Invitrogen™ Champion™ pET101 Directional TOPO™ Expression Kit with BL21 Star™ (DE3) One Shot™ Chemically Competent E. coli

Includes

200ng 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. The TOP10 box contains twenty-one 50μL aliquots of chemically competent E. coli, S.O.C. medium, and a control plasmid.

Yields protein of interest at levels greater than 50 percent of total cellular protein in E. coli

• Based on T7 expression vectors originally developed by Studier and colleagues, high-level expression is achieved because T7 RNA polymerase is more processive than native E. coliRNA polymerase and is dedicated to transcription of gene of interest
• Protein production is further enhanced in system by expression strain BL21 StarE. coli, which significantly improves stability of mRNA transcripts and increases protein expression up to ten-fold
• Simplified, efficient Directional TOPO cloning allows to quickly enter ChampionpET Expression vector
• Features linearized, topoisomerase I-activated ChampionpET expression vectors for five-minute directional cloning
• Directional TOPO Cloning technology facilitates gene expression becausa proofreading enzyme is used for PCR, resulting in fewer errors in cloned genes
• Greater than 90% of clones are in correct orientation for gene expression, reducing time spent on colony screening

Bacterial Expression, Protein Expression, Proteins, Expression, Isolation and Analysis

Constitutive or Inducible System

Inducible

Inducing Agent

IPTG

Promoter

T7, lacO

Product Type

TOPO Expression Kit

Selection Agent (Eukaryotic)

None

Content And 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.

Antibiotic Resistance Bacterial

Ampicillin (AmpR)

Bacterial or Yeast Strain

BL21 Star™(DE3)

Expression System

E. coli

Expression Mechanism

Cell-Based Expression

Protein Tag

His Tag (6x), V5 Epitope Tag

Cloning Method

Directional TOPO

Quantity

20 reactions

Vector

pET

Product Line

One Shot™

For Research Use Only. Not for use in diagnostic procedures.