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Mammalian Expression Vector for Efficient Cloning of PCR Fragments

rect folding and posttranslational modifications of the gene product. Such posttranslational modifications are often required for the activity of eukaryotic proteins. To make the PCR-Script cloning strategy available to researchers interested in expressing their genes in eukaryotic cells, Stratagene has created the pCMV-Script PCR cloning kit, a mammalian version of the PCR-Script cloning system.

Figure 1

The pcmv-script PCR cloning kit features the pCMV-Script vector (figure 1), a mammalian expression vector that uses the cytomegalovirus (CMV) promoter for constitutive expression in a wide variety of cell lines. The pCMV-Script vector shares many essential characteristics with the pPCR-Script vector: capacity to clone inserts generated with any PCR enzyme, high efficiency and 1-hour ligation. However, the pCMV-Script vector has many distinguishing features. The powerful CMV promoter and the SV40 polyadenylation site allow high-level constitutive expression in a wide variety of cell lines. In addition, the neomycin-resistance gene is under control of the prokaryotic b-lactamase promoter to provide kanamycin resistance in bacteria as well as the SV40 early promoter to provide G418 resistance in mammalian cells.

The pCMV-Script PCR cloning kit provides reagents for 25 reactions. Each kit contains predigested pCMV-Script vector, cloned Pfu DNA polymerase for polishing,3 Srf I restriction enzyme, T4 DNA ligase and Epicurian Coli XL10-Gold ultracompetent cells. The kit also includes a control PCR insert and the necessary cloning and polishing buffers.

Cloning PCR fragments

The cloning efficiency of the pCMV-Scrip
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