Variantes de inibidores de [alfa]-amilases selecionados por Phage Display com atividade para [alfa]-amilases do bicudo do algodoeiro (Anthonomus grandis, coleptera: curculionidae)




The cotton boll weevil Anthonomus grandis is an important insect-pest, that causes severe damage in cotton floral fruits and buds. The A. gradis larvae development control is a difficult job because it occurs inside the cotton floral structures. Starch granules from cotton structure are digested by a-amylases secreted to the intestinal lumen. However, a-amylase inhibitors (a-AIs) may be used in transgenic plants to control insect-pest. In this work, a large combinatorial library (107) of a-amylase inhibitors variants obtained by DNA shuffling of a-AI1 and a-AI2 homologous genes was used to select a-amylase inhibitor mutant genes against cotton boll weevil a- amylases (AGA). Twenty six a-AIs mutants were selected showing different mutation patterns. Five out of these mutants were expressed in Arabidopsis thaliana and their a- amylase inhibitory activity assayed. The a-AIs mutants were actives against AGA in a in vitro assay inhibiting at nM range. These results demonstrate the potential of a-AIs mutants in controlling of cotton boll weevil making them candidates to obtain pest resistant transgenic plants.


bicudo do algodoeiro anthonomus grandis combinatorial library a-amylase inhibitors anthonomus grandis inibidores de a-amilases biologia molecular biblioteca combinatória transgenic plants dna shuffling cotton boll weevil phage display arabidopsis thaliana

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