The Use of 35S and Tnos Expression Elements in the Measurement of Genetically Engineered Plant Materials
Holden MJ, Levine M, Scholdberg T, Haynes RJ, Jenkins GR
Analytical and Bioanalytical Chemistry
January 1, 2009
Sampling and Detection
Analytical and Bioanalytical Chemistry. 2010;396(6):2175-2187
Abstract: 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.
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- Lee L-Y, Gelvin SB. T-DNA binary vectors and systems. Plant Physiol. 2008;146:325–332. doi: 10.1104/pp.107.113001. LINK
- Plant Viruses Online. LINK. Accessed March 2009.
- Benfey PN, Chua N-H. The cauliflower mosaic virus 35S promoter: combinatorial regulation of transcription in plants. Science. 1990;250:959–966. doi: 10.1126/science.250.4983.959. LINK
- Agbios GM Database. LINK. Accessed June 4, 2009.
- Lipp M, Shillito R, Giroux R, Spiegelhalter F, Charlton S, Pinero D, Song P. Polymerase chain reaction technology as analytical tool in agricultural biotechnology. J AOAC Int. 2005;88:136–155. LINK
- Chaouachi M, Chupeau G, Berapd A, McKhann H, Romaniuk M, Giancola S, Daval V, Bertheau Y, Brunel D. J Agric Food Chem. 2008;56:11596–11606. doi: 10.1021/jf801482r. LINK
- Leimanis S, Hamels S, Naze F, Mbella MM, Sneyers M, Hochegger R, Broll H, Roth L, Dallmann K, Micsinal A, LaPaz JL, Pla M, Brunen-Nieweler C, Papazova N, Taverniers I, Hess N, Kirschneit B, Bertheau Y, Audeon C, Laval V, Busch U, Pecoraro S, Neumann K, Rosel S, VanDijk J. Kok Esther, Bellocchi G, Foti N, Mazzara M, Moens W, Remacle J, Van Den Eede G. Eur Food Res Technol. 2008;227:1621–1632. doi: 10.1007/s00217-008-0886-y. LINK
- Layton DT, Spieglehalter F, Jenkins R (2008) How grain companies are managing the challenges posed by stacked events in meeting the global regulatory and commercial requirements for non-GM corn shipments: a comparison of methods in current use. 1st Global Conference on GMO Analysis, Como Italy. LINK.
- Survey Monkey. LINK. Accessed June 22, 2009.
- European Commission, Joint Research Center, GMO methods database. LINK. Accessed June 22, 2009.
- Dong W, Yang L, Shen K, Banghyun K, Kleter GA, Marvin HJP, Guo R, Liang W, Zhang D. GMDD: a database of GMO detection methods. BMC Bioinformatics. 2008;9:260. doi: 10.1186/1471-2105-9-260. LINK
- Scholdberg TA, Norden TD, Nelson DD, Jenkins GR. Evaluating precision and accuracy when quantifying different endogenous control reference genes in maize using real-time PCR. J Agric Food Chem. 2009;57:2903–2911. doi: 10.1021/jf803599t. LINK
- European Commission, Joint Research Center, Community reference laboratory. Accessed June 22, 2009. LINK
- European Commission, Joint Research Center, Institute for Reference Materials and Measurements. LINK. Accessed June 22, 2009.
- American Oil Chemists Society. LINK.
- Wolf C, Scherzinger M, Wruz A, Pauli U, Hubner P, Luthy J. Detection of cauliflower mosaic virus by the polymerase chain reaction: testing of food components for false positive 35S-promoter screening results. Eur Food Res Technol. 2000;210:367–372. doi: 10.1007/s002170050565. LINK
- Chaouachi M, Fortabat MN, Geldreich A, Yot P, Kerlan C, Kevdani N, Audeon C, Romaniak M, Bertheau Y. An accurate real-time PCR test for the detection and quantification of cauliflower mosaic virus (CaMV): applicable in GMO screening. Eur Food Res Technol. 2008;227:789–798. doi: 10.1007/s00217-007-0787-5. LINK
- EC Joint Research Center, Community Reference Laboratory, DNA Extraction method.. Accessed May 12, 2009. LINK
- Arumuganathan K, Earle ED. Nuclear DNA content of some important plant species. J Agric Food Chem. 1991;9:208–218. LINK
- Rayburn AL, Biradar DP, Bullock DG, McMurphy LM. Nuclear DNA content in F1 hybrids of maize. Heredity. 1992;70:294–300. doi: 10.1038/hdy.1993.42. LINK
- Bennett MD, Leitch IJ (2004) Plant DNA C-values database (release 5.0, Dec. 2004). LINK. Accessed June 24, 2009.
- Kuribara H, Shindo Y, Matsuoka T, Takubo K, Futo S, Aoki N, Hirao T, Akiyama H, Goda Y, Toyada M, Hino A. Novel reference molecules for quantitation of genetically modified maize and soybean. J AOAC Int. 2002;85:1077–1089. LINK
- Corbisier P, Trapmann S, Gancberg D, Hannes L, Iwaarden P, Berben G, Schimmel H, Emons H. Quantitative determination of Roundup Ready soybean (Glycine max) extracted from highly processed flour. Anal Bioanal Chem. 2005;383:282–290. doi: 10.1007/s00216-005-0013-x. LINK
- Fernandez S, Charles-Delobel C, Geldreich A, Berthier G, Boyer F, Collonnier C, Coue-Philippe G, Diolez A, Duplan M-N, Kebdani N, Romaniuk M, Feinberg M, Bertheau Y. Quantification of the 35S promoter in DNA extracts from genetically modified organisms using real-time polymerase chain reaction and specificity assessment on various genetically modified organisms. Part 1: operating procedure. J AOAC Int. 2005;88:547–557. LINK
- Zeitler R, Pietsch K, Waiblinger H-U. Validation of real-time PCR methods for quantification of transgenic contamination in rape seed. Eur Food Res Technol. 2002;214:346–351. doi: 10.1007/s00217-001-0454-1. LINK
- Hohne M, Santisi CR, Meyer R. Real-time multiplex PCR: an accurate method for the detection and quantification of 35S-CaMV promoter in genetically modified maize-containing food. Eur Food Res Technol. 2002;215:59–64. doi: 10.1007/s00217-002-0503-4. LINK
- Pfaffl MW. In: A-Z of quantitative PCR. Bustin SA, editor. La Jolla: International University Line; 2004. pp. 87–112. LINK
- Brisson M, Hall S, Hamby RK, Park R, Srere HK. In: A-Z of quantitative PCR. Bustin SA, editor. La Jolla: International University Line; 2004. pp. 619–642. LINK
- Morisett D, Demsar T, Gruden K, Vojvoda J, Stebih D, Zel J. Detection of genetically modified organisms—closing the gaps. Nature Biotechnol. 2009;27:700–701. doi: 10.1038/nbt0809-700. LINK
- Feinberg M, Fernandez S, Cassard S, Charles-Delobel C, Bertheau Y. Quantitation of 35S promoter in maize DNA extracts from genetically modified organisms using real-time polymerase chain reaction, part 2: interlaboratory study. J AOAC Int. 2005;88:558–573. LINK