Abstract
Bacillus thuringiensis A1470 produces multiple proteins with similar molecular masses (~30 kDa) with cytotoxicity against human cell lines. One that was previously identified, parasporin-4, is a β-pore-forming toxin. The N-terminal sequence of a second cytotoxic protein was identical to a partial sequence of parasporin-2 produced by B. thuringiensis A1547. PCR was performed on total plasmid DNA from A1470 by using primers for parasporin-2 to amplify a gene which was then cloned. The cloned gene differed from A1547 parasporin-2 by 8 bp and the predicted protein differed by four amino acids. The gene was expressed in Escherichia coli, and the cytotoxic activities of the recombinant protein against four human cell lines (MOLT-4, Jurkat, HeLa, and HepG2) were similar to those of A1547 parasporin-2. We then confirmed that the A1470 strain simultaneously produces parasporin-2 and parasporin-4.
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Acknowledgments
We thank Associate professor Dr. Sakae Kitada at the Kyushu Institute of Technology and Professor Dr. Akio Ito at Kyushu University for anti parasporin-2 antibody and their helpful advice. We also thank Dr. Naoya Wasano for his invaluable advice on this study.
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Okumura, S., Ishikawa, T., Saitoh, H. et al. Identification of a second cytotoxic protein produced by Bacillus thuringiensis A1470. Biotechnol Lett 35, 1889–1894 (2013). https://doi.org/10.1007/s10529-013-1275-6
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DOI: https://doi.org/10.1007/s10529-013-1275-6