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Screening For Genetically Modified Tomatoes &amp / Tomato Seeds And Identification Of Cry1ac And Sam-k Specific Modifications Using Gene And Construct Specific Pcr

This study was carried out to analyze tomato samples and tomato seeds, purchased
from different food markets of Turkey randomly, for the presence of genetic
modification by using PCR method as it allows more specific detection. The DNAs
of collected samples were isolated according to CTAB DNA extraction protocol and
also with extraction kits. Screening tests of tomatoes were done by targeting 35S
promoter, NOS terminator and NptII kanamycin resistance gene with eight different
primer sets. Real time PCR is used to confirm 35S and NOS positives results
obtained from conventional PCR.
In this study, it was observed that 14 out of 35 seed samples, and 14 out of 40 fresh
tomato samples which were screened had at least one transgenic element of 35S promoter, NOS terminator and NPTII kanamycin resistance gene indicating the
possible presence of genetic modifications.
After screening, gene specific studies were carried out for PG, sam-k indicating F
type ripening delayed tomato and the 35 1 N lines respectively and cry1Ac genes
inserted in 5345-1 insect resistant tomato line. PG and sam-k specific primers were
not amplified in any of the samples investigated whereas 18 out of 75 samples were
cry1Ac positive and 1 out of 75 samples was sam-k positive. Positives were
confirmed by sequence analysis.
Additionally, construct specific primers specific to 5345-1 and 35 1 N lines were
designed. PCR amplicons indicate the existence of the construct sequence. In order
to verify the results, PCR products were sent to sequence analysis

Identiferoai:union.ndltd.org:METU/oai:etd.lib.metu.edu.tr:http://etd.lib.metu.edu.tr/upload/3/12608839/index.pdf
Date01 September 2007
CreatorsUckun, Esra
ContributorsGurakan, Candan Guzin
PublisherMETU
Source SetsMiddle East Technical Univ.
LanguageEnglish
Detected LanguageEnglish
TypeM.S. Thesis
Formattext/pdf
RightsTo liberate the content for public access

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