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Cloning and expression inEscherichia coli of entomotoxic protein gene fromBacillus thuringiensis subspecieskurstaki
Authors:Rakesh Tuli  Jyoti Saluja  Nihal K Notani
Institution:1. Molecular Biology and Agriculture Division, Bhabha Atomic Research Centre, 400 085, Trombay, Bombay, India
Abstract:A plasmid borne larvicidal crystal protein gene from B.thuringiensis subspecieskurstaki was cloned inEscherichia coli using a specific 20-mer oligonucleotide probe. The gene expressed inE. coli at a high level. TransgenicE. coli cells produced large irregular bodies which looked bright under phase contrast microscopy. The phase bright bodies released by sonic disruption of cells could be pelleted by centrifugation. Toxicity trials on the larvae ofSpodoptera litura showed that the pellet was antifeedant and toxic to the larvae. The supernatant was only mildly antifeedant. Even short term feeding of larvae on the toxin delayed the onset of pupation.
Keywords:Bacillus thuringiensis            delta-endotoxin  gene cloning and expression  lepidopteran toxin            Spodoptera toxin  parasporal crystal protein  toxin gene
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