Identity tags revisited

Marillonnet, Sylvestre; Klimyuk, Victor; Gleba, Yuri

doi:10.1038/nbt0903-974

Nat Biotechnol

2003

DOI: 10.1038/nbt0903-974

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Identity tags revisited

Sylvestre Marillonnet¹,

Victor Klimyuk²,

Yuri Gleba³

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“…However, as argued by Pauli and Marillonnet et al (23,26), there are still some points to be considered, such as the need for extensive verification of safety, the requirement for additional sequencing, and most importantly, gene-crossing problems caused by horizontal gene transfer (HGT), leading to the separation of tag and transgene. The gene-crossing problems are more serious in GMMs, since gene transfer is suspected to be a common event in microbial communities (10,21).…”

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confidence: 99%

“Mark the Gene”: a Method for Nondestructive Introduction of Marker Sequences Inside the Gene Frame of Transgenes

Morono

Kitagawa

Kimura

et al. 2007

Appl Environ Microbiol

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A specific marking and detection technique is a fundamental requirement for the safer use of genetically modified (GM) organisms. Here we propose a simple and effective method for directly marking functional transgenes in GM organisms. For that purpose, we introduced nucleotide substitutions (NS), based on the degeneracy of codons as markers (NS markers), into the bphC (2,3-dihydroxybiphenyl dioxygenase) and tomA3 (toluene-ortho-monooxygenase) gene frames using a PCR-based method. No change was observed in the enzyme activity of translated proteins, and alignments with homologous genes showed the uniqueness of the NS markers. Furthermore, we constructed tomA3 variations harboring NS markers in different positions. Although the translational products were identical, the constructed variation genes could be distinguished through their marker patterns by multiplex PCR, showing that NS markers could serve as product-specific tags for identifying individual GM organisms. This direct method of marking the functional transgene provides a simple, low-risk, and robust marking method without causing the gene functions to deteriorate.

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mentioning

confidence: 99%

“Mark the Gene”: a Method for Nondestructive Introduction of Marker Sequences Inside the Gene Frame of Transgenes

Morono

Kitagawa

Kimura

et al. 2007

Appl Environ Microbiol

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Detection and identification of multiple genetically modified events using DNA insert fingerprinting

et al. 2009

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Current screening and event-specific polymerase chain reaction (PCR) assays for the detection and identification of genetically modified organisms (GMOs) in samples of unknown composition or for the detection of non-regulated GMOs have limitations, and alternative approaches are required. A transgenic DNA fingerprinting methodology using restriction enzyme digestion, adaptor ligation, and nested PCR was developed where individual GMOs are distinguished by the characteristic fingerprint pattern of the fragments generated. The inter-laboratory reproducibility of the amplified fragment sizes using different capillary electrophoresis platforms was compared, and reproducible patterns were obtained with an average difference in fragment size of 2.4 bp. DNA insert fingerprints for 12 different maize events, including two maize hybrids and one soy event, were generated that reflected the composition of the transgenic DNA constructs. Once produced, the fingerprint profiles were added to a database which can be readily exchanged and shared between laboratories. This approach should facilitate the process of GMO identification and characterization.

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Identity tags revisited

Cited by 2 publications

References 0 publications

“Mark the Gene”: a Method for Nondestructive Introduction of Marker Sequences Inside the Gene Frame of Transgenes

“Mark the Gene”: a Method for Nondestructive Introduction of Marker Sequences Inside the Gene Frame of Transgenes

Detection and identification of multiple genetically modified events using DNA insert fingerprinting

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