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Biotechnol Lett. 2013 Nov;35(11):1889-94. doi: 10.1007/s10529-013-1275-6. Epub 2013 Jun 26.

Identification of a second cytotoxic protein produced by Bacillus thuringiensis A1470.

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Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center, 1465-5 Aikawa-machi, Kurume, Fukuoka, 839-0861, Japan,


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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