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Transient Expression of Foreign Genes in Insect Cells sf9 for Protein Functional Assay
Published on: February 22, 2018
Vip3Aa induces apoptosis in cultured Spodoptera frugiperda (Sf9) cells
Kun Jiang1, Si-Qi Mei1, Ting-Ting Wang1
1Department of Microbiology, College of Life Sciences, Nankai University, Tianjin 300071, China.
Abstract:
The vegetative insecticidal proteins (Vip) secreted by many Bacillus thuringiensis strains during their vegetative growth stage are regarded as second generation insecticidal proteins, as they share no sequence or structural homology with known crystal insecticidal proteins (Cry) and have a broad insecticidal spectrum. Compared with insecticidal crystal proteins (ICPs), the insecticidal mechanisms of Vips have been little studied. Here we investigated the mechanism responsible for Vip3Aa toxicity in cultured insect cells. Using, flow cytometry analyzes, TUNEL staining and DNA fragmentation assays, we show that Vip3Aa can induce apoptosis in Spodoptera frugiperda (Sf9) cells and cause cells to arrest at the G2/M phase. We also show that Vip3Aa can disrupt mitochondrial membrane potential (ΔΨm), leading to the activation of Sf-caspase-1, suggesting that a mitochondrial mediated and caspase dependent pathway may be involved in Vip3Aa-induced apoptosis in Sf9 cells.

