A new set of small, extrachromosomal expression vectors for Dictyostelium discoideum.
A new set of extrachromosomal Dictyostelium expression vectors is presented that can be modified according to the experimental needs with minimal cloning efforts. To achieve this, the vector consists of four functional regions that are separated by unique restriction sites, (1) an Escherichia coli replication region, and regions for (2) replication, (3) selection and (4) protein expression in Dictyostelium. Each region was trimmed down to its smallest possible size. A basic expression vector can be constructed from these modules with a size of only 6.8 kb. By exchanging modules, a large number of vectors with different properties can be constructed. The resulting set of vectors allows most basic expression needs, such as immuno blotting, protein purification, visualization of protein localization and identification of protein-protein interactions. In addition, two genes can be simultaneously expressed on one vector, which yields far more synchronous levels of expression than when expressing two genes on separate plasmids.
Veltman
Douwe M
DM
Cell Biochemistry, Department of Biology, University of Groningen, Haren, The Netherlands. d.veltman@beatson.gla.ac.uk
Akar
Gunkut
G
Bosgraaf
Leonard
L
Van Haastert
Peter J M
PJ
D000818:Animals; D003001:Cloning, Molecular; D004023:Dictyostelium / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology*; D010957:Plasmids / Q000235:genetics / Q000502:physiology*