Detection of Genetically Modified Soybean Using Peptide Nucleic Acids (PNAs) and Microarray Technology

Peptide nucleic acid (PNA) microarrays for the detection of Roundup Ready soybeans in food have been prepared. PNA probes are known to be more efficient and selective in binding DNA sequences than the analogous oligonucleotides and are very suitable to be used for diagnostics in food. PNAs of differ...

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Bibliographic Details
Published in:Journal of agricultural and food chemistry 2004-07, Vol.52 (14), p.4535-4540
Main Authors: Germini, Andrea, Mezzelani, Alessandra, Lesignoli, Francesca, Corradini, Roberto, Marchelli, Rosangela, Bordoni, Roberta, Consolandi, Clarissa, De Bellis, Gianluca
Format: Article
Language:English
Subjects:
5-enolpyruvylshikimate-3-phosphate synthase
alkyl (aryl) transferases
Biological and medical sciences
DNA probes
DNA, Plant - analysis
food analysis
Food industries
Fruit and vegetable industries
Fundamental and applied biological sciences. Psychology
General aspects
genetically modified foods
Glycine max
Glycine max - genetics
glyphosate
herbicide resistance
Methods of analysis, processing and quality control, regulation, standards
microarray technology
nucleic acids
Oligonucleotide Array Sequence Analysis
Peptide Nucleic Acids - analysis
peptides
Plants, Genetically Modified - genetics
Polymerase Chain Reaction
Roundup Ready soybean
soy flour
soybeans
transgenes
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Summary:Peptide nucleic acid (PNA) microarrays for the detection of Roundup Ready soybeans in food have been prepared. PNA probes are known to be more efficient and selective in binding DNA sequences than the analogous oligonucleotides and are very suitable to be used for diagnostics in food. PNAs of different lengths were carefully designed and synthesized by solid-phase synthesis on an automatic synthesizer adopting the BOC strategy. PNAs were purified by HPLC and characterized by HPLC/MS. The probes were spotted on a functionalized surface to produce a microarray to be hybridized with PCR products. DNA extracted from reference material was amplified using Cy3- and Cy5-labeled primers, and the fluorescent PCR products obtained were hybridized on the microarray. Two protocols were adopted:  the hybridization with dsDNA or with ssDNA obtained by digestion with the enzyme λ exonuclease. The best results were obtained using a 15-mer PNA probe in combination with the ssPCR product derived from enzymatic digestion. The method was applied to the analysis of a sample of certified transgenic soybean flour. Keywords: GMO; PNA; microarrays; Roundup Ready soybean
ISSN:0021-8561
1520-5118
DOI:10.1021/jf035355r