Greetings Steven,
I had a bunch of mutants and may not have used those lines in the paper. Here is the information you need:
1. They are standard UAST constructs and the changes are via PCR based mutagenesis. The plasmids were made by D. E Kim (the other first author on that paper ) and we did the injections at Model Systems Genomics, Duke University.
2. The RagC S45N is a dominant negative mutant. The amino acid change results in a RagC protein where no nucleotide can bind to this small G protein (it needs to be in the GTP bound form to be active and GDP bound form is inactive).
3. The RagC Q99L is a constitutively active mutant. The amino acid change results in a RagC protein which always stays in the GTP bound form.
regards,
Pankuri Goraksha-Hicks
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On May 12, 2011, at  4:36  AM, Steven Marygold wrote:
Dear Pankuri Goraksha-Hicks,
I am curating a paper for FlyBase by Resnik-Docampo and de Celis ("MAP4K3 is a component of the TORC1 signalling complex that modulates cell growth and viability in Drosophila melanogaster.", FBrf0212911) in which the authors use some fly lines donated by you.
These are:
UAS-RagCQ99L
UAS-RagCS45N
I've been in contact with Martin Resnik-Docampo and he told me these stocks are described in your NCB paper "Regulation of TORC1 by rag GTPases in nutrient response" (FBrf0206857).  However, I'm afraid I can't see them described/used in that paper.
Before I make new entries for these transgenes in FlyBase, I need to check their molecular composition.
I assume that they are standard UAST-based constructs.  I think you used the GH16429 EST and introduced the Q99L and S45N mutations by PCR-based mutagenesis.  Is that correct?
Can you also tell me what effect these mutations are predicted to have on the protein - dominant negative, dominant active?
Thanks very much for your time,
Steven Marygold.