Cell Signaling pSP-luc+NF Fusion Vector Product pSP-luc+NF Fusion Vector For Research Use Only. Not for Use in Diagnostic Procedures. Description: The pSP-luc+NF Fusion Vector is a luciferase cassette vector containing the engineered firefly luciferase gene, luc+NF. The luc+NF gene is related to the luc+ gene found in the pGL3 family of eukaryotic reporter vectors but has been further modified for maximum flexibility in constructing N-terminal fusions (NF) with luciferase. Subcloning luc+NF into expression vectors provides a useful genetic reporter with exceptional sensitivity. The pSP-luc+NF Fusion Vector is not itself intended for the expression of luciferase in eukaryotic cells, because it does not contain eukaryotic promoters, enhancers or polyadenylation signals. A unique BstEII site has been inserted immediately downstream of the luciferase ATG translation codon, allowing cloned inserts to be positioned immediately downstream of the luc+NF initiation codon. This vector is recommended specifically for applications where N-terminal fusion proteins do not contain an internal ATG codon at the luciferase junction. The luc+NF gene is positioned downstream of an SP6 promoter and a ribosome binding site. An opposing T7 promoter is located immediately downstream of luc+NF. Thus, the pSP-luc+NF Fusion Vector provides a convenient template for the in vitro synthesis of both sense and antisense luciferase transcripts for studies involving in situ hybridization, RNA processing, RNA transfection or coupled in vitro transcription/translation and protein folding. Multiple cloning regions containing recognition sequences for commonly used restriction enzymes are positioned at the 5´ and 3´ ends of luc+NF to provide maximum flexibility in cloning. Luciferase enzymatic activity can be assayed most efficiently using one of the Luciferase Assay Systems. Features: • Flexibility: Multiple cloning regions are positioned at the 5´ and 3´ ends of luc to provide maximum flexibility in cloning. • N-Terminal Fusions with Luciferase: Unique BstEII site located immediately downstream of the luciferase ATG translation codon. Storage Conditions: Store at –20°C. Protocol pSP-luc+NF Fusion Vector Technical Bulletin Part# TB210 Size Cat.# Price (Fr) 20 µg E4471 232.00 pSV-β-Galactosidase Control Vector Product pSV-β-Galactosidase Control Vector For Research Use Only. Not for Use in Diagnostic Procedures. Description: The pSV-β-Galactosidase Control Vector is a positive control vector for monitoring transfection efficiencies of mammalian cells. The SV40 early promoter and enhancer drive transcription of the lacZ gene, which encodes the β-galactosidase enzyme. The pSV-β-Galactosidase Control Vector can be transfected individually or co-transfected with your DNA of interest. β-galactosidase is an excellent reporter enzyme that can be assayed quickly and directly in cell extracts using spectrophotometric, fluorescent or chemiluminescent assays. This reporter enzyme is also widely used for in situ histochemical analysis using the substrate X-Gal. The pSV-β-Galactosidase Control Vector can be co-transfected with your DNA of interest. For example, co-transfection with firefly luciferase gene vectors (pGL3 Vectors) provide cell extracts that can be assayed for both luciferase and β-galactosidase activities. In this manner, the pSV-β-Galactosidase Vector acts as an internal control for transient expression assays. A negative control extract, prepared from mock-transfected cells, should also be assayed for the presence of endogenous β-galactosidase activity in cultured cells. In addition, co-transfection with chloramphenicol acetyltransferase reporter gene vectors (pCAT®3 Vectors) permits assaying for both CAT and β-galactosidase activities. The pSV-β-Galactosidase Vector is a modification of pRSV-β-Gal with SV40 and pUC18 sequences substituted for RSV and pBR322 sequences. The pSV-β-Galactosidase Vector will express β-galactosidase in E. coli due to the presence of the E. coli gpt promoter located upstream of the lacZ gene. Colonies of E. coli containing the pSV-β-Galactosidase Vector will appear blue when plated on media containing X-Gal. Storage Conditions: Store at –20°C. Protocol pSV-ß-Galactosidase Control Vector Technical Bulletin Part# TB094 Size Cat.# Price (Fr) 20 µg E1081 225.00 15 HindIII 414 EcoRI 6815 SV40 Promoter and Enhancer Ampr AlwNI 2372 pSP-luc+NF Fusion Vector (4103bp) HpaI 1777 T7 1726 1721 1716 1708 1702 1700 EcoRV EcoRI XbaI XhoI ClaI BanII 1072 luc+ end NarI 81 Csp45I 217 BclI 628 BbuI 711 SspI 3653 AatII 3771 NdeI 4020 SP6 KpnI NheI BglII AvrII rbs 15 17 24 30 HindIII NcoI BstEII 36 43 45 luc+NF start 47 ori pSV-β-Galactosidase Vector (6820bp) Ampr lacZ EcoRI 3701 PstI 4173 BamHI 4151 SalI 4163 For complete and up-to-date product information visit: www.promega.com/catalog 279 Reporter Assays and Transfection 0752VA08_4A 0325VA03_3A Pagina 282
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