pcDNA™3.3 TOPO® Mammalian Expression Vector

TOPO® Cloning

Powerful mammalian expression vector delivers 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

Invitrogen's 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 2).
Express Multimilligram Level
Figure 2. 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.
Figure 3. 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 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


pcDNA TOPO Vector

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 the best 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.
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