Science Catalog 2012 Life Cell Signaling pAdVAntage™ Vector Product pAdVAntage™ Vector 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 pAdVAntage™ Vector Technical Bulletin Part# TB207 Size Cat.# Price (Fr) 20 µg E1711 229.00 pSI Mammalian Expression Vector Product pSI Mammalian Expression Vector For Research Use Only. Not for Use in Diagnostic Procedures. 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 pCI and pSI Mammalian Expression Vectors Technical Bulletin BglII ori SV40 Enhancer/ Early Promoter Ampr Vector (3632bp) pSI Intron SV40 Late poly(A) f1 ori BamHI PstI Bst98I Bst98I T7 NheI XhoI EcoRI MluI XbaI SalI AccI SmaI BstZI NotI 678 684 689 695 707 713 714 720 724 724 Part# TB206 Size Cat.# Price (Fr) 20 µg E1721 583.00 302 For complete and up-to-date product information visit: www.promega.com/catalog ➞ 0684VA06_4B Pagina 305
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