Using phage display technology to obtain Crybodies active against non-target insects

Abstract

The insecticidal Cry toxins produced by Bacillus thuringiensis (Bt) are increasingly important in the biological control of insect pests and vectors of human disease. Markets for Bt products and transgenic plants expressing their toxins are driven by their specificity, safety and the move away from chemical control agents. However, the high specificity of Cry toxins can also prove to be a limitation when there is no known Cry toxin active against a particular target. Novel activities can be discovered by screening natural Bt isolates or through modifications of the Cry proteins. Here we demonstrate the use of λ-phage displaying Cry1Aa13 toxin variants modified in domain II loop 2 (Crybodies) to select retargeted toxins. Through biopanning using gut tissue from larvae of the non-target insect Aedes aegypti, we isolated a number of phage for further testing. Two of the overexpressed Cry toxin variants showed significant activity against A. aegypti larvae while another induced mortality at the pupal stage. We present the first report of the use of phage display to identify novel activities toward insects from distant taxonomic Orders and establish this technology based on the use of Crybodies as a powerful tool for developing tailor-made insecticides against new target insects.

Making 3D-Cry Toxin Mutants: Much More Than a Tool of Understanding Toxins Mechanism of Action

“…The latest report of a 3D-Cry toxin in vitro evolution using phage display technology was by Dominguez-Flores et al [140]. In this case, a library of "Crybodies" was displayed on a λ phage system similar to the one reported by Vilchez et al [130].…”

Um Olhar Principiológico

“…Em análise interdisciplinar, verificamos que a ciência econômica também observa o lucro como marca essencial da atividade empresarial, assumindo que "o objetivo crucial da empresa é a maximização do lucro 19 ". Parcela da doutrina privatista, inclusive, considera o lucro inserido no conteúdo material da função social da empresa, que seria a "obtenção do máximo de lucros possível, desde que atendidas às prescrições do ordenamento jurídico" 20 .…”

“…Fonte: KUCZYNSKI, E. Suicídio na Infância e Adolescência. Psicologia USP, v. 25, n. 3, p. 246-252, 2014. Estima-se que 800.000 pessoas morrem anualmente por suicídio, o que representa 1,4 % de mortes em todo o mundo, sendo registrado no ano de 2015 a prevalência no sexo feminino 0,00/100.000 habitantes (1-59 meses), 0,92/100.000 habitantes (5-14 anos de idade) e de 10,34/100.000 habitantes (15-29 anos de idade) e no sexo masculino 0,00/100.000 habitantes (1-59 meses), 1,10/100.000 habitantes (5-14 anos de idade) e de 14,06/100.000 (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29) O suicídio/autocídio é um fenômeno multifacetado de diferentes etiologias perpassando por inúmeros vieses de sua estrutura e de conhecimentos, como fatores que implicam o credo religioso, a política, a família, e fatores sociais além das disposições organopsíquicas que afetam a saúde mental, como a depressão (KUCZYNSKI, E.;, OLIVEIRA, B. 2016.…”

“…frugiperda, different studies have been conducted with this methodology, with particularly promising results against Callosobruchus maculatus, Acyrthosiphon pisum, Aphis gossypii, Myzus persicae, and Aedes aegypti [18][19][20]. Hence, PhD is an innovative agribio technological approach for pest control and was explored in this work.…”

Recombinant Phages for Spodoptera Frugiperda Control: New Perspectives to Tackle an Economic Problem

“…Oligopeptides can therefore be identified through an affinity selection strategy, making this tool extremely suitable for the identification of new molecules that act as biopesticides [17]. Although there are no reports of recombinant phage peptides for S. frugiperda, different studies have been conducted with this methodology, with particularly promising results against Callosobruchus maculatus, Acyrthosiphon pisum, Aphis gossypii, Myzus persicae, and Aedes aegypti [18][19][20]. Hence, PhD is an innovative agribiotechnological approach for pest control and was explored in this work.…”