2012 Promega catalogue
2012 Promega catalogue
2012 Promega catalogue
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Cell Signaling<br />
pRL Renilla Luciferase Control Reporter<br />
Vectors<br />
Product Size Cat.# Price ($)<br />
pRL-SV40 Vector 20 µg E2231 218.00<br />
pRL-TK Vector 20 µg E2241 218.00<br />
pRL-CMV Vector 20 µg E2261 218.00<br />
pRL-null Vector 20 µg E2271 218.00<br />
For Research Use Only. Not for Use in Diagnostic Procedures.<br />
Description: The pRL Vectors are wildtype Renilla luciferase (Rluc) control<br />
reporter vectors. The pRL Vectors, which provide constitutive expression of<br />
Renilla luciferase, can be used in combination with a firefly luciferase vector<br />
to cotransfect mammalian cells. Expression of Renilla luciferase provides an<br />
internal control value to which expression of the experimental firefly luciferase<br />
reporter gene may be normalized. The pRL Vectors contain the cDNA<br />
encoding Renilla luciferase (Rluc) cloned from the anthozoan coelenterate<br />
Renilla reniformis (sea pansy). Four different promoter configurations are available.<br />
The HSV-thymidine kinase promoter (pRL-TK) is relatively weak and may<br />
be particularly useful in providing neutral constitutive expression of the Renilla<br />
luciferase control reporter. The early SV40 enhancer/promoter region (pRL-<br />
SV40) and the CMV immediate early enhancer/promoter region (pRL-CMV)<br />
typically provide high-level transcription and, therefore, may be less suitable for<br />
co-reporter applications involving experimental vectors with robust regulatory<br />
elements. In general, we recommend validating the performance of specific<br />
co-reporter combinations in the desired target cells. In addition to the modified<br />
Rluc reporter gene, all pRL Vectors are isolated from a dam–/dcm–<br />
E. coli K host strain, allowing digestion with restriction enzymes that are<br />
sensitive to dam and dcm methylation.<br />
Features:<br />
• A T7 promoter is located immediately upstream of Rluc, allowing in vitro<br />
synthesis of Renilla luciferase.<br />
• The SV40 late poly(A) signal sequence is positioned downstream of Rluc to<br />
provide efficient transcription termination and mRNA polyadenylation.<br />
• A prokaryotic origin of replication and β-lactamase gene allow selected<br />
propagation of the pRL vectors in E. coli host strains.<br />
• To avoid DNA methylation, all pRL Vectors are isolated from a dam–/dcm–<br />
E. coli K host strain.<br />
Storage Conditions: Store vectors at –20°C.<br />
Protocol Part#<br />
pRL Renilla Luciferase Reporter Vectors Technical Bulletin TB550<br />
BamHI<br />
1499<br />
SV40 late<br />
poly(A)<br />
1247 XbaI<br />
Amp r<br />
pRL-null<br />
Vector<br />
(3320bp)<br />
Rluc<br />
ori<br />
T7<br />
Promoter<br />
NheI 299<br />
Csp45I 315<br />
BglII<br />
XhoI<br />
SacI<br />
EcoICR I<br />
HindIII<br />
NdeI<br />
NsiI<br />
SphI<br />
SpeI<br />
NarI<br />
SalI<br />
MluI<br />
EcoRI<br />
XmaI<br />
SmaI<br />
PstI<br />
For complete and up-to-date product information visit: www.promega.com/catalog<br />
1<br />
5<br />
12<br />
10<br />
15<br />
24<br />
30<br />
32<br />
34<br />
41<br />
47<br />
51<br />
58<br />
65<br />
67<br />
77<br />
1352VA01_6A<br />
pGL3 Luciferase Reporter Vectors<br />
Product Size Cat.# Price ($)<br />
pGL3-Basic Vector 20 µg E1751 214.00<br />
pGL3-Control Vector 20 µg E1741 214.00<br />
pGL3-Enhancer Vector 20 µg E1771 214.00<br />
pGL3-Promoter Vector<br />
For Research Use Only. Not for Use in Diagnostic Procedures.<br />
20 µg E1761 214.00<br />
Description: The pGL3 Luciferase Reporter Vectors provide a basis for the<br />
quantitative analysis of factors that potentially regulate mammalian gene<br />
expression. These may be cis- or trans-acting factors. The backbone of the<br />
pGL2 Luciferase Reporter Vectors was redesigned for the pGL3 Vectors for<br />
increased expression, with a modified coding region for firefly (Photinus pyralis)<br />
luciferase that has been optimized for monitoring transcriptional activity<br />
in transfected eukaryotic cells. The assay of this genetic reporter is rapid,<br />
sensitive and quantitative. In addition, the Luciferase Reporter Vectors contain<br />
numerous features aiding in the structural characterization of the putative<br />
regulatory sequences under investigation.<br />
For the most advanced reporter vectors and widest selection of features, please<br />
see the pGL4 Luciferase Reporter Vectors.<br />
Features:<br />
• Easy to Use: NcoI site located at 5´ end of luc+ gene allows creation of<br />
fusions with reporter gene using a unique NcoI site.<br />
• Flexible: Placement of SmaI site in the MCS allows blunt-ended inserts<br />
to be ligated into the MCS and restricted on either side by other restriction<br />
endonucleases.<br />
• Versatile: XbaI site just downstream of luc+ gene facilitates insertions into<br />
the 3´ untranslated region of mRNA or subcloning of the luciferase gene.<br />
Storage Conditions: Store vectors at –20°C.<br />
Protocol Part#<br />
pGL3 Luciferase Reporter Vectors Technical Manual TM033<br />
2448 SalI<br />
2442 BamHI<br />
SV40 Enhancer<br />
SV40 late<br />
poly(A) signal<br />
(for luc+ reporter)<br />
HpaI 2094<br />
ori<br />
XbaI 1934<br />
Amp r<br />
pGL3-Control<br />
Vector<br />
(5256bp)<br />
f1 ori<br />
luc+<br />
Synthetic poly(A)<br />
signal / transcriptional<br />
pause site<br />
(for background<br />
reduction)<br />
KpnI<br />
SacI<br />
MluI<br />
NheI<br />
SmaI<br />
XhoI<br />
BgIII<br />
NcoI 278<br />
NarI 313<br />
5<br />
11<br />
15<br />
21<br />
28<br />
32<br />
36<br />
SV40 Promoter<br />
HindIII 245<br />
0747VA08_4A<br />
273<br />
15<br />
Reporter Assays and Transfection<br />
Section<br />
Contents<br />
Table of<br />
Contents
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