Path: utzoo!mnetor!uunet!munnari!murdu!sam
From: sam@murdu.OZ (Sam Ganesan)
Newsgroups: sci.bio
Subject: Re: Antisense-RNA-derived mutants
Message-ID: <1366@murdu.OZ>
Date: 8 May 88 08:23:56 GMT
References: <1784@aecom.YU.EDU>
Reply-To: sam@murdu.UUCP (Sam Ganesan)
Organization: Microbiology Dept, Melbourne Uni, Australia
Lines: 32

In article <1784@aecom.YU.EDU> diaz@aecom.YU.EDU (Dizzy Dan) writes:
>I am interested in generating mutant strains of E. coli in two genes
>I've been working with.  I don't want to mutagenize and hunt for mutants
>(yuck, what a mess!). 

  Hey Dan, Mutagenising and screening for mutants is not that much of a 
mess if you know what kind of a mutant you are looking for. Which are genes 
you are working with. I have been generating spontaneous and oligonucleotide
directed mutants in the E. coli gene tyrR and it has not been easy I must
admit but has not been that bad either.

>                      The method I use should be simple, effective, and
>specific to the genes in question.  What if I put an antisense code
>for the gene(s) on a plasmid and transform.  The antisense message would
>then prevent translation of the sense message, right?  I'd appreciate some
>advice, although, as always, Dan Davison may not respond.

I realise antisense RNA was touted as the means of regulation a few years
ago but it has quite a few problems. If you really need some references 
send  me a note and I probably can dig up some both for and against.
Just out of curiosity, which genes are you working with and what kind of 
mutants do you want?
						Sam Ganesan

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        Victoria 3052, Australia.
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