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Non-antibiotic, efficient selection for alfalfa genetic engineering.
Rosellini, Daniele; Capomaccio, Stefano; Ferradini, Nicoletta; Savo Sardaro, Maria Luisa; Nicolia, Alessandro; Veronesi, Fabio.
Afiliación
  • Rosellini D; Dipartimento di Biologia Vegetale Biotecnologie Agroambientali e Zootecniche, Università degli Studi di Perugia, Borgo XX giugno 74, 06121 Perugia, Italy. roselli@unipg.it
Plant Cell Rep ; 26(7): 1035-44, 2007 Jul.
Article en En | MEDLINE | ID: mdl-17333020
A selectable marker gene (SMG), usually conferring resistance to an antibiotic or herbicide, is generally introduced into the plant cells with the gene(s) for the trait of interest to allow only the cells that have integrated and express the foreign sequences to regenerate into a plant. The availability of several SMGs for each plant species is useful for both basic and applied research to combine several genes of interest in the same plant. A selection system based on gabaculine (3-amino-2,3-dihydrobenzoic acid) as the selective substance and the bacterial hemL gene [encoding a mutant for of the enzyme glutamate 1-semialdehyde aminotransferase (GSA-AT)] as the SMG was previously used for genetic transformation of tobacco. The hemL gene is a good candidate for a safe SMG, because GSA-AT is present in all plants and is likely involved in one metabolic step only, so that unintended effects of its overexpression in plants are not probable. In this work, we have compared this new selection system with the conventional, kanamycin-based system for alfalfa Agrobacterium-mediated transformation. The hemL and NptII genes were placed together into a T-DNA under the control of identical promoters and terminators. We show that the gabaculine-based system is more efficient than the conventional, kanamycin-based system. The inheritance of hemL was Mendelian, and no obvious phenotypic effect of its expression was observed.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Plantas / Selección Genética / Ingeniería Genética / Medicago sativa Idioma: En Revista: Plant Cell Rep Asunto de la revista: BOTANICA Año: 2007 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Alemania
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Plantas / Selección Genética / Ingeniería Genética / Medicago sativa Idioma: En Revista: Plant Cell Rep Asunto de la revista: BOTANICA Año: 2007 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Alemania