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
