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Cloning of an insecticidal toxin gene of Bacillus thuringiensis subsp. tenebrionis and its expression in Escherichia coli cells

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Jahn,  Norbert
Department of Molecular Biology, Max Planck Institute for Medical Research, Max Planck Society;

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Geider,  Klaus
Department of Molecular Biology, Max Planck Institute for Medical Research, Max Planck Society;

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Citation

Jahn, N., Schnetter, W., & Geider, K. (1987). Cloning of an insecticidal toxin gene of Bacillus thuringiensis subsp. tenebrionis and its expression in Escherichia coli cells. FEMS Microbiology Letters, 48(3), 311-315. doi:10.1111/j.1574-6968.1987.tb02615.x.


Cite as: http://hdl.handle.net/21.11116/0000-0008-091F-D
Abstract
Abstract A structural gene of a crystal protein toxic for coleoptera larvae was cloned from plasmid DNA of Bacillus thuringiensis subsp. tenebrionis (BTT). The DNA was partially digested with restriction enzyme BamHI and fragments were inserted into cosmid pHC79. In Western blot analysis extracts from infected Escherichia coli cells revealed expression of the BTT crystal protein in antibiotic‐resistant cells. Cell lysates from a selected E. coli clone were toxic for larvae of the Colorado potato beetle (Leptinotarsa decemlineata). The electrophoretic mobility in SDS gels of crystal protein from E. coli cells was 68 kDa and 74 kDa as observed for BTT‐toxin in B. thuringiensis extracts. The cosmids obtained were unstable during cellular propagation. The deletion product still carried the δ‐endotoxin gene.