Cloning of acryIIIA endotoxin gene ofBacillus thuringiensis var.tenebrionis and its transient expression inindica rice |
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Authors: | Tony M Johnson A S Rishi Pritilata Nayak Soumitra K Sen |
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Institution: | (1) Plant Molecular and Cellular Genetics and Centre for Plant Molecular Biology, Bose Institute, Centenary Building, P1/12, CIT Scheme VII-M, 700 054 Calcutta, India;(2) Present address: Department of Biochemistry, College of Biological Sciences, 140 Gortner Laboratory, 1479 Gortner Avenue, St. Paul, 55108-1022 MN, USA;(3) Biotechnology Division, Rallis India Limited, Rallis Agrochemical Research Station, 560 058 Bangalore, India |
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Abstract: | Damage caused to rice production by coleopteran insects like rice weevil (Sitophilus oryzae), a stored grain insect pest and rice hispa (Dicladispa armigera), a pest of the growing plant is quite high. In order to combat the damage, generation of insect resistant transgenic rice
plant was considered desirable. CryIIIA endotoxin ofBacillus thuringiensis var.tenebrionis, a 65 kDa protein toxic to coleopteran insects, figured as the candidate gene product. Thus, the cryIIIA gene was isolated
from a local isolate ofBacillus thuringiensis var.tenebrionis. The gene was tailored at the N-terminal end to its minimal size by using a synthetic ATG codon which replaced the first
codon next to ATG of threonine to proline. This modification did not affect the functional property of the gene product. A
chimeric construct of the modifiedcryIIIA gene was developed containing CaMV35S promoter andnos terminator for plant expression. The expressibility of thecryIIIA gene inindica rice was judged through test for transient expression in indica rice protoplasts. |
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Keywords: | Bt cryIIIA gene coleopteran rice pest N-terminal truncation sandwich ELISA transient gene expression |
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