A versatile set of tagged expression vectors to monitor protein localisation and function in Dictyostelium.
We describe here a series of vectors for ectopic expression of tagged proteins in Dictyostelium discoideum. These vectors allow the addition of N- or C-terminal tags (GFP, mRFP, 3xFLAG, 3xHA, 6xMYC or TAP) with an optimised polylinker sequence and no additional amino acid residues at the N- or C-terminus of the protein. The expression cassettes were introduced into vectors containing Blasticidin or Geneticin resistance markers and into integrating as well as extrachromosomal plasmids. The vectors are designed as high and low copy versions and thus allow for a limited expression level control. They are also convenient with regard to complementation, co- and super-transformation. Finally the vectors share standardised cloning sites, so that a gene of interest can be easily transferred between vectors as experimental requirements evolve. These vectors were used to study the localisation of several putative RNA processing proteins including EriA and DicerB.
Dubin
Manu
M
Department of Genetics, University of Kassel, Heinrich-Plett-Str. 40, 34132 Kassel, Germany. manudubin@yahoo.com
Nellen
Wolfgang
W
D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004023:Dictyostelium / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D010957:Plasmids; D018741:Replication Origin
D005656:Fungal Proteins; D049452:Green Fluorescent Proteins