dominant negative mutants of bacillus thuringiensis cry1ab toxin function as anti-toxins demonstration of the role of oligomerization in toxicity显性负突变体的苏云金杆菌cry1ab毒素作为抗示范齐聚在毒性的作用.pdfVIP

Dominant Negative Mutants of Bacillus thuringiensis Cry1Ab Toxin Function as Anti-Toxins: Demonstration of the Role of Oligomerization in Toxicity ´ ´ 1 1 ˜ 1 ´ ´ 1 Claudia Rodrıguez-Almazan , Luis Enrique Zavala , Carlos Munoz-Garay , Nuria Jimenez-Juarez , 1 2 ´ 1 1 Sabino Pacheco , Luke Masson , Mario Soberon , Alejandra Bravo * ´ ´ ´ 1 Instituto de Biotecnologıa, Universidad Nacional Autonoma de Mexico, Morelos, Mexico, 2 Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec, Canada Abstract Background: Bacillus thuringiensis Cry toxins, that are used worldwide in insect control, kill insects by a mechanism that depends on their ability to form oligomeric pores that insert into the insect-midgut cells. These toxins are being used worldwide in transgenic plants or spray to control insect pests in agriculture. However, a major concern has been the possible effects of these insecticidal proteins on non-target organisms mainly in ecosystems adjacent to agricultural fields. Methodology/Principal Findings: We isolated and characterized 11 non-toxic mutants of Cry1Ab toxin affected in different steps of the mechanism of action namely binding to receptors, oligomerization and pore-formation. These mutant toxins were analyzed for their capacity to block wild type toxin activity, presenting a dominant negative phenotype. The dominant negative phenotype was analyzed at two levels, in vivo by toxicity bioassays against susceptible Manduca sexta larvae and in vitro by pore formation activity in black lipid bi