Microarray-based method for detection of unknown genetic modifications.
BMC Biotechnol
; 7: 91, 2007 Dec 18.
Article
en En
| MEDLINE
| ID: mdl-18088429
ABSTRACT
BACKGROUND:
Due to the increased use of genetic modifications in crop improvement, there is a need to develop effective methods for the detection of both known and unknown transgene constructs in plants. We have developed a strategy for detection and characterization of unknown genetic modifications and we present a proof of concept for this method using Arabidopsis thaliana and Oryza sativa (rice). The approach relies on direct hybridization of total genomic DNA to high density microarrays designed to have probes tiled throughout a set of reference sequences.RESULTS:
We show that by using arrays with 25 basepair probes covering both strands of a set of 235 vectors (2 million basepairs) we can detect transgene sequences in transformed lines of A. thaliana and rice without prior knowledge about the transformation vectors or the T-DNA constructs used to generate the studied plants.CONCLUSION:
The approach should allow the user to detect the presence of transgene sequences and get sufficient information for further characterization of unknown genetic constructs in plants. The only requirements are access to a small amount of pure transgene plant material, that the genetic construct in question is above a certain size (here >/= 140 basepairs) and that parts of the construct shows some degree of sequence similarity with published genetic elements.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Oryza
/
Análisis Mutacional de ADN
/
Sondas de ADN
/
Análisis de Secuencia de ADN
/
Arabidopsis
/
ADN de Plantas
/
Análisis de Secuencia por Matrices de Oligonucleótidos
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
BMC Biotechnol
Asunto de la revista:
BIOTECNOLOGIA
Año:
2007
Tipo del documento:
Article
País de afiliación:
Noruega