Cloning receptor genes using the ligand?

Michael D. Baron michael.baron at bbsrc.ac.uk
Thu Nov 23 09:53:05 EST 1995


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.
>
>Thanks.
>

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



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