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Development of Taxon-Specific Sequences of Common Wheat for the Detection of Genetically Modified Wheat
- Source :
- Journal of Agricultural and Food Chemistry. 53:6294-6300
- Publication Year :
- 2005
- Publisher :
- American Chemical Society (ACS), 2005.
-
Abstract
- Qualitative and quantitative Polymerase Chain Reaction (PCR) systems aimed at the specific detection and quantification of common wheat DNA are described. Many countries have issued regulations to label foods that include genetically modified organisms (GMOs). PCR technology is widely recognized as a reliable and useful technique for the qualitative and quantitative detection of GMOs. Detection methods are needed to amplify a target GM gene, and the amplified results should be compared with those of the corresponding taxon-specific reference gene to obtain reliable results. This paper describes the development of a specific DNA sequence in the waxy-D1 gene for common wheat (Triticum aestivum L.) and the design of a specific primer pair and TaqMan probe on the waxy-D1 gene for PCR analysis. The primers amplified a product (Wx012) of 102 bp. It is indicated that the Wx012 DNA sequence is specific to common wheat, showing homogeneity in qualitative PCR results and very similar quantification accuracy along 19 distantly related common wheat varieties. In Southern blot and real-time PCR analyses, this sequence showed either a single or a low number of copy genes. In addition, by qualitative and quantitative PCR using wx012 primers and a wx012-T probe, the limits of detection of the common wheat genome were found to be about 15 copies, and the reproducibility was reliable. In consequence, the PCR system using wx012 primers and wx012-T probe is considered to be suitable for use as a common wheat-specific taxon-specific reference gene in DNA analyses, including GMO tests.
- Subjects :
- DNA, Plant
Sequence analysis
Molecular Sequence Data
Computational biology
Biology
Genetically modified wheat
Polymerase Chain Reaction
Sensitivity and Specificity
DNA sequencing
law.invention
Species Specificity
law
TaqMan
Common wheat
Triticum
Polymerase chain reaction
Southern blot
Base Sequence
Reproducibility of Results
food and beverages
Sequence Analysis, DNA
General Chemistry
Plants, Genetically Modified
Blotting, Southern
Real-time polymerase chain reaction
Seeds
General Agricultural and Biological Sciences
Subjects
Details
- ISSN :
- 15205118 and 00218561
- Volume :
- 53
- Database :
- OpenAIRE
- Journal :
- Journal of Agricultural and Food Chemistry
- Accession number :
- edsair.doi.dedup.....814a69e83a73a8f4f0db4faa124b87d1