Dear All,
First I would like to thanks everyone who took the time to respond. It never
fails to amaze me what an awesome forum the ccp4bb board is and how willing
people are to share their expertise!
Below is a summary of the replies I received
1. Do people routinely try different
I was hoping people could give some tips on the best way to go about
crystallizing a protein-DNA complex.
I have a large amount of experience in protein crystallisation but have
never tried co-crystallisation with DNA until I started this project.
If you want to reply to me personally I will then
Hi Clare,
Two papers that might be worth checking out that address some of your
questions -
1. Tan et al, Crystallization of the Yeast MATa2/MCM1/DNA Ternary Complex:
General Methods and Principles for Protein/DNA Cocrystallization J. Mol.
Biol. (2000) 297, 947±959
2. Cannone et al,
1. Do people routinely try different lengths of DNA?
Yes, its usually the most important variable because the ends
like to stack against each other to form a pseudo-continuous
helix.
2. Do you start with blunt or sticky ends?
Both. Plan on a dozen or so different duplexes to start
Dear Clare et al:
The absolute classic paper in the field is this:
Schultz, S. C., Shields, G. C. Steitz, T. A. (1990).
Crystallization of Escherichia coli catabolite gene
activator protein with its DNA binding site. J. Mol.
Biol. 213, 159–166.
I've learned more from this than from reading
