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Exogenous Peptide Replacement of Three Loops in Domain II of Cry1Ab Toxin Reduces Its Toxicity against Helicoverpa armigera (Lepidoptera: Noctuidae)

Lin, L., Shao, E-S., Shi, P., Huang, Z-P., Guan, X., Gelbic, I.
African entomology 2019 v.27 no.1 pp. 66-74
Escherichia coli, Helicoverpa armigera, binding capacity, brush border membrane vesicles, growth retardation, larvae, mutants, peptides, proteolysis, toxicity, toxins, trypsin
To investigate the contribution of surface-exposed loops (loops 1, 2 and 3) in the toxicity of Cry1Ab against Helicoverpa armigera larvae, three loop-replaced Cry1Ab mutants were prokaryotically expressed in Escherichia coli BL21 (DE3). Proteolytic processing of Cry1Ab toxins with trypsin indicated that the replacement of each surface-exposed loop caused no damage to toxin stability. Results of in vitro binding and competition assay indicated that, similarly to the native Cry1Ab toxin, the mutants retained almost the same binding affinity and specificity with brush border membrane vesicles (BBMV) of H. armigera. Nevertheless, H. armigera larvae fed with each loop-replaced Cry1Ab mutant showed less growth inhibition in comparison with those fed with native Cry1Ab toxin. The most significant decrease in toxicity was observed from larvae fed with loop 2-replaced Cry1Ab mutant, with a c. 12-fold increase in the weight of tested larvae relative to that of larvae fed native Cry1Ab toxin. The results demonstrate that the replacement of three surface-exposed loops located in domain II of Cry1Ab toxin may result in a certain reduction in the toxicity. Relative to loop 1 and loop 3, loop 2 seem to play a more important role in generating toxicity of Cry1Ab against H. armigera larvae.