mutagenesis strategy

Lolis, Elias LolisE at MASPO3.MAS.YALE.EDU
Tue Feb 3 14:50:52 EST 1998


   I would like to randomly mutate all 7 transmembrane regions of G 
protein-coupled receptors and then select for mutated receptors with 
functional properties. I was wondering whether anyone knows of any method 
that could target different regions of a cDNA for simultaneous random 
mutagenesis. One strategy I thought of requires a mutagenesis method that 
selectively targets either single-stranded or double-stranded DNA. For 
example, if there is a method that selectively targets single-stranded DNA, 
I could produce a single-stranded version of the cDNA, protect the 
non-transmembrane regions by synthesizing complementary DNA for these 
sequences, and applying the mutagenesis procedure to the remaining 
single-stranded DNA. If anyone knows of a method or article that answers 
this question please send me e-mail. Thanks.

Elias Lolis 



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