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Interaction of Bombyx mori aminopeptidase N and cadherin-like protein with Bacillus thuringiensis Cry1Ac toxin / 生物工程学报
Chinese Journal of Biotechnology ; (12): 1809-1822, 2018.
Article in Chinese | WPRIM | ID: wpr-776288
ABSTRACT
Bacillus thuringiensis (Bt) produces Cry toxins that are widely used as insecticides in agriculture and forestry. Receptors are important to elucidate the mode of interaction with Cry toxins and toxicity in lepidopteran insects. Here, we purified the Cry toxin from Bt and identified this toxin by flight mass spectrometry as Cry1Ac, and then recombinantly expressed aminopeptidase N (BmAPN6) and repeat domains of cadherin-like protein (CaLP) of B. mori. Using co-immunoprecipitation (co-IP), Far-Western blotting, and enzyme-linked immunosorbent assays (ELISAs), we identified the interaction between Cry1Ac and BmAPN6. Furthermore, analysis of the cytotoxic activity of Cry1Ac toxin in Sf9 cells showed that BmAPN6 directly interacted with Cry1Ac toxin to induce morphological aberrations and cell lysis. We also used co-IP, Far-Western blotting and ELISAs to analyze the interactions of Cry1Ac with three binding sites corresponding to cadherin repeat (CR) 7 CR11, and CR12 of CaLP. Notably, the three repeat domains were essential Cry1Ac binding components in CaLP. These results indicated that BmAPN6 and CaLP served as a functional receptor involved in Bt Cry1Ac toxin pathogenicity. These findings represent an important advancement in our understanding of the mechanisms of Cry1Ac toxicity and provide promising candidate targets for gene editing to enhance resistance to pathogens and increase the economic value of B. mori.
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Full text: Available Index: WPRIM (Western Pacific) Main subject: Bombyx / Bacillus thuringiensis / Bacterial Proteins / Cadherins / CD13 Antigens / Endotoxins / Hemolysin Proteins / Larva / Metabolism Type of study: Prognostic study Limits: Animals Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2018 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Bombyx / Bacillus thuringiensis / Bacterial Proteins / Cadherins / CD13 Antigens / Endotoxins / Hemolysin Proteins / Larva / Metabolism Type of study: Prognostic study Limits: Animals Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2018 Type: Article