title: A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications.
abstract: 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.
authors: Institute of Molecular Biology, Department of Biology, University of Oregon, Eugene, Oregon, United States of America.|Institute of Neuroscience, Department of Biology, University of Oregon, Eugene, Oregon, United States of America.
authors: Daniel K
authors: DK
authors: Fowler
authors: Institute of Molecular Biology, Department of Biology, University of Oregon, Eugene, Oregon, United States of America.
authors: Scott
authors: S
authors: Stewart
authors: Institute of Neuroscience, Department of Biology, University of Oregon, Eugene, Oregon, United States of America.
authors: Steve
authors: S
authors: Seredick
authors: Institute of Neuroscience, Department of Biology, University of Oregon, Eugene, Oregon, United States of America.
authors: Judith S
authors: JS
authors: Eisen
authors: Institute of Molecular Biology, Department of Biology, University of Oregon, Eugene, Oregon, United States of America.
authors: Kryn
authors: K
authors: Stankunas
authors: Institute of Neuroscience, Department of Biology, University of Oregon, Eugene, Oregon, United States of America.
authors: Philip
authors: http://orcid.org/0000-0003-1851-1282
authors: P
authors: Washbourne
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D003001:Cloning, Molecular / Q000379:methods*; D004274:DNA, Recombinant / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006624:Hippocampus; D006801:Humans; D034622:RNA Interference; D051381:Rats; D011995:Recombination, Genetic; D015027:Zebrafish / Q000254:growth & development / Q000378:metabolism
chemical_list: D004274:DNA, Recombinant; D049452:Green Fluorescent Proteins