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. 2012 Dec 15;511(2):300-5.
doi: 10.1016/j.gene.2012.09.058. Epub 2012 Sep 28.

New yeast/E. coli/Drosophila Triple Shuttle Vectors for Efficient Generation of Drosophila P Element Transformation Constructs

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New yeast/E. coli/Drosophila Triple Shuttle Vectors for Efficient Generation of Drosophila P Element Transformation Constructs

Achim Paululat et al. Gene. .

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.

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