STOP ADVERTISING !!!
UZS13B at ibm.rhrz.uni-bonn.de
Mon Sep 19 11:19:57 EST 1994
Ram Balaraman thinks it's funny to pick up e-mail addresses of biologists
from this newsboard and answer their questions with disgusting comercials.
I asked that guy what he intents besides making money, but anything but
putting an order seems not to be worth an answer from him.
I think this abuse of the internet is worth a little bit of punishment,
but I don't know how to tell my IBM to resend that trash every 20 seconds
(certain lawyers know that kind of "mails").
Has anyone a better idea how to react or can do the job for me on his
Stefan Kahlert, Medizinische Poliklinik der Universitaet Bonn
| Stefan Kahlert |
| Phone D49-228-465731 |
| UZS13B at IBM.RHRZ.Uni-Bonn.de |
You want to become the hangman?
Fine, here is the rope:
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Date: Tue, 13 Sep 1994 07:45:40 +0059 (EDT)
From: Ram Balaraman <synapsys at world.std.com>
Subject: Multi-Functional CAT & Luciferase Reporter Gene Vectors
To: j.grimbergen at pg.tno.nl, schupp at nauvax.ucc.nau.edu,
umm500 at ibm.rhrz.uni-bonn.de
Message-Id: <Pine.3.89.9409130711.B58-0100000 at world.std.com>
Content-Type: TEXT/PLAIN; charset=US-ASCII
Near Zero-Background Multifunctional Reporter Gene Vectors For Rapid
Analysis of Eukaryotic Promoters & Enhancers.
The vectors , SV40-Syncat, Syncat I, Syncat II, SV40-pFlash,
pFlash I and pFlash II, are as their names suggest CAT & Luciferase
(Luc) reporter gene vectors. What distinguishes them from other vectors
in this category is that these vectors produce near-zero background.
This feature is due to the fact, that they use a modified SV40-t-intron
as the donor for the poly-adenylation signal. The wild type
SV40-t-intron is the generic donor for this function and is widely used
in a variety of reporter gene vectors, eg. pCAT- basic, pCAT-promoter,
pGL-basic and pGL-promoter (Promega). However one little known fact
about the wild-type-t-intron is that it contains cryptic enhancer
sequences that produce significant background activity, thereby raising
the threshold of sensitivity.
Until now this feature was not a serious limitation since scientists'
were analysing strong promoters/enhancers that produced high signal to
noise ratios. However the focus in the past couple of years has shifted to
(a) the analysis of weak promoters,
(b) promoters that have very low basal transcription rates but are
specifically induced to high levels by cytokines,
------- OR -------
(c) promoters that function only in cells that are very poorly transfectable.
Analysis of such regulatory elements requires systems of high sensitivity
as well as specificity.
The Syncat & pFlash series of vectors provide precisely these capabilities,
since they contain a t-intron polyA signal that has been modified to be
devoid of cryptic enhancer activity.
The 2nd. feature of these vectors is the choice of the heterologous promoter
in Syncat II and pFlash II. These vectors use the well characterized HSV-tk
TATA box containing minimal promoter situated immediately upstream of
the reporter gene. The HSV-tk is a widely used basal promoter that has
been documented to produce no spurious interactions with enhancers of interest
By contrast vectors like pCAT-promoter or pGL-promoter use the
SV40 promoter. The SV40 promoter contains an atypical TATA-box and more
seriously has been documented to spuriously repress certain cytokine
inducible enhancers (Benech, et.al.. J.Exp.Med. 1992, Vol. 176:
1115-1123). This latter anomalous property of the SV40 promoter is a
serious liability since it is impossible to be 100% confident that the
cis-elements under analysis will not be spuriously repressed by the SV40
heterologous promoter used in the pCAT/pGL vectors.
Other features of these vectors include a very versatile multiple cloning
site upstream and downstream of the reporter gene cassette, flanked by
T3 & T7 promoter sequences for direct sequencing and creation of
unidirectionally deleted mutant libraries as well as f1 origin of
replication for ssDNA recovery and site-directed mutagenesis.
The combination of these features enables you to perform all your DNA
manipulations from--eg., cloning of a 5 kb genomic 5U flanking region
upstream of the reporter gene ---to--- creation of nested deletion mutant
libraries followed by sequence verification ---to--- ssDNA-site-directed
mutagenesis and transfection of each construct --- ALL IN THE SAME
REPORTER GENE VECTOR YOU STARTED WITH. The time savings alone can be upto
IN TODAYS FAST PACED WORLD---THIS COULD WELL BE THE DIFFERENCE BETWEEN BEING
THE FRONT RUNNER & the runner up.
These vectors are available from SynapSys Corp. To get more info
about these vectors send E-mail to :-
synapsys at world.std.com
PLEASE INCLUDE :- Your COMPLETE POSTAL MAILING ADDRESS & preferably your
phone & fax # to enable us to mail/fax you any information or otherwise
Marketing & Sales
SynapSys Corp. -----We Bridge the Gap-----
7, Bradford Rd., Burlington, MA 01813
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