RESUMO
The insecticidal crystal proteins of 15 B. thuringiensis strains, isolated in the Crimea territory that are toxical for some Lepidoptera and Colorado potato beetle larvae were identified by PAGE electrophoresis. Ten strains produced the crystal proteins with high molecular weight (> 120 kD). PCR with use of broad specificity primers and DNA of these B. thuringiensis strains as template demonstrated the specific PCR products (1000 bp). Amplified DNA fragments were cloned and sequenced. The nucleotide sequence analysis revealed that the genomes of ten strains of B. thuringiensis carried Cry1B genes, which are responsible for production of the insecticidal crystal proteins with dual specificity. The influence of the solubilization conditions on the structure and toxicity of Cry1B protein for Colorado potato beetle larvae was shown. The dual toxicity of studied B. thuringiensis strains is explained by the Cry1B genes presence in their genomes. These strains may be used to develop the broad specificity bioinsecticides.
Assuntos
Bacillus thuringiensis/genética , Proteínas de Bactérias , Toxinas Bacterianas , Endotoxinas , Genes Bacterianos , Proteínas Hemolisinas , Microbiologia do Solo , Animais , Bacillus thuringiensis/isolamento & purificação , Toxinas de Bacillus thuringiensis , Proteínas de Bactérias/genética , Proteínas de Bactérias/farmacologia , Proteínas de Bactérias/fisiologia , Toxinas Bacterianas/genética , Toxinas Bacterianas/farmacologia , Besouros/efeitos dos fármacos , Besouros/crescimento & desenvolvimento , DNA Bacteriano , Eletroforese em Gel de Poliacrilamida , Endotoxinas/genética , Endotoxinas/farmacologia , Endotoxinas/fisiologia , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/farmacologia , Proteínas Hemolisinas/fisiologia , Larva/efeitos dos fármacos , Plasmídeos , UcrâniaRESUMO
The specific to 3 types of Cry genes primers containing inosine were constructed to find crystal insecticidal protein Bacillus thuringiensis genes using PCR. A number of new B. thuringiensis strains isolated in Ukraine were investigated using these PCR primers. As a results, Cry genes were found, some of them were identified and demonstrated high homology to Cry1Ba2 and Cry1Bc genes.
