Translation initiation in drosophila

[Nico Stuurman]

Dear all,

I am planning to express a couple of point mutants of MFP (my favorite 
protein) in Drosophila using P-element mediated transformation.  I have 
expressed most of these mutants already in E. coli using the pET system.  
For cloning purposes it would be extremely useful to employ an NdeI (CATATG) 
site at the initiating methionine codon.  However, I don't know how 
detremental this will be for translation efficiency of the protein in 
Drosophila.  I checked a couple of Drosophila mRNAs in Genbank and none of 
them contains such a sequence; all have an A (or rarely a G, but never a C) 
at the -3 position.  Does anyone have experience with the effect on 
expression levels of altering the bases preceding the initiating methione 
codon?

Thanks for your help


Nico Stuurman

Dept. Pharmacological Sciences
SUNY at Stony Brook
Stony Brook, NY 11794-8651
T: 516 444 3079
F: 516 444 3218
Email: Nico at pharm.som.sunysb.edu