Multiplex PCR-Based Simultaneous Amplification of Selectable Marker and Reporter Genes for the Screening of Genetically Modified Crops
- 27 May 2009
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of Agricultural and Food Chemistry
- Vol. 57 (12), 5167-5172
- https://doi.org/10.1021/jf900604h
Abstract
The development and commercialization of genetically modified (GM) crops with enhanced insect and herbicide resistance, abiotic stress tolerance, and improved nutritional quality has expanded dramatically. Notwithstanding the huge potential benefits of GM crops, the perceived environmental risks associated with these crops need to be addressed in proper perspective. One critical concern is the adventitious presence or unintentional mixing of GM seed in non-GM seed lots, which can seriously affect the global seed market. It would therefore be necessary though a challenging task to develop reliable, efficient, and economical assays for GM detection, identification, and quantification in non-GM seed lots. This can be systematically undertaken by preliminary screening for control elements and selectable or scorable (reporter) marker genes. In this study, simplex and multiplex polymerase chain reaction (PCR) assays individually as well as simultaneously amplifying the commonly used selectable marker genes, i.e., aadA, bar, hpt, nptII, pat encoding, respectively, for aminoglycoside-3′-adenyltransferase, Streptococcus viridochromogenes phosphinothricin-N-acetyltransferase, hygromycin phosphotransferase, neomycin phosphotransferase, Streptococcus hygroscopicus phosphinothricin-N-acetyltransferase, and a reporter gene uidA encoding β-d-glucuronidase, were developed as a reliable tool for qualitative screening of GM crops. The efficiency of the assays was also standardized in the test samples prepared by artificial mixing of transgenic seed samples in different proportions. The developed multiplex PCR assays will be useful in verifying the GM status of a sample irrespective of the crop and GM trait.Keywords
This publication has 11 references indexed in Scilit:
- Detection and characterization of recombinant DNA expressing vip3A-type insecticidal gene in GMOs—standard single, multiplex and construct-specific PCR assaysAnalytical and Bioanalytical Chemistry, 2007
- Detection of Roundup Ready soy in highly processed products by triplex nested PCRFood Control, 2007
- Genetically modified crops: success, safety assessment, and public concernApplied Microbiology and Biotechnology, 2006
- Development of a Seven-Target Multiplex PCR for the Simultaneous Detection of Transgenic Soybean and Maize in Feeds and FoodsJournal of Agricultural and Food Chemistry, 2004
- Validation of a Rice Specific Gene, Sucrose Phosphate Synthase, Used as the Endogenous Reference Gene for Qualitative and Real-Time Quantitative PCR Detection of TransgenesJournal of Agricultural and Food Chemistry, 2004
- Reliable Detection and Identification of Genetically Modified Maize, Soybean, and Canola by Multiplex PCR AnalysisJournal of Agricultural and Food Chemistry, 2003
- Screening of genetically modified organisms and specific detection of Bt176 maize in flours and starches by PCR-enzyme linked immunosorbent assayZeitschrift für Lebensmittel-Untersuchung und Forschung, 2003
- A Multiplex PCR Method of Detecting Recombinant DNAs from Five Lines of Genetically Modified Maize.Food Hygiene and Safety Science (Shokuhin Eiseigaku Zasshi), 2001
- Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplificationNucleic Acids Research, 1988
- A plant DNA minipreparation: Version IIPlant Molecular Biology Reporter, 1983