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
Patent
1996-12-20
2000-05-30
Smith, Lynette R. F.
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
435243, 4352523, 4353201, 435419, 5303917, 536 2353, 800279, A01H 500, A01H 510, C07H 2104, C12N 121, C12N 514, C12N 1574, C12N 1582
Patent
active
060693010
DESCRIPTION:
BRIEF SUMMARY
The invention is drawn to antibodies that bind to insect gut proteins and their use, particularly their use in creating new hybrid toxin molecules. The invention further relates to microorganisms, plant cells and plants producing the said antibodies and hybrid toxins, respectively. The invention also encompasses insecticidal compositions and the use thereof in protecting plants against insect pests.
BACKGROUND OF THE INVENTION
Control of various pests through the use of biological molecules has been possible in only a limited number of cases. The best known examples of biological molecules with pesticidal uses are the .delta.-endotoxins from Bacillus thuringiensis (Bt). Various strains of Bt are known which produce insecticidal proteins, the .delta.-endotoxins, during sporulation. Some of these .delta.-endotoxins have useful insecticidal activities against different insect pests. However, use of the .delta.-endotoxins is limited because they are active against only a very few of the many insect pests.
The limited specificity of the Bt endotoxins is dependent, at least in part, on both the activation of the toxin in the insect gut (Haider, M. Z. et al., 1986, Eur. J. Biochem. 156:531-540) and its ability to bind to specific receptors present on the insect's midgut epithelial cells (Hofmann, C. P. et al., 1988, PNAS 85:7844-7848). Among the factors which prevent activity of a particular Bt .delta.-endotoxin against a specific insect is the lack of appropriate receptors in the insect gut or lack of affinity of the .delta.-endotoxin for the receptors which may be present, thus resulting in no binding of the .delta.-endotoxin to the brush border membranes. Therefore, the ability to control a specific insect pest using Bt .delta.-endotoxins at present depends on the ability to find an appropriate .delta.-endotoxin with the desired range of activity. In many cases, no such .delta.-endotoxin is known, and it is not certain that one even exists. For example, thousands of Bt strains have been screened for activity against western corn rootworm (WCRW), a major pest of maize. However, to date there are no reports of strains of Bt which produce a .delta.-endotoxin that is highly effective against WCRW.
Individual .delta.-endotoxins typically have a very narrow spectrum of activity, each being active against only one or a few insect pests. Moreover, the .delta.-endotoxins have been shown to be active against only a few members of but a small number of Orders of insects. The ability to produce additional proteins with unique pesticidal activities creates more options for the control of agricultural pests, particularly insects, using biological molecules with a high level of safety for non-target organisms. Thus, there is a need for binding proteins which can be designed to target a particular insect pest.
SUMMARY OF THE INVENTION
Hence, the present invention is drawn to antibodies, but especially to monoclonal antibodies or fragments thereof which bind to brush border membrane vesicles of the insect gut and to the gene or genes which encode these proteins. The antibodies according to the invention bind to proteins in the gut of a target insect especially to a target insect selected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, and Trichoptera but do not bind to mammalian brush border membranes or to plant microsomes. In particular the invention relates to a monoclonal antibody or fragment which binds to the gut of the western corn root worm. In a preferred embodiment, the invention relates to a monoclonal antibody which is selected from the group consisting of 2B5, 3B1, 10B6, 17F6, 14G1 and 16E4.
Also comprised by the invention are hybridoma cell lines producing the monoclonal antibodies according to the invention, but in particular hybridoma cell lines which have been deposited under accession number ATCC HB 11616, HB 11617, HB 11618, HB 11619 and HB 11620.
It is a further object of the inven
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Carozzi Nadine Barbara
Koziel Michael Gene
Meigs J. Timothy
Nelson Amy J.
Novartis Finance Corporation
Pace Gary M.
Smith Lynette R. F.
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