[Protein-analysis] Is there a way to enhance yield of soluble DNA-binding protein in limiting DNA-protein interactions ?

Nicolas BAZEILLE via proteins%40net.bio.net (by nicolas.bazeille from curie.u-psud.fr)
Mon Feb 4 10:39:41 EST 2008


hi,


I try to purify a protein from yeast to assay some structural studies. 
unfortunatly, the yield is really poor and I want to find some ways to 
increase it. First, I change the medium and the temperature and the time of 
induction but I think that now it is optimal.
Because my protein is a DNA binding protein, I think that I lost some 
proteins fixed on genomic DNA that are found in the pellet during the step 
of clarification just after breaking cells. Does it may be right ? Is there 
a way to prevent this interactions (high salt concentration, additives ??)
I ask also, to know if there is a special manipulation to get a clear 
pellet during the step of cells collecting because my pellet is rapidly 
resolublize and the end of the run in spite of an optimal speed, resulting 
in the loss of some cells. Is there specials additives to get clear, 
compact and well packed pellet of cells, that I can put directly on the 
medium (YNB -URA galactose) at the end of induction.


Nicolas pHD student Institut Curie Paris france




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