title: New yeast/E. coli/Drosophila triple shuttle vectors for efficient generation of Drosophila P element transformation constructs.
abstract: We have generated a set of novel triple shuttle vectors that facilitate the construction of Drosophila-P-element transformations vectors. These YED-vectors allow the insertion of any kind of sequence at any chosen position due to the presence of a yeast casette which ensures replication and allows for homologous recombination in Saccharomyces cerevisiae. As a proof of principle we generated several reporter constructs and tested them in transgenic flies for expression and correct subcellular localization. YED-vectors can be used for many purposes including promoter analysis or the expression of tagged or truncated proteins. Thus, time-consuming conventional restriction site based multi-step cloning procedures can be circumvented by using the new YED-vectors. The new set of triple shuttle vectors will be highly beneficial for the rapid construction of complex Drosophila transformation plasmids.
authors.affiliation: University of Osnabrück, Faculty of Biology, Department of Zoology and Developmental Biology, Barbarastr. 11, D-49076 Osnabrück, Germany. paululat@biologie.uni-osnabrueck.de
authors.forename: Achim
authors.initials: A
authors.lastname: Paululat
authors.forename: Jürgen J
authors.initials: JJ
authors.lastname: Heinisch
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D017931:DNA Primers; D004330:Drosophila / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D017930:Genes, Reporter; D005822:Genetic Vectors; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D017931:DNA Primers