PCR analysis of the cryI insecticidal crystal family genes from Bacillus thuringiensis

• Published in: Applied and Environmental Microbiology Vol. 60; no. 1; pp. 353 - 356
• Main Authors: CERON, J, COVARRUBIAS, L, QUINTERO, R, ORTIZ, A, ORTIZ, M, ARANDA, E, LINA, L, BRAVO, A
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 1994
• Subjects: amplification chaine polymerase; Bacillus thuringiensis; Bacillus thuringiensis - classification; Bacillus thuringiensis - genetics; Bacillus thuringiensis - isolation & purification; Bacteria; bacterial pesticides; Bacterial Proteins - classification; Bacterial Proteins - genetics; Bacterial Toxins; Base Sequence; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; DNA, Bacterial - genetics; endotoxinas; endotoxine; endotoxins; Endotoxins - classification; Endotoxins - genetics; Evaluation Studies as Topic; Fundamental and applied biological sciences. Psychology; gene; Genes; Genes, Bacterial; Genetics; Hemolysin Proteins; insecticidas; insecticide; insecticides; Mexico; Mission oriented research; Molecular Sequence Data; Multigene Family; Pest Control, Biological; pesticide bacterien; plaguicidas bacterianos; polymerase chain reaction; Polymerase Chain Reaction - methods; Proteins; reaccion de cadenas de polimerasa; Soil Microbiology; toxinas; toxine; toxins; Mexico; Polymerase chain reaction; Toxin; Bacillales; Bacillus thuringiensis; Electrophoretic mobility; Reaction product; Bacteria; Multigene family; Subgroup; Identification; Bacillaceae; Microbial insecticide
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.60.1.353-356.1994

A method allowing rapid and accurate identification of different subgroups within the insecticidal crystal CryI protein-producing family of Bacillus thuringiensis strains was established by using PCR technology. Thirteen highly homologous primers specific to regions within genes encoding seven different subgroups of B. thuringiensis CryI proteins were described. Differentiation among these strains was determined on the basis of the electrophoretic patterns of PCR products. B. thuringiensis strains, isolated from soil samples, were analyzed by PCR technology. Small amounts of bacterial lysates were assayed in two reaction mixtures containing six to eight primers. This method can be applied to rapidly detect the subgroups of CryI proteins that correspond with toxicity to various lepidopteran insects.