use of 35S and Tnos expression elements in the measurement of genetically engineered plant materials

• Published in: Analytical and bioanalytical chemistry 2010-03, Vol.396 (6), p.2175-2187
• Main Authors: Holden, Marcia J, Levine, Marci, Scholdberg, Tandace, Haynes, Ross J, Jenkins, G. Ronald
• Format: Article
• Language: English
• Subjects: Agrobacterium tumefaciens - genetics; Analysis; Analytical Chemistry; Base Sequence; Biochemistry; Biotechnology; Caulimovirus - genetics; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Corn; Food; Food Science; Gel electrophoresis; Gene amplification; Genetic aspects; Genetic Engineering; Genetic Vectors - genetics; Genetically engineered foods; Genetically modified crops; Identification and classification; Laboratory Medicine; Laws, regulations and rules; Mathematical analysis; Methods; Molecular Sequence Data; Monitoring/Environmental Analysis; Nucleic acids; Nucleotide sequence; Original Paper; Physiological aspects; Plants (organisms); Plants, Genetically Modified - genetics; Plants, Genetically Modified - microbiology; Plants, Genetically Modified - virology; Polyadenylation; Polymerase chain reaction; Polymerase Chain Reaction - methods; Primers (Molecular genetics); Promoter Regions, Genetic; Promoters (Genetics); Scientific surveys; Separation; Zea mays - genetics; Zea mays - microbiology; Zea mays - virology
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-009-3186-x

An online survey was conducted by the International Life Sciences Institute, Food Biotechnology Committee, on the use of qualitative and quantitative polymerase chain reaction (PCR) assays for cauliflower mosaic virus 35S promoter and Agrobacterium tumefaciens Tnos DNA sequence elements for the detection of genetically engineered (GE) crop plant material. Forty-four testing laboratories around the world completed the survey. The results showed the widespread use of such methods, the multiplicity of published and in-house methods, and the variety of reference materials and calibrants in use. There was an interest on the part of respondents in validated quantitative assays relevant to all GE events that contain these two genetic elements. Data are presented by testing two variations each of five published real-time quantitative PCR methods for 35S detection on eight maize reference materials. The results showed that two of the five methods were not suitable for all the eight reference materials, with poor linear regression parameters and multiple PCR amplification products for some of the reference materials. This study demonstrates that not all 35S methods produce satisfactory results, emphasizing the need for method validation.