Thermo Fisher Scientific Champion pET200 Directional TOPO Expression Kit with BL21 Star (DE3) One Shot Chemically Competent E. coli

Invitrogen™ Champion™ pET200 Directional TOPO™ Expression Kit with BL21 Star™ (DE3) One Shot™ Chemically Competent E. coli

The Champion™ pET Expression System yields the highest-level protein production in E. coli. During expression, your protein of interest can reach levels greater than 50% of total cellular protein.
Based on T7 expression vectors originally developed by Studier and colleagues (1–3), high-level expression is achieved because the T7 RNA polymerase is more processive than native E. coli RNA polymerase and is dedicated to the transcription of your gene of interest.

Protein production is further enhanced by the BL21 Star™ E. coli strain, which improves mRNA transcript stability and increases protein expression up to ten-fold.

Directional TOPO™ Cloning Advantages

Simplified, efficient Directional TOPO™ cloning allows rapid entry into a Champion™ pET Expression vector. These kits feature linearized, topoisomerase I-activated Champion™ pET expression vectors enabling five-minute directional cloning.

Directional TOPO™ Cloning technology facilitates gene expression because:

• A proofreading enzyme is used for PCR, resulting in fewer errors in cloned genes
• Over 90% of clones are in the correct orientation for expression, reducing colony screening time

Available Vectors

Seven Champion™ pET Directional TOPO™ Expression Vectors are available, each with a T7_lac_ promoter for high-level expression.
Flexible options are provided for:

• Simplifying protein detection
• Cleaving purification tags
• Selecting plasmid-carrying clones
• Improving protein yields

With the Champion™ pET Directional TOPO™ vectors, you can expect high-level protein production.
Figure 2 shows expression of the lacZ gene, and Figure 3 demonstrates efficient cleavage using TEV protease of the N-terminal tag of a β-galactosidase fusion protein expressed from pET151/D-TOPO™.

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