MPE-Fe footprinting

Geoff Kornfeld geoffk at usage.csd.unsw.oz.au
Fri Jul 13 03:28:41 EST 1990



We are presently using Cu/phenanthroline to footprint the promoter of the
yeast LPD1 gene but have come across a region which does not give a good
ladder in the absence of protein. As luck would have it, this region 
is a transcriptional activator binding site (we know that from other work).

We would like to use the MPE-Fe footprinting system of Hertzberg & Dervan
to gte around this (and to give us a more precise picture of what is
happening at some other sites) but there are a few problems:


	1) Is there a commercial distributer of MPE-Fe? I have been
	   unable to find any here.

	2) Have those out there who have used this method (& I hope
	   that there are some) had to synthesise your own (if there
	   is no commercial source) or were you able to get it from
	   the authors?

	3) I am willing to synthesise it if necessary, but the Australian
	   branch of May & Baker does not have a listing for 
	   p-carboxymethidium chloride (5-methyl-6-(4-carboxylphenyl)-
	   3,8-diaminophenanthridinium chloride). [ the papers cite
	   them as the source ]. Is ther an alternative source?

I would appreciate it if anyone who has used this technique could help.



							Geoff Kornfeld
							School of Biochemistry
							UNSW



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