Retics and P'ation

Steve Rodems smrodems at
Fri Apr 22 00:55:12 EST 1994

I have a suspected eukaryotic DNA binding protein that may require
phosphorylation for activity.  Rabbit reticulocyte lysates seem to
correctly phosphorylate the protein but I'd like to get my hands on more
protein than retics provide.  Can I use retics not for translation but just
for post-translational modification, ie, can I add protein to retic lysates
and get phosphorylation or are translation and phosphoylation in some way
coupled so that only nascent peptides are phosphorylated?  This may be
basic biochem but if I ever learned it I forgot it!  I could use
baculovirus expression but that takes a lot of time.  Any other suggestions
on how to get a lot of correctly phosphorylated protein in vitro?

Steve "Some day I will get the hell out of Wisconsin" Rodems

"Then I am here for the Lee family renioun ... shur-wajo-shur"

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