In article <49079m$rfb at news.cais.com>, rover at ns1.livnet.com (Rover) says:
>>> We have heard in the wind that there is a method somewhere for
>cloning receptor genes using the ligand or ligand information. If
>anyone has heard of this or knows of a reference please let me know.
There are a variety of techniques for expression cloning receptors,
usually based on using a sensitive method for detecting the ligand, such
as radioiodinating it. Sorry, I don't have the references to hand. The
paper that I remember being referred back to by a lot of others was in
Cell or EMBO J in 1989. The basic idea is that you transfect cells with
pools from your cDNA library in pCDM8 or similar, incubate with the
labelled ligand, then dip the cells in photoemulsion and expose for a
week or so.
If you have an antibody to the ligand that does not interfere with
binding you could try that using the CELICS system published in Cell (I
think) by Almond's group in the last year or so.
The yeast system is a no-no, I fear, since it will not work with surface
membrane proteins, or ligands that are not proteins.
If you can't find what you want soon, come back to me and I'll send you
the references I have (I have just been looking into this, as I also want
to clone a receptor, and have only the ligand).
michael.baron at bbsrc.ac.uk