Clone Donation Request

As part of an Arabidopsis 2010 project, we plan to determine the potential interactome of membrane proteins from Arabidopsis using the split-ubiquitin system.

The set of genes we intend to look at comprises essentially all integral membrane proteins (>5,000, except chloroplast and mitochondrial proteins since we were afraid that they may be targeted to mitochondria in yeast and thus not be accessible for the interaction trap) and a large number (>1,000) of proteins predicted to be involved in signaling or protein modification or which might interact with the set of membrane proteins.

As you can imagine, it’s quite an undertaking to clone all these genes (current list of genes cloned), so are willing to add any gene that fits in either category that might be of interest for you and which we have not cloned yet. Proteins localizing at the outer membrane of the chloroplast and having a cytosolic C terminus could also be included.

Any clone donation would therefore be welcome.

All we need is an ORF cloned into a Gateway vector, with ATG but without stop codon at the end of the coding sequence. Moreover, any information about the topology of your proteins would be valuable, since the system will not work if the Cub-fusion is inside an organelle, compartment or outside the cell.

We will move the clones via gateway into the suitable destination vectors, carry out the analysis using robotic and make the data from the subscreens available as fast as we can. We hope to have the first set of data for 1000-2000 proteins in a binary matrix online before summer 2008.

NOTE: The clones are not yet available for distribution. We are planning to make them available through ABRC in the near future.