RRC ID 30942
Author Veltman DM, Keizer-Gunnink I, Haastert PJ.
Title An extrachromosomal, inducible expression system for Dictyostelium discoideum.
Journal Plasmid
Abstract Inducible expression systems are essential for the expression of toxic proteins and are very convenient for proteins that induce strong side effects such as retardation of growth or development. Currently available systems for use in Dictyostelium either do not have a very tight control over expression levels or use a combination of an integrating and an extrachromosomal vector. We designed a new vector in which all components of the available 2-plasmid tetracycline-inducible system were combined onto a single extrachromosomal vector. Two types of inducible plasmids are presented, in which transcription is induced by adding or removing doxycycline, respectively. The location and orientation of the components was optimized in order to obtain a low background expression combined with high inducibility. The resulting vectors have a very low expression in the uninduced state (>1000-fold lower expression compared to that resulting from the act15 promoter), show a 10,000-fold induction of gene expression in a doxycycline concentration-dependent manner and are comparatively small (8.5 kb). With these new vectors, inducible gene expression is as easy as constitutive gene expression.
Volume 61(2)
Pages 119-25
Published 2009-3-1
DOI 10.1016/j.plasmid.2008.11.002
PII S0147-619X(08)00104-2
PMID 19046986
MeSH Animals Dictyostelium / genetics* Doxycycline / pharmacology* Gene Expression / drug effects* Gene Expression Regulation / drug effects Genetic Vectors* / drug effects Plasmids* Promoter Regions, Genetic / drug effects Protein Biosynthesis* / drug effects Tetracycline / pharmacology
IF 2.805
Times Cited 18
Cellular slime molds G90055 G90056 G90057 G90058 G90059 G90060 G90061 G90062 G90063 G90064 G90065 G90066