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.
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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.

Ordering Information

Catalog # Name Size List Price (USD) Qty
K830001 pcDNA™3.3-TOPO® TA Cloning® Kit 20 reactions 960.00