Champion™ pET100 Directional TOPO™ Expression Kit with BL21 Star™ (DE3) One Shot™ Chemically Competent E. coli
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Champion™ pET100 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 canRead more
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Catalog number K10001
Price (USD)/ 20 reactions
1,208.65
Online exclusive
1,212.00 
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Price (USD)/ 20 reactions
1,208.65
Online exclusive
1,212.00 
Save 3.35
Add to cart
Champion™ pET100 Directional TOPO™ Expression Kit with BL21 Star™ (DE3) One Shot™ Chemically Competent E. coli
Catalog numberK10001
Price (USD)/ 20 reactions
1,208.65
Online exclusive
1,212.00 
Save 3.35
-
Add to cart
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 percent 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. coliRNA polymerase and is dedicated to the transcription of your gene of interest. Protein production is further enhanced in the system by the expression strain BL21 Star™ E. coli, which significantly improves the stability of mRNA transcripts and increases protein expression up to ten-fold.

Simplified, efficient Directional TOPO™ cloning allows you to quickly enter a Champion™ pET Expression vector. These kits feature linearized, topoisomerase I-activated Champion™ pET expression vectors for 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
• Greater than 90% of the clones are in the correct orientation for gene expression, reducing the time spent on colony screening

Seven Champion™ pET Directional TOPO™ Expression Vectors are available (Figure 1 and Table 1): Each vector carries a T7lac promoter for high-level expression. Flexible options for simplifying protein detection, cleaving purification tags, selecting plasmid carrying clones, and/or improving protein yields are available.
With the Champion™ pET Directional TOPO™ vectors you can expect highest-level protein production. Figure 2 shows expression of the lacZ gene in Champion™ pET Directional TOPO™ vectors. Figure 3 demonstrates efficient cleavage using TEV protease of the N-terminal tag of a β-galactosidase fusion protein expressed from pET151/D-TOPO™.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Constitutive or Inducible SystemInducible
Inducing AgentIPTG
PromoterT7, lacO
Product TypeTOPO Expression Kit
Selection Agent (Eukaryotic)None
Antibiotic Resistance BacterialAmpicillin (AmpR)
Bacterial or Yeast StrainBL21 Star™(DE3)
CleavageEK (Enterokinase) Recognition Site
Expression SystemE. coli
Expression MechanismCell-Based Expression
Protein TagHis Tag (6x), Xpress Epitope Tag
Cloning MethodDirectional TOPO
Quantity20 reactions
VectorpET
Product LineOne Shot™
Unit Size20 reactions
Contents & Storage
Each Champion™ pET Directional TOPO™ Expression Kit is provided as a complete expression system. The Directional TOPO™ Expression box contains 200 ng of linearized, topoisomerase I-activated Champion™ pET vector; sterile water; dNTPs; 10X PCR Buffer; salt solution; control template and primers; primers for sequencing or PCR screening; and an expression control. Store at -20°C. The One Shot™ TOP10 box contains twenty-one 50-μl aliquots of chemically competent E. coli, S.O.C. medium, and a control plasmid. Store at -80°C. The One Shot™ BL21 Star™(DE3) box contains twenty-one 50-μl aliquots of chemically competent E. coli, S.O.C. medium, and a control plasmid. Store at -80°C. Kits with Lumio™ Technology include 20 μl of the Lumio™ Detection Reagent. Store at -20°C. Guaranteed stable for 6 months when properly stored.