Validity assessment of the detection method of maize event Bt10 through investigation of its molecular structure.
In March 2005, U.S. authorities informed the European Commission of the inadvertent release of unauthorized maize GM event Bt10 in their market and subsequently the grain channel. In the United States measures were taken to eliminate Bt10 from seed and grain supplies; in the European Union an embargo for maize gluten and brewer's grain import was implemented unless certified of Bt10 absence with a Bt10-specific PCR detection method. With the aim of assessing the validity of the Bt10 detection method, an in-depth analysis of the molecular organization of the genetic modification of this event was carried out by both the company Syngenta, who produced the event, and the European Commission Joint Research Centre, who validated the detection method. Using a variety of molecular analytical tools, both organizations found the genetic modification of event Bt10 to be very complex in structure, with rearrangements, inversions, and multiple copies of the structural elements (cry1Ab, pat, and the amp gene), interspersed with small genomic maize fragments. Southern blot analyses demonstrated that all Bt10 elements were found tightly linked on one large fragment, including the region that would generate the event-specific PCR amplicon of the Bt10 detection method. This study proposes a hypothetical map of the insert of event Bt10 and concludes that the validated detection method for event Bt10 is fit for its purpose.
European Commission, DG Joint Research Centre, Institute for Health and Consumer Protection, Ispra, Italy. Anne.milcamps@jrc.it
Anne
A
Milcamps
Scott
S
Rabe
Rebecca
R
Cade
Anic J
AJ
De Framond
Peter
P
Henriksson
Vance
V
Kramer
Duarte
D
Lisboa
Susana
S
Pastor-Benito
Michael G
MG
Willits
David
D
Lawrence
Guy
G
Van den Eede
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics*; D015139:Blotting, Southern; D018744:DNA, Plant / Q000032:analysis*; D004274:DNA, Recombinant / Q000032:analysis*; D004731:Endotoxins / Q000032:analysis / Q000235:genetics*; D005060:Europe; D015723:Gene Library; D006460:Hemolysin Proteins / Q000032:analysis / Q000235:genetics*; D007881:Legislation, Food; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012639:Seeds / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014481:United States; D003313:Zea mays / Q000235:genetics*
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D018744:DNA, Plant; D004274:DNA, Recombinant; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis