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February 5, 2014ACS Synthetic Biology1,056 citations

A Golden Gate Modular Cloning Toolbox for Plants

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CECarola EnglerMYMark YoulesRGRamona Gruetzner

Key Points

  • Establish a standardized modular cloning toolkit based on Golden Gate assembly to streamline multigene construct generation for plant transformation.
  • Designed a standardized resource comprising cloning vectors and 96 modular genetic parts (promoters, tags, reporters, signals, markers, terminators).
  • Characterized and compared the functional performance of the modular parts in the model plant Nicotiana benthamiana.
  • Created a robust 96-part Golden Gate assembly library enabling precision modular construction of multigene plant expression vectors.
  • Demonstrated reliable and comparative in vivo expression profiles across diverse promoters, reporters, and regulatory signals in Nicotiana benthamiana.

Abstract

Plant Synthetic Biology requires robust and efficient methods for assembling multigene constructs. Golden Gate cloning provides a precision module-based cloning technique for facile assembly of multiple genes in one construct. We present here a versatile resource for plant biologists comprising a set of cloning vectors and 96 standardized parts to enable Golden Gate construction of multigene constructs for plant transformation. Parts include promoters, untranslated sequences, reporters, antigenic tags, localization signals, selectable markers, and terminators. The comparative performance of parts in the model plant Nicotiana benthamiana is discussed.

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Cite This Study

Engler et al. (2014) studied this question.

synapsesocial.com/papers/69dbcb2b7d378569a9835d0chttps://doi.org/10.1021/sb4001504
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