Transgenic resistance to rhizomania virus [in sugar beet]
2004
Nagl, N.
Coat protein gene of beet necrotic yellow vein virus, causing agent of rhizomania, was isolated from inoculated leaves of sugar beet, C. quinoa and T. expansa, by RT-PCR and imuno capture RT-PCR. Gene was isolated with and without its leader sequence and cloned in the plasmid vector pUC 18. After their identity has been confirmed, cloned in the plasmid vector pUC 18. After their identity has been confirmed, cloned fragments were re-cloned into two modified plant transformation vectors: pCAMBIA3301M, with gene for resistance to gluphosinate ammonium (bar gene) as selectable marker and pCAMBIA1304M, with the resistance to hygromycin. Multicloning site and start codon at the beginning of the gus gene have been removed and fragments containing coat protein gene were cloned on the place of gus, i.e. gus and mgfp genes. Three constructors were made with each vector: CPL, containing coat protein gene with leader sequence; CPS, with gene for coat protein and CPSas with coat protein gene in antisense orientation. All constructs were transfered in the A.tumefaciens strain LBA4404. In order to investigate ability of created constructs to get incorporated in plant genome, transformation of tobacco and N. excelsior were done, as well as some preliminary research in sugar beet transformation.
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