Abstract
Bacillus thuringiensis, a well-known Gram-positive soil bacterium produces insecticidal proteins that accumulate in inclusion bodies during sporulation such as Cry and Cyt proteins. These inclusions are solubilized in insect midgut, releasing δ-endotoxins that exhibit insecticidal proteolytic activity. But there are several important insects such as lepidopteran black cutworm (BCW) that are less sensitive to their action. B. thuringiensis also produces proteins during their vegetative growth stage which are designated as VIP (Vegetative Insecticidal Protein) and are very effective against BCW. These second generation proteins are divided into four families Vip1, Vip2, Vip3, and Vip4 according to their amino acid sequence homology. Vip proteins from culture supernatant of Bacillus isolates can be purified by different methods and are screened for their insecticidal activity. The purification methods (1) anion exchange chromatography followed by SDS-PAGE gel electrophoresis and (2) Fast Protein Liquid Chromatography (FPLC) which are described here.
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References
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Senthilkumar, M., Amaresan, N., Sankaranarayanan, A. (2021). Purification and Characterization of Vips from Culture Supernatant of Bacillus. In: Plant-Microbe Interactions. Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1080-0_70
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DOI: https://doi.org/10.1007/978-1-0716-1080-0_70
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-1080-0
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