Development of an event-specific Real-time PCR detection method for the transgenic Bt rice line KMD1

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

doi:10.1007/s00217-008-0981-0

Cited by 44 publications

(21 citation statements)

References 50 publications

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“…However, Babekova et al [3] reported difficulties by applying the method on rice DNA resulting in amplicons of an average length of approximately 0.3 kb. Similar limitations occurred during application of the original protocol [2] on genomic DNA of Rhodococcus opacus 1CP yielding PCR products in a range of 0.1 -0.2 kb (data not shown).…”

Section: Resultsmentioning

confidence: 98%

“…One of these is SiteFinding-PCR which has been introduced as a simple and reliable PCR method for genome walking [2]. Since its first report in 2005 the method has been applied on genomic DNA from transgenic rice [3], the cyanophage P4 [2], and Pseudomonas putida JD1 [4] as well as on metagenomic samples from a bioleaching reactor [5]. However, to our knowledge DNA from Gram-positive bacteria and, in particular DNA with high GC content (e.g.…”

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confidence: 98%

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Optimization of a genome‐walking method to suit GC‐rich template DNA from biotechnological relevant Actinobacteria

Gröning¹,

Tischler²,

Kaschabek³

et al. 2010

J. Basic Microbiol.

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SiteFinding-PCR has been recently reported to be a useful technique in order to identify unknown DNA fragments located adjacent to available sequences. However, this method has so far only been applied to few DNA sources including plants, samples from bioleaching communities, and a Pseudomonas strain. In order to complete the sequence information of two gene clusters in Gram-positive rhodococci the original protocol was applied yielding amplicons of insufficient size. The binding site of the previously published SiteFinder-2 oligo proved to be unsuitable for Rhodococcus and other members of the Actinobacteria since the binding motif occurred too frequently. Available genome sequences of different Actinobacteria were analysed and the binding site of the SiteFinder oligo modified. Moreover, PCR conditions were adapted to the high GC content of the template DNA allowing the successful adaptation of this method to two members of the Actinobacteria.

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Section: Resultsmentioning

confidence: 98%

mentioning

confidence: 98%

Optimization of a genome‐walking method to suit GC‐rich template DNA from biotechnological relevant Actinobacteria

Gröning¹,

Tischler²,

Kaschabek³

et al. 2010

J. Basic Microbiol.

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“…EU880444.1), KMD1 (Accession no. EU980363.1) (Babekova et al 2009), KF6 (Accession no. HM124449) (Wang et al 2011).…”

Section: Pcr Primersmentioning

confidence: 99%

“…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%

“…Real-time PCR assays and loop-mediated isothermal amplification (LAMP) assays for KMD1, TT51-1 and KF6 have been reported (Babekova et al 2009;Wu et al 2010;Cao et al 2011;Chen et al 2012). However, multiplexing can provide a cost effective diagnostic assay for GM detection with higher throughput and less consumption of samples and reagents as compared to simplex assays (Bahrdt et al 2010;Kim et al 2013;Kim et al 2014).…”

Section: Introductionmentioning

confidence: 99%

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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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GMO Analysis Methods for Food: From Today to Tomorrow

2016

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Development of an event-specific Real-time PCR detection method for the transgenic Bt rice line KMD1

Cited by 44 publications

References 50 publications

Optimization of a genome‐walking method to suit GC‐rich template DNA from biotechnological relevant Actinobacteria

Optimization of a genome‐walking method to suit GC‐rich template DNA from biotechnological relevant Actinobacteria

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

GMO Analysis Methods for Food: From Today to Tomorrow

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