Toxicity and synergism in transgenic escherichia coli expressing four genes from Bacillus thuringiensis subsp. israelensis

The genes cyt1Aa and p20, encoding, respectively, cytolytic and accessory proteins of Bacillus thuringiensis subsp. israelensis, were introduced into previously constructed clones expressing cry4Aa and cry11Aa in Escherichia coil (Ben-Dov et al., 1995). Fifteen clones with all possible combinations of the four genes were obtained and found to express the genes included. Two new combinations, pVE4-ADRC and pVE4-ARC, expressing cyt1Aa, p20 and cry4Aa, with or without cry11Aa, respectively, were more toxic than their counterparts without cyt1Aa. They displayed the highest toxicity against Aedes aegypti larvae ever reached in transgenic bacteria. Five out of the six clones (except pVE4-DC) containing cry4Aa or cry11Aa (with or without p20) displayed varying levels of synergism with cyt1Aa: they are 1.5 to 34-fold more toxic than the respective clones without cyt1Aa against exposed larvae. Their lethal times also decreased (they kill larvae quicker), more so at higher cell concentrations. These clones are anticipated to dramatically reduce the likelihood of resistant development in the target organisms (Wirth et al., 1997).