title: Development of a new pCAMBIA binary vector using Gateway® technology.
abstract: pCAMBIA vectors have become popular for their easy handling, stability and the existence of a range of selection and reporter genes. However, these vectors have yet to integrate the Gateway® cloning system, which has enabled site-specific recombination without the need for restriction enzymes and ligases. This paper sets out to convert the pCambia2300 binary vector into a destination vector with the Gateway® cassette driven by the CaMV35S promoter. The destination vector, pCamway35S, was then evaluated using the uidA reporter gene. Transient and stable transformation experiments were successfully assayed, either by particle bombardment or by Agrobacterium tumefaciens in Allium cepa and Hevea embryogenic calli. After counting the transformation units, the statistical analysis performed on the data showed that the pCamway 35S::uidA vector was as efficient as pCambia2301, a pCAMBIA2300 containing the uidA reporter gene under the CaMV 35S promoter.
authors.affiliation: CIRAD, UMR AGAP, F-34398 Montpellier, France. Electronic address: julie.leclercq@cirad.fr.
authors.forename: Julie
authors.initials: J
authors.lastname: Leclercq
authors.affiliation: CIRAD, UMR AGAP, F-34398 Montpellier, France. Electronic address: szabolcstoth70@yahoo.fr.
authors.forename: Toth
authors.initials: T
authors.lastname: Szabolcs
authors.affiliation: CIRAD, UMR AGAP, F-34398 Montpellier, France. Electronic address: florence.martin@cirad.fr.
authors.forename: Florence
authors.initials: F
authors.lastname: Martin
authors.affiliation: CIRAD, UMR AGAP, F-34398 Montpellier, France. Electronic address: pascal.montoro@cirad.fr.
authors.forename: Pascal
authors.initials: P
authors.lastname: Montoro
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D015870:Gene Expression; D023061:Gene Order; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D015203:Reproducibility of Results; D014169:Transformation, Bacterial
keywords: Genetic transformation; Green fluorescent protein