From owner-biotechniques@net.bio.net Wed Mar 18 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Lolis, Elias" Newsgroups: bionet.journals.letters.biotechniques Subject: mutagenesis strategy Date: 19 Mar 1998 13:09:24 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 18 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6es1i4$uu@net.bio.net> NNTP-Posting-Host: net.bio.net 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 .