Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide confers pathogen or pest resistance
Patent
1997-11-17
1999-06-22
Robinson, Douglas W.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide confers pathogen or pest resistance
435418, 435419, 800288, 800301, 800317, 8003173, A01H 500, C12N 510, C12N 1582
Patent
active
059144483
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to an antiviral plant producing lactoferrin and process for the preparation thereof. More specifically, it pertains to an antiviral plant producing lactoferrin, which is transformed with a lactoferrin (Lf) gene; a process for the preparation of the antiviral plant, which comprises transforming a plant with an expression vector containing a lactoferrin gene and culturing the transformed plant.
BACKGROUND OF THE INVENTION
Hitherto, various attempts have been made to develop a plant resistant to viral infection (hereinafter, an antiviral plant) by employing recombinant DNA technologies. For example, Powell Abel et al. introduced a cDNA encoding the coat protein of a virus to a plant (Infect Immun., 35, 792-796 (1986)), while the introduction of a defective viral replicase to a plant was attempted by Anderson et al. (P.N.A.S., 89, 8759-8763 (1992)). However, these methods are not practical in that, in order for a plant to have antiviral properties against a number of viruses, all the target viral genes must be introduced to the plant.
To solve this problem, Lodge et al. reported a method for conferring a general antiviral property on a plant by introducing thereto a gene encoding an antiviral protein, e.g., RIP (ribosome inhibiting protein) (P.N.A.S., 90, 7089-7093 (1993)). Although the plant transformed by this method shows an antiviral activity against various viruses, there exists the problem that the antiviral protein may be harmful to human, and hence, the application of the above method may be restricted.
Lactoferrin is a conjugated protein found in mammals, e.g., in human milk, saliva, tear, semen and leukocyte. It affects the growth and differentiation of cells and has an antibacterial and immunopotentiative activity (Arold et al., Science, 197, 263-265 (1977)). Further, it shows an antiviral activity against Friend virus (Lu et al., Cancer Res., 47, 4184-4188 (1987)), human immunodeficiency virus (HIV) and human cytomegalovirus (HCMV) (Meijer et al., J. Disease, 172, 380-388 (1995)).
Mitra and Zhang reported a method for providing a plant having an antibacterial activity by introducing thereto a cDNA encoding human lactoferrin (Plant Physiol., 106, 977-981 (1994)). They confirmed the production of human lactoferrin in a tobacco cell by conducting a western blotting analysis of a tobacco cell transformed with human lactoferrin cDNA and anti-human lactoferrin antibody. They also reported that the proliferation of bacteria was significantly suppressed when the extract of the transformed tobacco cell was added to a culture of bacteria, e.g., Pseudomonas sp., which is pathogenic to tobacco. However, Mitra and Zhang did not, or failed to produce a transformed tobacco plant from the transformed tobacco cells. Moreover, human lactoferrin produced in the transformed tobacco cell was defective, i.e., significant parts thereof were missing, while the antibacterial activity was confirmed only in vitro. Accordingly, the question of whether or not an antiviral plant can be produced from a transformed tobacco cell was left unanswered in the prior art.
SUMMARY OF THE INVENTION
Accordingly, it is an object of the present invention to provide an antiviral plant producing lactoferrin.
Another object of the present invention is to provide a process for preparing said antiviral plant.
A further object of the present invention is to provide a process for producing lactoferrin in a plant.
In accordance with one aspect of the present invention, there is provided a process for preparing an antiviral plant producing lactoferrin, which comprises transforming a plant with an expression vector containing lactoferrin (Lf) gene.
BRIEF DESCRIPTION OF THE DRAWINGS
The above and other objects and features of the present invention will become apparent from the following description of the invention, when taken in conjunction with the accompanying drawings, in which:
FIG. 1 shows the structure of plasmid pLSM1 comprising human lactoferrin (HLf) gene;
FIG. 2 depicts the resul
REFERENCES:
Mitra et al. Expression of a human lactoferrin cDNA in tobacco cells produces antibacterial protein(s). Plant Physiology. 106:977-981, Nov. 1994.
Lodge et al. Broad-spectrum virus resistance in transgenic plants expressing pokeweed antiviral protein. PNAS USA. 90(15):7089-7093, Aug. 1993.
"Proposed Mechanisms for the Involvement of Lactoferrin in the Hydrolysis of Nucleic Acids", TW Hutcherns et al., New York, 1994.
Lee Kyung-Kwang
Lee Mi-Hee
Liu Jang-Ryol
Yu Dae-Yeul
Korea Institute of Science and Technology
Robinson Douglas W.
Wai Thanda
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