A series of Dictyostelium expression vectors for recombination cloning

We describe a series of Dictyostelium expression vectors for recombination cloning using the Gateway technology. DNA fragments generated by high fidelity polymerase chain reaction are cloned by topoisomerase-mediated ligation, then recombined into any of several Dictyostelium expression vectors usin...

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Bibliographic Details
Published in:Plasmid 2006-11, Vol.56 (3), p.145-152
Main Authors: Thomason, Peter A., Brazill, Derrick T., Cox, Edward C.
Format: Article
Language:English
Subjects:
Animals
Bacteriophage lambda
Base Sequence
beta-Galactosidase
Cloning, Molecular - methods
Dictyostelium
Dictyostelium - genetics
DNA Primers
DNA Topoisomerases
Expression vectors
Gateway
Gene Expression
Genetic Vectors - genetics
Green Fluorescent Proteins
Molecular Sequence Data
Phage l
Polymerase Chain Reaction - methods
Promoter Regions, Genetic - genetics
Recombination cloning
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Description
Summary:We describe a series of Dictyostelium expression vectors for recombination cloning using the Gateway technology. DNA fragments generated by high fidelity polymerase chain reaction are cloned by topoisomerase-mediated ligation, then recombined into any of several Dictyostelium expression vectors using phage λ LR recombinase. No restriction enzymes are used in this procedure. Coding regions can be expressed from their own promoters, or from a strong actin 15 promoter as a native protein, or with an amino or carboxyl-terminal GFP fusion. Gene promoters of interest can be analyzed by controlled expression of GFP and β-galactosidase. These vectors allow for rapid and simple characterization of novel DNA, and are ideal for high-throughput studies.
ISSN:0147-619X
1095-9890
DOI:10.1016/j.plasmid.2006.04.001