What is Kd? Also, in reply to earlier posts: it is sadly common in crystallizing large protein-DNA complexes to go through a couple dozen different duplexes and several dismally-diffracting crystal forms before finding a good one.
Phoebe >From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> (on behalf of umar farook ><umarfaroo...@gmail.com>) >Subject: [ccp4bb] Protein-DNA complex crystallization >To: CCP4BB@JISCMAIL.AC.UK > > Dear All, > I have been trying to crystallize protein DNA > complex, but all the time i end up with DNA > crystals. Even i changed the length of DNA many > times but still no complex, DNA only crystallizes! > Does anybody has idea, why do DNA crystallize by > itself ? My protein behaves very nicely, Dynamic > Light Scattering always shows nice values implies > homogenous but once i tried to ran acidic native > page but it shows little bit aggregated. The protein > is highly hydrophilic and soluble, and has only > three cysteines, is it there any possibility of > aggregation due to cysteine, when overexpressed in > E.coli ? and one more thing i mixed protein and DNA > together and ran agarose gel to see any gel shift, > indeed there is a binding, but when i take the same > thing to set drops, only DNA crystals. Kindly > suggest me, what could be done. > Thanks & Regards, > Umar Farook.S
