E. coli LacZ gene

news at news.net.uni-c.dk news at news.net.uni-c.dk
Mon Sep 13 08:40:00 EST 1999


No offence! My reply was ment to be debatable, not as an accusation. I use LacZ as a
reporter, but has done a lot of searching before i found the sequence that i choose
as the best (it propably isn't!). I am about to clone the gene in the
reportervector, but i have to remove a BamHI site in the process. This leads to a
sequence which is nor wildtype neither greatly modified.
My point is that this approach could yield a reporter of greater as well as lesser
functionallity, compared to the wildtype. In case of a stronger reporter this would
be the prefered choice. Eventhough a stronger reporter is found, there may be many
sligthly/medium modified constructions that are even better.
Another issue of great importance is the substrate of the beta-gal. A better assay
could be obtained by trying out new substrates. This approach might be the simplest
after all.

Troels

biosci-request at net.bio.net, "Dr. Duncan Clark" wrote:

> In article <7rb0k9$odb at net.bio.net>, news at news.net.uni-c.dk writes
> >Maybe the wildtype isn't the best reporter. Some kind of modified protein could
> >be better.
>
> Suggestions?
>
> >The proposed LacZ from Promega could be the answer, but it is not
> >necessarily good, just because they sell it!!!!
>
> I didn't suggest that it was good, just that it is available. I have no
> relation with Promega, I just noticed it in their catalogue. I'm sure
> others are available. One just has to search.
>
> Duncan
> --
> The problem with being on the cutting edge is that you occasionally get
> sliced from time to time....
>
> Duncan Clark
> DNAmp Ltd.
> Tel: +44(0)1252376288
> FAX: +44(0)8701640382
> http://www.dnamp.com
> http://www.genesys.demon.co.uk

--

***************************************************************
Troels Felding                                  (+45) 3327 5309
Carlsberg Research Laboratory       Fax:        (+45) 3327 4764
Department of Yeast Breeding        E-mail:     tfe at crc.dk
Gamle Carlsberg Vej 10
DK-2500 Copenhagen Valby
D E N M A R K
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