Biosensor technology and surface plasmon resonance for real-time detection of genetically modified Roundup Ready soybean gene sequences.
Biospecific interaction analysis (BIA) was performed using surface plasmon resonance (SPR) and biosensor technologies to detect genetically modified Roundup Ready soybean gene sequences. We first immobilized, on SA sensor chips, single-stranded biotinylated oligonucleotides containing soybean lectin and Roundup Ready gene sequences, and the efficiency of hybridization to oligonucleotide probes differing in length was determined. Second, we immobilized biotinylated PCR products from nontransgenic soybeans (genomes carrying only the lectin gene), as well as from genetically modified Roundup Ready soybean, and we injected the oligonucleotide probes. Furthermore, we used the sensor chips carrying either lectin and Roundup Ready soybean PCR products or 21-mer oligonucleotide as probes, and we injected both nonpurified and purified asymmetric PCR products. The results obtained show that 13 and 15 mer oligonucleotides are suitable probes to detect genetically modified Roundup Ready soybean gene sequences (either target oligonucleotides or PCR products) under standard BIA experimental conditions. By contrast, when 11 mer DNA probes were employed, no efficient hybridization was obtained. All the SPR-based formats were found to be useful for detection of Roundup Ready gene sequences, suggesting that these procedures are useful for the real-time monitoring of hybridization between target single-stranded PCR products, obtained by using as substrates DNA isolated from normal or transgenic soybeans, and oligonucleotide or PCR-generated probes, therefore enabling a one-step, nonradioactive protocol to perform detection.
Feriotto
Giordana
G
Biotechnology Center, Ferrara University, Via L. Borsari n. 46, 44100 Ferrara, Italy.
Borgatti
Monica
M
Mischiati
Carlo
C
Bianchi
Nicoletta
N
Gambari
Roberto
R
D015374:Biosensing Techniques; D001709:Biotechnology / Q000379:methods; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000235:genetics*; D020349:Surface Plasmon Resonance / Q000379:methods