A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications

Recombination-based cloning is a quick and efficient way to generate expression vectors. Recent advancements have provided powerful recombinant DNA methods for molecular manipulations. Here, we describe a novel collection of three-fragment MultiSite Gateway cloning system-compatible vectors providin...

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Bibliographic Details
Published in:PloS one 2016-08, Vol.11 (8), p.e0159277
Main Authors: Fowler, Daniel K, Stewart, Scott, Seredick, Steve, Eisen, Judith S, Stankunas, Kryn, Washbourne, Philip
Format: Article
Language:English
Subjects:
Animals
Biological research
Biology and Life Sciences
Chromatography, Affinity - methods
Cloning
Cloning vectors
Cloning, Molecular - methods
Deoxyribonucleic acid
Dimerization
DNA
DNA, Recombinant - genetics
Embryo, Nonmammalian - cytology
Embryo, Nonmammalian - metabolism
Embryos
Enzymes
Expression vectors
Fluorescence
Fluoroscopic imaging
Gene Expression
Genetic engineering
Genetic research
Genetic Vectors
Green Fluorescent Proteins - metabolism
HEK293 Cells
Hippocampus
Humans
Laboratory animals
Medicine and Health Sciences
Molecular biology
Neurons
Neurosciences
Physical Sciences
Plasmids
Proteins
Purification
Rats
Recombinant DNA
Recombination
Recombination, Genetic
Research and Analysis Methods
Ribonucleic acid
RNA
RNA Interference
RNA-mediated interference
Technology application
Toolkits
Vertebrates
Zebrafish
Zebrafish - growth & development
Zebrafish - metabolism
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Description
Summary:Recombination-based cloning is a quick and efficient way to generate expression vectors. Recent advancements have provided powerful recombinant DNA methods for molecular manipulations. Here, we describe a novel collection of three-fragment MultiSite Gateway cloning system-compatible vectors providing expanded molecular tools for vertebrate research. The components of this toolkit encompass a broad range of uses such as fluorescent imaging, dual gene expression, RNA interference, tandem affinity purification, chemically-inducible dimerization and lentiviral production. We demonstrate examples highlighting the utility of this toolkit for producing multi-component vertebrate expression vectors with diverse primary research applications. The vectors presented here are compatible with other Gateway toolkits and collections, facilitating the rapid generation of a broad range of innovative DNA constructs for biological research.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0159277