Investigating The GMO Existence in Chips and Breakfast Cereals Marketed in Turkey

Yıl 2021, , 375 - 385, 07.09.2021

Şebnem Mutlu , Osman Şimşek , Ömer Öksüz

https://doi.org/10.33462/jotaf.670008

Öz

In this research, processed or low processed samples containing corn or corn products (corn semolina, flour, etc.) and soybean were randomly collected from the market, and 25 products in total (chips, nuts, cereals, flour) were analyzed for genetic modification using DNA based detection method, the polymerase chain reaction. First, homogenization of the samples was performed. Then DNA isolation was done by using Cetyl Trimethyl Ammonium Bromide (CTAB) and Roche High Pure DNA Isolation Kit. Since the Roche High Pure DNA Isolation Kit gave better results, the analysis was completed with this method. After DNA isolation, the detection of the Lectin gene, Zein gene, CaMV 35S Promoter and NOS Terminator regions was performed by conventional PCR. Zein gene determination was done for searching and proving corn presence and similarly, Lectin gene determination was done for searching and proving soybean presence in the samples by conventional PCR. GMO3/GMO4 and Zein3/Zein4 primer pairs were used for Lectin and Zein gene determination, respectively. The amplification of DNA was observed in agarose gel electrophoresis. Lectin or Zein genes were detected in 17 samples while these genes were not detected in 8 samples. Samples, in which Lectin or Zein gene was detected were scanned for 35S promoter or NOS terminator. 35S-3/35S-6 and tNOS2F/tNOS2R primer pairs were used for scanning 35S Promoter and NOS Terminator, respectively. To observe possible contamination in the mix sterilized deionized water was used and 0% Bt-11 and 0% GTS 40-3-2 were used as a negative control, 5% Bt-11 and 10% GTS-40-3-2 were used as a positive control. All of the 25 samples did not provide enough DNA with the required quality. This result was considered to be sourced by the applications (frying, extruding, pressing etc.) that samples had been exposed to during processes. Neither 35S Promotor nor NOS Terminator was determined from any of the samples.

Anahtar Kelimeler

GMO, Convantional PCR, Lectin, Zein, 35S Promotor, NOS Terminator

Destekleyen Kurum

NKU Bilimsel Araştırma Projeleri Koordinasyon Birimi

Proje Numarası

NKUBAP.00.24.AR.13.13 .

Teşekkür

We thank to Namık Kemal University Scientific Research Projects Coordination Department (NKUBAP) for financial support. (Project Number NKUBAP.00.24.AR.13.13)

Investigating The GMO Existence in Chips and Breakfast Cereals Marketed in Turkey

Öz

Bu çalışmada piyasada satılan mısır ya da mısır ürünleri (mısır irmiği, mısır unu vb.) veya soya içeren işlenmiş veya yarı işlenmiş ürünler rastgele seçilerek toplamda 25 adet üründe (cips, çerez, gevrek, kuruyemiş kaplaması, un) GDO varlığı DNA temelli bir tespit metodu olan konvansiyonel Polimeraz Zincir Reaksiyonu (PCR) ile araştırılmıştır. Bu amaçla homojenize edilen örneklerde CTAB (Cetyl Trimethyl Ammonium Bromide) yöntemi ve Roche High Pure DNA Isolation Kit ile DNA izole edilerek çoğaltılması denenmiştir. DNA Isolation Kit kullanılarak DNA izole edilmesi yöntemi daha başarılı olduğundan işlemler bu yöntemle tamamlanmıştır. DNA izolasyonunun ardından Lektin geni, Zein geni, 35S Promotör ve NOS Terminatör bölgelerinin tespiti konvansiyonel PCR ile yapılmıştır. Zein geni tespiti örneklerdeki mısır varlığını, Lektin geni tespiti ise örneklerdeki soya varlığını araştırmak ve kanıtlamak amacıyla yapılmıştır. Zein ve Lektin genleri tespiti için sırasıyla GMO3/GMO4 ve Zein3/Zein4 primer çiftleri kullanılmıştır. Elde edilen DNA’ların amplifikasyonundan sonra DNA agaroz jel elektroforezi ile gözlemlenmiştir. Agaroz jel elektroforezi sonucu alınan görüntülerde 17 örnekte Lektin ya da Zein geni tespit edilirken 8 örnekte bu genler tespit edilememiştir. Lektin ya da Zein geni tespit edilen ürünlerin 35S Promotör ve NOS Terminatör taraması için çalışılmaya devam edilmiştir. Bunun için sırasıyla 35S-3/35S-6 ve tNOS2F/tNOS2R primer çiftleri kullanılmıştır. Mümkün kontaminasyonları gözlemlemek için sterilize su kullanılırken, negatif kontrol amacıyla 0 % Bt-11 ve 0% GTS 40-3-2, pozitif kontrol amacıyla ise 5% Bt-11 ve 10% GTS-40-3-2 kullanılmıştır. 25 örneğin hiçbirinde istenen kalitede ve yeterli miktarda DNA elde edilememesinin sebebinin işleme sırasında uygulanan prosesler (kızartma, kavurma, ekstrude etme, basınç vb.) olduğu düşünülmektedir. Hiçbir numunede 35S Promotör ve NOS Terminatör bölgesine rastlanmamıştır.

Anahtar Kelimeler

GDO, Konvansiyonel PCR, Lektin geni, Zein geni, 35S Promotör, NOS Terminatör

Proje Numarası

NKUBAP.00.24.AR.13.13 .

Kaynakça

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