Cloning receptor genes using the ligand?

bbraun at ucla.edu bbraun at ucla.edu
Thu Nov 23 15:04:09 EST 1995


In <4921sh$fla at is.bbsrc.ac.uk>, michael.baron at bbsrc.ac.uk (Michael D. Baron) writes:
>In article <49079m$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

OK, I don't know from nothing, but another neat example might be the
cloning of the Flt3/Flk2 ligand.  The referece is <frantic database search>
Cell 75(6):1157.

Benjamin




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