Pengembangan Uji Toksisitas Kristal Protein Bacillus thuringiensis dengan Brush Border Membrane Vesicle

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Date
1998
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Balai Penelitian Bioteknologi Tanaman Pangan
Abstract
Use of Brush Border Membrane Visicle (BBMV) for Identification of Bt Crystal Protein Toxicity. T.P. Priyatno. Control of insect pest that live inside plant tissues is difficult. Development of transgenic plants carrying Bacillus thuringiensis (BQ genes for production of crystal protein is an alternative for effective control of these pests. The Bf genes that encode for production of toxic crystal protein is called the cry genes. Prior to transfering the genes to the target plant, they need to be identified and isolated from virulent Bf strains. For this purpose it needs an effective, efficient, and accurate technique. Toxicity test of Sf crystal protein by insect feeding is not effective for insect that live in plant tissues. A technique using brush border membrane vesicles (BBMV) isolated from epithelial cell membrane of an insect midgut could be used for this purpose. An active-Sf toxin is supposed to bind to a specific receptor located in the luminal plasmal membrane of the insect's midgut columnar epithelial cells. There is a positive correlation between binding affinity or number of binding sites and toxicity to the insect. BBMV that contain specific receptor need to bind the toxin. The BBMV technique could be done in vitro, therefore it supposed to be effective, efficient, and accurate. This technique can also be used for detection of insect resistance to Sf crystal protein. Development of an effective and efficient diagnostic kit for selection of virulent Bf strains and detection of insect resistance to Sf crystal protein is therefore necessary.
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Keywords
Brush border membrane vesicles (BBMV), insect pests, Bacillus thuringiensis strains, Bt crystal protein toxicity.
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