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A chemical-inducible Cre-LoxP system allows for elimination of selection marker genes in transgenic apricot

文献类型： 外文期刊

作者： Petri, Cesar 1 ; Lopez-Noguera, Sonia 1 ; Wang, Hong 1 ; Garcia-Almodovar, Carlos 1 ; Alburquerque, Nuria 1 ; Burgo 1 ;

作者机构： 1.CEBAS CSIC, Dept Mejora Vegetal, Grp Biotecnol Frutales, Murcia 30100, Spain

2.Gansu Acad Agr Sci, Inst Fruit & Floriculture Res, Lanzhou 730070, Peoples R China

关键词： Fruit trees;Genetic engineering;Marker-free;Prunus armeniaca;Site-specific recombination;beta-Estradiol

期刊名称：PLANT CELL TISSUE AND ORGAN CULTURE （ 影响因子：2.711； 五年影响因子：2.73 ）

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收录情况： SCI

摘要： Transgenic plant development relies on the introduction of marker genes along with the gene(s) of interest to select and/or identify transgenic regenerants. Due to public concerns and regulatory issues, it would be advantageous to eliminate these marker genes once they are no longer needed. The chemical-inducible Cre-LoxP system is especially suitable for clonally-propagated plants, such as fruit trees, as no sexual crosses or rounds of transformation are required for marker-gene elimination. In this study, four transgenic pX6-GFP apricot (Prunus armeniaca L.) (cv. Helena) lines, carrying the gfp reporter gene encoding for the green fluorescent protein, were obtained following Agrobacterium tumefaciens-mediated transformation of leaf explants. The DNA site-specific recombination was precise and tightly controlled by the inducer beta-estradiol. Expression of the gfp gene was only detected when 3 mu M beta-estradiol was added to the medium. When nodal explants were incubated on a meristem development medium supplemented with 3 mu M beta-estradiol, marker gene elimination was observed in buds of all four transgenic lines, at an average frequency of 11.3 %, based on GFP expression. Further molecular analyses of four GFP-positive shoots, a single shoot from each transgenic line, revealed that DNA recombination was complete in two of shoots, but incomplete in the other two shoots.

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