pZERO vectors: pros and cons?

vilimf01 at mcrcr6.med.nyu.edu vilimf01 at mcrcr6.med.nyu.edu
Sat Jan 25 11:10:38 EST 1997


In article <Pine.SOL.3.91.970116160924.28835A-100000 at hsph.harvard.edu>, Dan Gebow <dgebow at hsph.harvard.edu> writes:
> On 16 Jan 1997, Paul N Hengen wrote:
> 
>> Can anyone with experience with the ccdB containing vectors such
>> as pZERO give their view on the pros and cons of using it?
>> I've heard that it is just great, but are there any drawbacks?
> 
> I use the pZERO - neo vector quite alot. I've cloned everything from 81bp 
> to 2000+ w/ it. The only drawback is making the IPTG plates, they are 
> expensive and don't last long.
> 

You may want to try DH5 alpha.  Ive tested them and they have a leaky
enough lac repressor that they don't require any IPTG on the plates to
kill the non recombinants.  Also, Invitrogen has a strain that lacks
the lac repressor (INV alpha F' i believe) that they send with their
TA kits.  I don't know why they don't send this strain with the
p-zero, but that would also remove the need for IPTG in the plates. 
The vector performs quite well in my hands, although I have had less
luck with shorter inserts (<100bp).  The major drawbacks are that you
have to buy the vector from invitrogen, and not grow up, or alter the
vector.  I believe that you are also supposed to inform invitrogen
every time you send an insert containing plasmid to anyone else.  The
vector is non optimal in that it doesn't have opposed T7 and SP6
promoters, and it has 2 BstX I sites in the polylinker that are
useless because the ccdB gene has one as well.  With all the imposed
restrictions on its use, it is not my first choice for a vector.  I
played around with it for a while, till I used up the amount that they
sent me, but I didn't order it again.  On principle, I don't think
that you should have to buy a particular plasmid vector more than
once, and I don't need the extra paperwork.

					usual disclaimers	SVEN






> DG
> 
> 
> 
>> 
>> --
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