2012 Promega catalogue

Life
Science
Catalog
2012
Worldwide Contact List
Section
Contents
Table of
Contents
Cell Signaling
300
pAdVAntage Vector
Product Size Cat.# Price ($)
pAdVAntage Vector 20 µg E1711 167.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: Co-transfection of mammalian cells with the pAdVAntage
Vector enhances transient protein expression in a variety of cell types by
increasing translation initiation.
Transfection of mammalian cells with an expression vector often results in
suboptimal expression of the protein of interest. Double-stranded RNA (dsRNA)
generated during transfection is thought to activate the dsRNA-activated
inhibitor (DAI), one of several enzymes involved in the host cell’s antiviral
defense system. DAI phosphorylates the translation initiation factor eIF-2,
halting translation and therefore protein production.
However, DAI translation inhibition can be overcome with the adenoviral Virus
Associated I RNA (VAI RNA) produced by RNA polymerase III following co-transfection
with the pAdVAntage Vector. The VAI RNA binds to DAI, preventing its
activation, thereby allowing translation and protein expression.
Features:
• Increased Expression: Co-transfection of pAdVAntage Vector with
luciferase constructs showed at least a tenfold increase in luciferase
expression in 293 and HeLa cell lines over transfections performed with
the construct DNA alone.
• Flexible: Can be used in a variety of cell lines.
Storage Conditions: Store at –20°C.
Protocol Part#
pAdVAntage Vector Technical Bulletin TB207
pSI Mammalian Expression Vector
Product Size Cat.# Price ($)
pSI Mammalian Expression Vector
For Research Use Only. Not for Use in Diagnostic Procedures.
20 µg E1721 423.00
Description: The pSI Mammalian Expression Vector promotes constitutive
expression of cloned DNA inserts in mammalian cells. The major difference
between the pCI and pSI Mammalian Expression Vectors is the enhancer/
promoter region controlling the expression of the inserted gene. The pSI
Expression Vector contains the simian virus 40 (SV40) enhancer and early promoter
region. This vector can be used for both transient and stable expression
of genes. For stable expression, the pSI Vector must be co-transfected with an
expression vector containing a selectable gene for mammalian cells.
Features:
• Strong, Constitutive Expression: The pSI Vector’s SV40 enhancer/
promoter region allows strong, constitutive expression in most cell lines.
The vector is maintained as an episome in cells expressing the SV40 large
T antigen, leading to even higher levels of expression. A β-globin/IgG
chimeric intron located downstream from the enhancer/promoter region
can further increase expression.
• Increased Steady-State mRNA Levels: The late SV40 polyadenylation
signal increases the steady-state level of RNA approximately fivefold more
than the early SV40 polyadenylation signal.
• Convenient: Multiple cloning sites exist for easy insertion of cDNA.
• Versatile: Synthesize transcripts in vitro using the T7 RNA polymerase
promoter or generate single-stranded DNA in E. coli using the f1 origin of
replication.
Storage Conditions: Store at –20°C.
Protocol Part#
pCI and pSI Mammalian Expression Vectors Technical Bulletin TB206
Amp r
ori
pSI
Vector
(3632bp)
Intron
T7
NheI
XhoI
EcoRI
MluI
XbaI
SalI
AccI
SmaI
BstZI
NotI
➞
For complete and up-to-date product information visit: www.promega.com/catalog
f1 ori
BglII
SV40 Enhancer/
Early Promoter
SV40 Late
poly(A)
Bst98I
PstI
Bst98I
BamHI
678
684
689
695
707
713
714
720
724
724
0684VA06_4B