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From: diaz@aecom.YU.EDU (Dizzy Dan Diaz)
Newsgroups: sci.bio
Subject: Using the slow form of Bal 31 Nuclease
Message-ID: <1445@aecom.YU.EDU>
Date: Thu, 12-Nov-87 17:47:38 EST
Article-I.D.: aecom.1445
Posted: Thu Nov 12 17:47:38 1987
Date-Received: Sun, 15-Nov-87 18:49:42 EST
Organization: Graduate School of Hard Knocks
Lines: 22
Keywords: Deletion analysis, controlled digestions

I am interested in putting a gene of interest in front of an inducible
promoter.  The polylinker on the vector has lots of sites, but getting
my gene right in front of the ribosome-binding sequence will either
require using oligonucleotide engineering of my gene (yuck! the time,
the expense, what a pain!), or a controlled digestion of my gene's 
promoter and subsequent ligation to the vector (lots of screening, but
not as much work as messing with oligos).

Bal 31 nuclease as prepared is much too difficult to control for my
purposes.  As sold by most firms, Bal 31 is a mixture of two distinct
polypeptides, the so-called "fast" and "slow" forms.  IBI is the only
company I know of that will sell you either form, or a mixture of the
two.  It may be that this is the sort of enzyme I have been looking for,
but I am uncertain, since I know of no one who is experienced with this
"slow" form of Bal 31.

If anyone can be of assistance with this matter, I would appreciate some
helpful email.  Thanks.

-- 
      dn/dx      Dept Molecular Biology   diaz@aecom.yu.edu
     Dan Diaz      Albert Finkelstein College of Quackery