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From: werner@aecom.YU.EDU (Craig Werner)
Newsgroups: sci.bio
Subject: Re: Cloning/Expression vector copy numbers
Message-ID: <1161@aecom.YU.EDU>
Date: Fri, 26-Jun-87 02:02:05 EDT
Article-I.D.: aecom.1161
Posted: Fri Jun 26 02:02:05 1987
Date-Received: Sun, 28-Jun-87 00:56:55 EDT
References: <1137@aecom.YU.EDU> <404@uhnix2.UUCP>
Organization: Albert Einstein Coll. of Med., NY
Lines: 21

In article <404@uhnix2.UUCP>, bchso@uhnix2.UUCP (Dan Davison) writes:
> 
> Have you considered and rejected chloramphenicol amplification?  I've heard
> it can get 1000s (!) per cell.  The plasmid must be descended from ColE1,
> though.
> 
> dr. dan davison|Los Alamos National Laboratory|Theoretical Biology|T-10,

	Only if it retains the _rom_ gene, which is what is repressed
and leads to amplification.  Many laboratory derivatives of pBR322,
such as all the pUC derivatives, have had this gene deleted. As a result,
they cannot be chloramphenicol-amplified.  However, their basal copy
number does increase from the 20 or so of pBR322 to about 150-250, which
makes amplification a bit gratuitous.


-- 
	   Craig "Baby Doc" Werner   (future MD/PhD, 3 years down, 4 to go)
	     werner@aecom.YU.EDU -- Albert Einstein College of Medicine
              (1935-14E Eastchester Rd., Bronx NY 10461, 212-931-2517)
          "Disinformation is one thing, but misinformation is unforgiveable."