Abstract
The cry10Aa gene from the Brazilian Bacillus thuringiensis subsp israelensis S1804 strain was introduced into the genome of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in order to evaluate its expression in insect cells and its toxicity to A. grandis. The recombinant virus (vSyncry10Aa) was amplified in Trichoplusia ni (BTI-Tn5B1-4) cells and used to infect Spodoptera frugiperda larvae. Total extracts from S. frugiperda infected with the recombinant viruses were analyzed by SDS-PAGE and showed the presence of a polypeptide around 85 kD. Cuboid-shaped protein crystals were observed in insect extracts by light and scanning electron microscopy. Bioassays, using the recombinant virus infected-insect extracts, showed high toxicity to A. grandis larvae, with a LC50 of 7.12 µg/mL. Furthermore, a competition binding assay with the recombinant biotin-labeled Cry10A protein and brush border membrane vesicles (BBMV) from A. grandis indicated that the toxin binds specifically to BBMVs. Therefore, the Cry10A protein has a potential to be used in transgenic cotton plants for the control of this important insect pest. Keywords Bacillus thuringiensis; Toxicity; Recombinant protein
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