Abstract
With the growth in the transgenic market, fast and economically viable methodologies are necessary for undertaking transgene detection tests, both for identification of contamination in seeds and in grain. Seeds from commercial conventional GNZ 2004, and transgenic VT-Pro (MON89034), Roundup Ready (NK603) and Herculex (TC1507) maize cultivars were used. In order to simulate different levels of contamination, the transgenic seeds were mixed with conventional seeds at levels of 0.2%, 0.4%, 1.0% and 1.6% for VT-Pro, and 0.2%, 0.5%, 0.8% and 1.2% for Roundup Ready and Herculex. The lateral flow membrane strip test was performed in the whole seed, endosperm and embryo. For evaluation of the specificity of the technique in detection of the TC1507 event by means of the conventional PCR technique, seeds of the commercial maize hybrid GNZ 2004 were used as negative control, and the maize hybrid 2B655Hx as positive control. In order to simulate different levels of contamination, transgenic seeds were mixed with conventional seeds at the levels of 10%, 5%, 1%, 0.5% and 0.1%. Seeds from each sample were crushed, and then DNA extraction was performed by the CTAB 2% method. Using the immunochromatographic strip, it was possible to evaluate the expression of proteins related to the VT-Pro, Roundup Ready and Herculex events when whole seeds were used at the 0.2% level of contamination, whereas by the conventional PCR technique, it was possible to detect the TC1507 event in samples with 1% contamination.
Highlights
In 2012, 170.3 million hectares of transgenic crops were grown in the world, with a growth of 10.3 million hectares in relation to the year 2011
This study was performed for the purpose of evaluating detection of protein expression of the VT-Pro event in maize grain lots containing different levels of contamination for the RR and Herculex events in the endosperm, in the embryo and in the whole seed, e and verifying which tissue is most recommended for performing the immunochromatographic strip test
It may be verified that the transgene was inserted in the female parent, seeing as how the combination of the quantity of proteins produced by the embryo and by the endosperm in the whole seed was necessary for there to be detection of the event at levels below that suggested by the company, and that detection in the embryo was only possible in a quantity of contamination greater than that recommended by the company
Summary
In 2012, 170.3 million hectares of transgenic crops were grown in the world, with a growth of 10.3 million hectares in relation to the year 2011. Tests based on proteins are not recommended in cases in which the protein, for some reason, is not being expressed in the plant tissue but the exogenous gene is present in its genome In these cases in which there is the need for directly analyzing the inserted gene, PCR has been the technique most used among all the DNA-based techniques (MARMIROLI et al, 2008). This study was performed for the purpose of evaluating detection of protein expression of the VT-Pro event in maize grain lots containing different levels of contamination for the RR and Herculex events in the endosperm, in the embryo and in the whole seed, e and verifying which tissue is most recommended for performing the immunochromatographic strip test. The purpose was to verify the sensitivity of the primer for identification of the TC1507 event in maize seeds, at different levels of contamination, using conventional PCR
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