Excision of a selectable marker gene in transgenic banana using a Cre/lox system controlled by an embryo specific promoter

Antibiotic and herbicide resistance genes have been used in transgene technology as powerful selection tools. Nonetheless, once transgenic events have been obtained their presence is no longer needed and can even be undesirable. In this work, we have developed a system to excise the selectable marke...

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Bibliographic Details
Published in:Plant molecular biology 2013-09, Vol.83 (1-2), p.143-152
Main Authors: Chong-Pérez, Borys, Reyes, Maritza, Rojas, Luis, Ocaña, Bárbara, Ramos, Adolfo, Kosky, Rafael G., Angenon, Geert
Format: Article
Language:English
Subjects:
Agricultural biotechnology
Bananas
Base Sequence
Biochemistry
Biomedical and Life Sciences
Gene expression
Genes, Plant
Genetic Markers
Genetic Vectors - genetics
Genetic Vectors - metabolism
Globulins - genetics
Globulins - metabolism
Integrases - genetics
Integrases - metabolism
Life Sciences
Molecular Sequence Data
Musa - genetics
Musa - metabolism
Oryza - genetics
Oryza sativa
Plant biology
Plant Pathology
Plant Proteins - genetics
Plant Proteins - metabolism
Plant Sciences
Plant Somatic Embryogenesis Techniques
Plants, Genetically Modified - genetics
Plants, Genetically Modified - metabolism
Promoter Regions, Genetic
Seeds - genetics
Seeds - metabolism
Transformation, Genetic
Transgenic plants
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Summary:Antibiotic and herbicide resistance genes have been used in transgene technology as powerful selection tools. Nonetheless, once transgenic events have been obtained their presence is no longer needed and can even be undesirable. In this work, we have developed a system to excise the selectable marker and the cre recombinase genes from transgenic banana cv. ‘Grande Naine’ ( Musa AAA). To achieve this, the embryo specific REG - 2 promoter was isolated from rice and its expression pattern in banana cell clumps, somatic embryos and regenerated plantlets was characterized by using a pREG2::uidA fusion construct. Subsequently, the REG - 2 promoter was placed upstream of the cre gene, conferring Cre functionality in somatic embryos and recombination of lox sites resulting in excision of the selectable marker and cre genes. PCR analysis revealed that 41.7 % of the analysed transgenic plants were completely marker free, results that were thereafter confirmed by Southern blot hybridization. These results demonstrate the feasibility of using developmentally controlled promoters to mediate marker excision in banana. This system does not require any extra handling compared to the conventional transformation procedure and might be useful in other species regenerating through somatic embryogenesis.
ISSN:0167-4412
1573-5028
DOI:10.1007/s11103-013-0058-8