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 | Herausgeber: | ELSEVIER SCIENCE BV | Journal: | GENE | Volumen: | 511 | Ausgabe: | 2 | Startseite: | 300 | Seitenende: | 305 | Zusammenfassung: | 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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geprüft am 18.05.2024