Detection and identification of cry1I genes in Bacillus thuringiensis using polymerase chain reaction and restriction fragment length polymorphism analysis.
A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for detection and identification of cry1I genes from Bacillus thuringiensis (Bt) was established. Based on the analysis of conserved regions of the cry1I genes, 2 oligonucleotide primers were designed to amplify a 665-bp fragment of the genes. The amplification products were digested with restriction endonuclease HinfI or with RsaI in addition for specific detection of different variants from the known subclasses of cry1I genes. The PCR-RFLP pattern obtained revealed the detection of cry1I genes in 151 of 202 native Bt isolates. Furthermore, cry1I genes were detectable in 10 of 19 standard strains tested. The cry1Ia gene was the most abundant cry1I gene subclass present in 54 of 56 native Bt isolates and in 8 of 10 standard strains. Based on the results obtained, the PCR-RFLP method may be a valuable and reliable tool for specific detection and identification of cry1I genes.
Sauka
Diego H
DH
Area Bioinsumos Microbianos, Instituto de Microbiología y Zoología Agrícola, Instituto Nacional de Tecnología Agropecuaria, De Los Reseros y Las Cabañas s/nro., C.C. 25, C.P. 1712 Castelar, Buenos Aires, Argentina. dsauka@cnia.inta.gov.ar
Cozzi
Jorge G
JG
Benintende
Graciela B
GB
D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004731:Endotoxins / Q000235:genetics*; D006460:Hemolysin Proteins; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D017931:DNA Primers; D004269:DNA, Bacterial; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D015252:Deoxyribonucleases, Type II Site-Specific; C032229:GANTC-specific type II deoxyribonucleases; C040158:GTAC-specific type II deoxyribonucleases