells. This gives hybridoma cells which proliferate infinitely. At the same time, the cells secrete specific antibodies against the antigen with which the mouse had been immunized. The spleen cells provide the capability of antibody production while the myeloma cells contribute the capacity of unlimited growth and continuous secretion of antibodies. Since each hybridoma cell, being a clone, is derived from a single B cell, all antibody molecules produced have the same structure, including the antigen binding site. This method has greatly promoted the use of antibodies since antibodies which have a single, known specificity and a homogeneous structure are now available in unlimited quantities. Monoclonal antibodies are used widely in immunodiagnostics and as therapeutics.
In recent years, the so-called phage display method has become available for the production of antibodies, and here the immune system and the various immunizations in the animal are avoided. The affinity and specificity of the antibody are made to measure in vitro (Winter et al., Ann. Rev. Immunol. 12 (1994), 433-455; Hoogenboom TIBTech Vol 15 (1997), 62-70). Gene segments which contain the sequence which encodes the variable region of antibodies, ie. the antigen binding site, are fused with genes for the coat protein of a bacteriophage. Then, bacteria are infected with phages which contain such fusion genes. The resulting phage particles are now equipped with coats containing the antibody-like fusion protein, the antibody-binding domain pointing outward. Such a phage display library can now be used for isolating the phage which contains the desired antibody fragment and which binds specifically to a certain antigen. Each phage isolated in this manner produces a monoclonal antigen-binding polypeptide which corresponds to a monoclonal antibody. The genes for the antigen binding site, which are unique for each phage, can be isolated from the phage DNA and employed for constructing complete antibody genes.
In the field of crop protection, antibodies were utilized in particular as analytical tools ex-situ for the qualitative and quantitative detection of antigens. This includes the detection of plant constituents, herbicides or fungicides in drinking water (Sharp et al. (1991) ACS Symp Ser., 446 (Pestic. Residues Food Saf.) 87-95), soil samples (WO 9423018) or in plants or plant organs, and the utilization of antibodies as auxiliaries for the purification of bound molecules.
The production of immunoglobulins in plants was first described by Hiatt et al., Nature, 342 (1989), 76-78. The spectrum encompasses single-chain antibodies up to multimeric secretory antibodies (J. Ma and Mich Hein, 1996, Annuals New York Academy of Sciences, 72-81).
More recent attempts utilize antibodies in-situ for defending plants against pathogens, in particular viral diseases, by expressing, in plant cells, specific antibodies or parts thereof which are directed against viral coat proteins (Tavladoraki et al., Nature 366 (1993), 469-472; Voss et al., Mol. Breeding 1 (1995), 39-50).
An analogous approach has also been utilized for defending the plant against infection by nematodes (Rosso et al., Biochem Biophys Res Com, 220 (1996) 255-263). There exist examples for an application in pharmacology where the in-situ expression of antibodies in plants is utilized for oral immunization (Ma et al., Science 268 (1995), 716-719; Mason and Arntzen, Tibtech Vol 13 (1996), 388-392). The body is provided with antibodies formed by the plant and originating from plants or plant organs which are suitable for consumption, via the mouth, throat or digestive tract, which antibodies cause efficient immunoprotection. Moreover, a single-chain antibody against the low-molecular-weight plant hormone abscisic acid has already been expressed in plants, and a reduced availability of plant hormones, due to binding of abscisic acid in the plant, has been observed (Artsaenko et al., The Plant Journal 8 (5) (1995) 754-750).
Chemical control of weeds in agronomically important crops requires the use of highly selective herbicides. However, in some cases it is difficult to develop sufficiently selective herbicides which do not cause damage of the plant which provides the yield in any crop. The introduction of herbicide-resistant or -tolerant crop plants can contribute to solving this problem.
The development of herbicide-resistant crop plants by tissue culture or seed mutagenesis and natural selection is limited. Only those
Conrad Udo
Lerchl Jens
Möller Achim
Rabe Udo
Schiffer Helmut
BASF - Aktiengesellschaft
Fox David T.
Keil & Weinkauf
Kruse David H
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