Duplex Polymerase Chain Reaction (PCR) for the Simultaneous Detection of<i>Cryia(B)</i>and the Maize Ubiquitin Promoter in the Transgenic Rice Line Kmd1

Babekova, R.; Funk, Tristan; Pecoraro, Sven; Engel, Karl‐Heinz; Baikova, D.; Busch, Ulrich

doi:10.1080/13102818.2008.10817538

Cited by 8 publications

(3 citation statements)

References 16 publications

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“…Detection methods for GM rice line 'Kemingdao' ('KMD1') expressing the cryIA(b) gene driven by a maize ubiquitin promoter were published recently (Babekova et al 2008(Babekova et al , 2009). This line is a progeny of the commercial japonica rice variety Xiushui 11 genetically transformed with a cryIA(b) gene using the binary vector pKUB (Cheng et al 1998;Wu et al 2002).…”

mentioning

confidence: 99%

A testing cascade for the detection of genetically modified rice by real-time PCR in food and its application for detection of an unauthorized rice line similar to KeFeng6

2010

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mentioning

confidence: 99%

A testing cascade for the detection of genetically modified rice by real-time PCR in food and its application for detection of an unauthorized rice line similar to KeFeng6

2010

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“…Sebagai alternatif, teknik PCR secara multipleks dapat digunakan dalam meningkatkan efisiensi teknik deteksi PRG yang mengandung lebih dari satu gen interes dalam suatu genom atau multi event dalam suatu sampel, baik tanaman, makanan, maupun pakan ternak, secara simultan. Teknik PCR secara dupleks atau multipleks telah digunakan oleh beberapa peneliti untuk deteksi PRG baik yang berupa event tunggal atau stacked (Babekova et al, 2008;Hedreyda dan Roxas, 2010;Xu et al, 2009). Namun demikian, teknik PCR secara multipleks biasanya memerlukan optimasi kondisi PCR untuk mengatasi rendahnya sensitivitas produk amplifikasi atau amplikon yang dihasilkan oleh primer yang berbedabeda.…”

Section: Hasil Dan Pembahasanunclassified

Teknik PCR Kualitatif untuk Deteksi Produk Rekayasa Genetika Jagung Event BT11 dan GA21

Bahagiawati¹,

Reflinur²,

Santoso³

2015

J. AgroBiogen

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In some countries, including Indonesia, labelling of GMO products is mandatory for giving consumers the right to choose between GMOs and conventional products. Therefore, development of methodology that can detect a specific genetically modified (GM) crops and to verify the absence or presence of GM material in a product including raw materials (e.g. grains) and/or their derivatives is needed. The objectives of this study were to find the most efficient screening methods to detect whether or not a product is GM material and to develop a specific detection method to identify GM product BT11 and GA21. In addition, present study was also aimed to obtain a duplex detection method for both GM products. Two GM-maize, including the BT11 and GA21 lines of maize (Zea mays L.), and one plant, namely NK11 as the nontransgenic control, were used as plant genetic materials in the event-specific detection of maize. The target gene from each sample was amplified in different reaction (simplex) using both the event specific primer and the endogenous maize reference, Zein, as internal control. Furthermore, in duplex PCR, two targets were simultaneously amplified in the same reaction. The results showed that detection method of the GM product obtained from present study enabled us to screen the GM products and specifically the event of BT11 and GA21 using simplex and duplex methods. The duplex method is more efficient because it can detect two GM crops in one time compared to simplex method that only can detect GM crop one by one.Keywords: GMO detection, GMO screening, maize, BT11, GA21. ABSTRAKPelabelan produk hasil rekayasa genetika (PRG) di beberapa negara, termasuk Indonesia, merupakan mandat yang memberikan kesempatan kepada konsumen untuk mendapatkan informasi produk yang akan dibeli atau digunakan. Untuk itu, diperlukan perangkat teknologi yang dapat mendeteksi keberadaan produk PRG tersebut, baik dalam bentuk bahan dasar (bijibijian) maupun produk turunannya. Penelitian ini bertujuan membandingkan dan mendapatkan teknik yang akurat dan efisien untuk deteksi tanaman jagung BT11 dan GA21 berdasarkan teknik PCR, baik secara tunggal (simpleks) maupun ganda (dupleks). Jagung produk PRG, yaitu BT11 dan GA21, dan non-PRG, yaitu Nk11, digunakan sebagai sampel dalam pengembangan metode deteksi PRG pada kedua event tersebut. Setiap sampel diamplifikasi secara simpleks menggunakan primer spesifik untuk mendeteksi keberadaan gen target pada event PRG dan sepasang primer spesifik sebagai internal kontrol untuk tanaman jagung. Selanjutnya, pada metode dupleks, sampel DNA kedua event diamplifikasi dengan primer-primer tersebut secara bersamaan dalam suatu reaksi PCR. Hasil penelitian menunjukkan bahwa teknik deteksi yang didapatkan dari penelitian ini dapat digunakan untuk skrining PRG dan identifikasi spesifik BT11 dan GA21, baik secara simpleks maupun dupleks. Kemampuan metode dupleks untuk mendeteksi PRG akan sangat bermanfaat dalam kegiatan skrining PRG dalam jumlah sampel yang besar karena metode ini lebih efisien dibanding...

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“…TT51-1 is an insect-resistant transgenic rice event harboring a hybrid cry1Ab/Ac gene driven by rice actin 1 gene promoter and the nopaline synthase (NOS) terminator, which was granted the safety certificate by China in 2009 (Tu et al 2003;Lu 2010). The transgenic Chinese Bt rice KMD1, derived from the commercial japonica rice variety Xiushui11 (XS11) by Agrobacterium transformation with a synthetic cry1Ab gene (Babekova et al 2008;Liu et al 2012), is resistant to eight lepidopteran pest species like the yellow stem borer, the striped stem borer and the rice leaf folder, which has the potential to be approved in China (Babekova et al 2009). KF6 contains two insect-resistant genes, cry1Ac gene under the control of the maize ubiqutin promoter and CpTI (cowpea trypsin inhibitor) gene (Rong et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Development of a multiplex event-specific PCR assay for detection of genetically modified rice

Lü

et al. 2016

Cereal Research Communications

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Global rice supplies have been found contaminated with unapproved varieties of genetically modified (GM) rice in recent years, which has led to product recalls in several of countries. Faster and more effective detection of GM contamination can prevent adulterated food, feed and seed from being consumed and grown, minimize the potential environmental, health or economic damage. In this study, a simple, reliable and cost-effective multiplex polymerase chain reaction (PCR) assay for identifying genetic modifications of TT51-1, Kemingdao1 (KMD1) and Kefeng6 (KF6) rice was developed by using the event-specific fragment. The limit of detection (LOD) for each event in the multiplex PCR is approximately 0.1%. Developed multiplex PCR assays can provide a rapid and simultaneous detection of GM rice.

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Duplex Polymerase Chain Reaction (PCR) for the Simultaneous Detection ofCryia(B)and the Maize Ubiquitin Promoter in the Transgenic Rice Line Kmd1

Cited by 8 publications

References 16 publications

A testing cascade for the detection of genetically modified rice by real-time PCR in food and its application for detection of an unauthorized rice line similar to KeFeng6

A testing cascade for the detection of genetically modified rice by real-time PCR in food and its application for detection of an unauthorized rice line similar to KeFeng6

Teknik PCR Kualitatif untuk Deteksi Produk Rekayasa Genetika Jagung Event BT11 dan GA21

Development of a multiplex event-specific PCR assay for detection of genetically modified rice

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