Isolamento e caracterização molecular de Bacillus thuringiensis (Bt) na região oeste do Paraná

Abstract

The bacterium Bacillus thuringiensis (Bt), gram-positive and sporulant wand shaped, which can produce an attached crystal to the spore with toxicological potential. Due to this potential and extensively studied for use as a bioinsecticide against agricultural praises of Lepidoptera, Coleoptera and Diptera, and urban praises such as the Aedes aegypti gnat, this DNA can be used to develop transgenics. What makes the search for new haplotypes constant. Our goal was to search for new isolates, performing soil collections in areas of conventional cultivation, urban forest and legal reserve in western Paraná. Samples were isolated in ampicillin nutrient medium, colonies morphologically characterized, DNA extraction, PCR reaction for 16S, gyrB, cry1, 2, 3, 4, 10, 11 genes. For quantification and confirmation of all results were performed electrophoresis in agarose gel, followed by purification with polyethylene glycol and sent to sequencing by a third-party, when the band size is found like expected. The sequences were edited and compared to the GenBank sequence (https://www.ncbi.nlm.nih.gov/genbank/) for molecular identification. The 16S and gyrB results confirm that 16 isolates may be Bacillus thuringienses and it is possible that they may have the cry4 gene.