New yeast/E. coli/Drosophila triple shuttle vectors for efficient generation of Drosophila P element transformation constructs

Autor(en): Paululat, Achim 
Heinisch, Juergen J.
Stichwörter: Drosophila; E. coli; EXPRESSION; FRAGMENTS; GENETIC-TRANSFORMATION; Genetics & Heredity; HOMOLOGOUS RECOMBINATION; MELANOGASTER; P-element; PHI-C31; Triple shuttle vector; Yeast
Erscheinungsdatum: 2012
Journal: GENE
Volumen: 511
Ausgabe: 2
Startseite: 300
Seitenende: 305
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. (C) 2012 Elsevier B.V. All rights reserved,
ISSN: 03781119
DOI: 10.1016/j.gene.2012.09.058

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