[Arabidopsis] Clone Donation Request

Associomics via arab-gen%40net.bio.net (by associomics from stanford.edu)
Thu Feb 17 17:59:26 EST 2011

> Clone Donation Request
> As part of our membrane/signaling protein interactome project called =
Associomics, we have completed the wet lab analysis of pairwise =
interactions (~6.4 million pairs tested).  We will in the near future =
release the data from this test (50% of the data are currently available =
through our website www.associomics.org). We plan on expanding the =
analysis to obtain a more complete interactome of membrane =
proteins/signaling proteins from Arabidopsis using the split-ubiquitin =
system. To do this, we need additional ORFs.
> The set of genes we intend to look at comprises most integral membrane =
proteins (>4,300, 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=92s quite an undertaking to clone all these =
genes and thus far we have >3500 successfully cloned genes, most of =
which have already been screened.  Some of these genes were donations =
from many of you and we are always grateful for such assistance.  We are =
now preparing for a continuation of the cloning stage and have a list of =
3298 genes that were not successfully obtained in the first rounds of =
cloning and a further 1755 genes that we would like to have but have not =
attempted cloning yet.  A file containing these gene lists can be found =
here:http://associomics.stanford.edu/request_donation.php.  Proteins =
localizing at the outer membrane of the chloroplast and having a =
cytosolic C terminus could also be included. Therefore, if you are =
willing to add any gene that fits in either category (integral membrane =
protein or signaling) that might be of interest for you and which we =
have not cloned yet, please contact Alexander Jones =
(associomics from stanford.edu) with questions or send clones directly to=20

Maria Sardi
Frommer Laboratory
Carnegie Institution Plant Biology
260 Panama St.
Stanford, CA 94305-4101 =20
> Any appropriate clone donation is welcome. =20
> All we need is an ORF cloned into a Gateway Entry vector, with ATG but =
without a 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 C-terminal ubiquitin fusion is inside an =
organelle, compartment or outside the cell (i.e. non-cytosolic).  If =
both termini of a protein are non-cytosolic, then it is not suitable for =
this screening system.
> For more information, please see the publication describing the =
results from a smaller pilot screen described here  =
ract). =20
> We will move the clones via gateway into the suitable destination =
vectors, carry out the screening and analysis and make the data from the =
subscreens available as fast as we can through the website =
www.associomics.org. =20
> Also, we would like to make the resulting split ubiquitin clones =
available through ABRC, so wherever possible, it would be great to get =
them without strings attached. =20

> NOTE: The clones are not yet available for distribution. We are =
planning to make them available through ABRC in the very near future.
Alexander Jones, Ph.D.
Frommer Laboratory
Carnegie Institution for Science
260 Panama St.=20
Stanford, CA 94305-4101

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