Subject: 	Re: Helping FlyBase
Hi Steven,
..
For the RNAi, yes, a sense and anti-sense fragment of dPLD was cloned 
into pUAST and the expression induced with appropriate drivers (such as 
the one listed below). The fragment spanned exon2-exon3 according to 
Hilde Janssens's notes, but I didn't see an exact description of the primers or 
nucleotide locations.
best regards, Mike
Steven Marygold wrote:
> Dear Michael and Mary,
>
>
> I am currently curating two of your papers for the FlyBase records:
> - LaLonde et al., 2005, J. Cell Biol. 169(3): 471--479.  Regulation of
> phototransduction responsiveness...
> - Lalonde et al., 2006, BMC Dev. Biol. 6(60).  A role for Phospholipase
> D...
>
>
> I have a couple of queries relating to these papers that I need to
> resolve before I can complete my curation (and add the data to the
> FlyBase records), and I'm hoping you can help.
>
>
..
> 2. In Fig3D of the 2006 paper, you show embryos expressing Pld RNAi.  I
> can't find any molecular details for this in the Methods section, but I
> assume that you made UAS lines that express an inverted repeat construct
> of part of the Pld mRNA sequence using a GAL4 line (probably
> P{GAL4-nos.NGT}40) to induce expression.
> Are these assumptions correct?  Also, any extra details you have on the
> UAS-RNAi construct(s) (eg. the part of the Pld mRNA/cDNA that was
> targetted) would also be welcome.
>
>
> Thanks for your help with this.
>
> Steven Marygold (for FlyBase)
-- 
Michael A. Frohman, M.D., Ph.D.
Professor & Director, MSTP
Dept. Pharmacology &
  Center for Developmental Genetics
438 CMM
SUNY Stony Brook
Stony Brook, NY  11794-5140