Constitutive-Mammalian-Protein-Expression
Powerful mammalian expression vector is designed to deliver high protein yields

The pcDNA™3.3-TOPO® TA Mammalian Expression Vector enables expression of exceptionally high levels of recombinant protein in adherent-or suspension-adapted mammalian cells.

  • Obtain up to five-fold higher protein yields
  • Fast, simple, high efficient TOPO® cloning saves time
  • Express native proteins, ideal for antibody production and structural biology
  • Ideal for transient or stable mammalian expression
  • Complete kit contains all the necessary cloning reagents

Ideal for large-scale protein production

The pcDNA™3.3-TOPO® vector contains a modified, enhanced CMV promoter that enables extremely high protein expression. The vector is ideal for large-scale protein production, especially when used in combination with our FreeStyle™ MAX CHO or 293 Expression Systems. We expressed several proteins and observed 8–30 mg/L of recombinant protein produced using our FreeStyle™ systems (Figure 1).

High-yields-recombinant-protein

Figure 1. High yields of recombinant protein. We used pcDNA™3.3-TOPO® vector to clone and express milligram levels of erythropoietin (EPO), Factor IX (FIX), and IgG in FreeStyle™ CHO-S cells. pcDNA™3.3-TOPO® constructs containing the relevant PCR-derived gene sequences were transfected into FreeStyle™ CHO-S cells in 30 mL volumes. Heavy and light chain genes were co-transfected for IgG expression. Four to six days posttransfection, EPO and FIX were quantified using immobilized goat anti–human IgG antibody. Each bar shows the average of two expression runs.

pcDNA3-3vector-map

Figure 2. pcDNA™3.3 vector map. The latest version in the pcDNA vector family, pcDNA™3.3 offers you the flexibility to express a native protein or add your favorite purification tag to the gene of interest prior to TOPO® cloning.

More free time and better cloning efficiency in just three simple steps

Our pcDNA™3.3 vector incorporates TOPO® cloning technology, which removes time-intensive and unreliable steps from your cloning workflow, allowing you to perform bench-top cloning reactions in typically just 5 minutes. With up to 95% recovery of your desired clone, you always have the clone you need for downstream experiments.

Five-fold better expression
Five-fold-better-expression

Figure 3.We cloned and measured expression of luciferase (panel A) and ß-galactosidase (panel B) in pcDNA™3.3-TOPO® vector, pCI and pCMV-Script® vectors. After transient transfection into adherent GripTite™ 293 MSR cells, ß-galactosidase activity and luciferase activity is 2- and 5-fold higher, respectively, using pcDNA™3.3 vector compared to competitor vectors. Error bars shown represent standard deviation (n = 6).

Complete kit

The pcDNA™3.3 TOPO® TA Cloning Kit includes high-efficiency One Shot® TOP10 Competent Cells for superior cloning performance. Our OptiCHO™ Antibody Express Kit, which includes the pcDNA™3.3 TOPO® TA Cloning Kit, enables stable cell line creation and antibody production in a serum-free dihydrofolate reductase-deficient (DHFR–) Chinese hamster ovary (CHO) DG44 cell line.