Golden-Gate compatible Magnaporthe oryzae Agrobacterium transformation vectors

The Golden Gate cloning system uses standardised parts to facilitate the assembly of multiple transcriptional units, to ensure that future work with these genes can be carried out with ease (Patron et al., 2015 New Phytologist, v. 208, p. 13-19).<br><div><br></div><div>We have developed the Golden Gate compatible vector pBHt2G-RFP (Addgene #107162) from the pCAMBIA-derived (Mullins et al., 2001) pBHt2G vector (Khang et al, 2010). The vector was domesticated through removal of BsaI cloning sites. An RFP-marker was inserted, which is expressed in E. coli, allowing for red-white selection of transformants. The marker is lost during the Golden Gate reaction, as it is replaced by the inserted transcriptional units.</div><div><br></div><div>Vector, sequence information and plasmid maps are available from Addgene https://www.addgene.org/107162//</div

Golden-Gate compatible Magnaporthe oryzae protoplast transformation vectors

<div>The Golden Gate cloning system uses standardised parts to facilitate the assembly of multiple transcriptional units, to ensure that future work with these genes can be carried out with ease (Patron et al., 2015 New Phytologist, v. 208, p. 13-19).</div><div><br></div><div>Three fungal transformation vectors have been adapted from the pCB1532 vector series (Sweigard et al., 1997. Fungal Genetics Newsletter 44: 52-53). Vector pCB1532B-RFP Addgene #101854 encodes bialaphos/basta/L-phosphinothricin resistance, pCB1532H-RFP #101855 hygromycin resistance and pCB1532S-RFP #101856 sulfonylurea/chlorimuron ethyl resistance. </div><div><br></div><div>Vectors were domesticated through removal of BsaI cloning sites. An RFP-marker was inserted. The RFP is expressed in E. coli, allowing for red-white selection of transformants. The marker is lost during the Golden Gate reaction, as it is replaced by the inserted transcriptional units. Vectors, sequence information and plasmid maps are available from Addgene https://www.addgene.org/plasmids/articles/28191792/ </div><div><br></div

A Golden Gate Modular Cloning Toolbox for Plants

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 <i>Nicotiana benthamiana</i> is discussed