ZFIN ID: ZDB-PUB-170809-10
A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications
Fowler, D.K., Stewart, S., Seredick, S., Eisen, J.S., Stankunas, K., Washbourne, P.
Date: 2016
Source: PLoS One 11: e0159277 (Journal)
Registered Authors: Eisen, Judith S., Stankunas, Kryn, Stewart, Scott, Washbourne, Philip
Keywords: Zebrafish, Expressed sequence tags, Recombination reactions, Vector cloning, Lentivirus, Neurons, Membrane proteins, Cloning
MeSH Terms:
  • Animals
  • Chromatography, Affinity/methods*
  • Cloning, Molecular/methods*
  • DNA, Recombinant/genetics*
  • Embryo, Nonmammalian/cytology
  • Embryo, Nonmammalian/metabolism
  • Gene Expression*
  • Genetic Vectors*
  • Green Fluorescent Proteins/metabolism
  • HEK293 Cells
  • Hippocampus
  • Humans
  • RNA Interference*
  • Rats
  • Recombination, Genetic*
  • Zebrafish/growth & development
  • Zebrafish/metabolism
PubMed: 27500400 Full text @ PLoS One
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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.
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