2012 Promega catalogue

Life
Science
Catalog
2012
Worldwide Contact List
Section
Contents
Table of
Contents
Cell Signaling
220
pTargeT Mammalian Expression Vector
System
Product Size Cat.# Price ($)
pTargeT Mammalian Expression Vector
System 20 reactions A1410 569.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The pTargeT Mammalian Expression Vector System is a
convenient system for cloning PCR products and for expression of cloned PCR
products in mammalian cells. The vector is prepared by digestion with EcoRV
followed by addition of a 3´ terminal thymidine to each end. These single 3´-T
overhangs at the insertion site greatly improve the efficiency of ligation of
a PCR product into the plasmid in two ways. First, the overhangs prevent
recircularization of the vector; second, they provide a compatible overhang
for PCR products generated by certain thermostable polymerases. These
polymerases often add a single deoxyadenosine, in a template-independent
fashion, to the 3´-ends of amplified fragments. The pTargeT Vector also
contains a modified version of the coding sequence of the α-peptide of
β-galactosidase, which allows recombinants to be selected using blue/white
screening.The pTargeT Vector carries the human cytomegalovirus (CMV)
immediate-early enhancer/promoter region to promote constitutive expression
of cloned DNA inserts in mammalian cells. This vector also contains the neomycin
phosphotransferase gene, a selectable marker for mammalian cells. The
pTargeT Vector can be used for transient expression or for stable expression
by selecting transfected cells with the antibiotic G-418.
Features:
• Simple PCR Cloning: “T” overhangs permit direct ligation of PCR
products generated by thermostable enzymes such as Taq DNA
polymerase.
• Strong, Constitutive Expression: The CMV enhancer/promoter region
allows strong, constitutive expression in many cell types. In transgenic
mice, expression of the chloramphenicol acetyltransferase (CAT) gene
under the regulation of the CMV enhancer/promoter was observed in
24 of the 28 tissues examined. The vector is maintained as an episome
in cells expressing the SV40 large T antigen, leading to even higher levels
of expression.
• Blue/White Screening: Allows the easy identification of recombinant
clones. A single digest removes the insert DNA.
• Stable Transfectants: Select for stable transfectants using the neomycin
phosphotransferase gene.
Storage Conditions: Store competent cells at –70°C; store all other
components at –20°C or –70°C.
Protocol Part#
pTARGET Mammalian Expression Vector System Technical Manual TM044
Ampr
ori
Synthetic
poly(A)
Neo
BglII 5665
CMV
Enhancer/Promoter
pTARGET
Vector
(5670bp)
Intron
fl ori
SV40 Enhancer/
EarlyPromoter
SgfI 664
SV40 Late
poly (A)
I-PpoI
851
T
T
lacZ
T7
EcoRI
BamHI
NheI
XhoI
MluI
SmaI
KpnI
SalI
AccI
NotI
EcoRI
lacZ
➞
T overhangs
1250
1256
1264
1270
1276
1293
1301
1303
1304
1311
1318
1505VA07_6A
pTargeT Sequencing Primer
Product Size Cat.# Price ($)
pTargeT Sequencing Primer
For Research Use Only. Not for Use in Diagnostic Procedures.
2 µg Q4461 134.00
Description: The pTargeT Sequencing Primer is designed for sequencing
inserts cloned into the pTargeT Mammalian Expression Vector (Cat.# A1410).
The sequencing primer hybridizes to the region of the lacZ gene from
nucleotides 1367–1344 on the pTargeT Vector.
The primer can be used only for sequencing inserts cloned into the pTargeT
Vector. The primer sequence is not a binding site for any RNA polymerases and
cannot be used to generate in vitro transcripts.
The sequence of the pTargeT Sequencing Primer is 5´-d(TTACGCCAAGTTAT
TTAGGTGACA)-3´.
The primer is supplied at a concentration of 10ng/μl (1.25pmol/μl) in sterile
water.
Storage Conditions: Store at –20°C.
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