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Abstract for pTARGET™ Mammalian Expression System
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 cutting the pTARGET™
Vector with EcoR V and adding a 3´ terminal thymidine to both ends. 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; secondly 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, allowing blue/white recombinant
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. |
