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GreenGate - TypeIIS RE Cloning System for Plant Transgenesis

Written by Dr. Marcelo Lanz | May 14, 2021 8:13:00 AM

Lampropoulos A, Sutikovic Z, Wenzl C, Maegele I, Lohmann JU, Forner J (2013) GreenGate - A Novel, Versatile, and Efficient Cloning System for Plant Transgenesis. PLoS ONE 8(12): e83043. https://doi.org/10.1371/journal.pone.0083043

The Golden Gate cloning method is an easy and straight forward procedure. Here, Lampropoulos et al. describe a variety of it designed for plant transgenesis, calling it cleverly GreenGate. The whole procedure is possible due to type IIS restriction enzymes. These enzymes cut at a defined number of nucleotides after the recognition site. Our FastGene BsaI is such an enzyme.

The plant transformation vectors created for this system include:

  • Promoter
  • N-terminal tag
  • Coding sequence (CDS) (i.e. the gene of interest)
  • C-terminal tag
  • Plant terminator
  • Plant resistance cassette

 The major advantage of using GreenGate is a dramatic reduction of plasmid generation time when building a multi-construct transgenic plant.

Would you like to read the paper?

Just click on the link below.

GreenGate Paper

 

More information about the Bsa I Enzyme:

FastGene Restriction Enzyme Bsa I