pTrcHis TOPO® TA Expression Kit

Catalog number: K441001

Invitrogen™  Related applications: Protein Expression

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The pTrcHis- and pTrcHis2-TOPO® TA Expression Kits allow fast, efficient cloning of PCR products into a prokaryotic expression vector for high-level, regulated expression from the trc promoter. The vector in each kit, pTrcHis-TOPO® or pTrcHis2-TOPO®, includes a minicistron element for enhanced translation efficiency of eukaryotic proteins in E. coli. Both vectors are provided linearized and activated with topoisomerase I for 5-minute TOPO® Cloning with >85% cloning efficiency. To simplify protein purification and detection, the pTrcHis-TOPO® and pTrcHis2-TOPO® vectors include the following features:


N-terminal Xpress™ epitope for detection with an Anti-Xpress™ Antibody
• N-terminal polyhistidine (6xHis) tag for purification using nickelchelating resin and detection with an Anti-HisG Antibody
• Enterokinase cleavage site for efficient removal of N-terminal fusion tags

C-terminal c-myc epitope for detection with an Anti-myc Antibody
• C-terminal polyhistidine (6xHis) tag for purification using nickel-chelating resin and detection with an Anti-His(C-term) Antibody
For Research Use Only. Not for use in diagnostic procedures.


Expression: E. coli
Promoter: Trc and lacO
Bacterial or Yeast Strain: TOP10
Cleavage: EK (Enterokinase) Recognition Site
Vector Type: pTrc
Product Size: 20 reactions
Cloning Method: TOPO®-TA
Inducing Agent: IPTG
Expression Mechanism: Cell-Based Expression
Protein Tag or Fusion: His Tag (6x), Xpress Epitope Tag
Selection Agent (Eukaryotic): None
Constitutive or Inducible System: Inducible
Antibiotic Resistance (Bacterial): Ampicillin (AmpR)

Contents & storage

The pTrcHis- and pTrcHis2-TOPO® TA Expression Kits each contain two boxes. The TOPO® TA box contains 200 ng of linearized, topoisomerase I-activated pTrcHis-TOPO® or pTrcHis2-TOPO® vector, sterile water, dNTPs, 10X PCR buffer, a control template and primers, primers for sequencing or PCR screening, and an expression control plasmid. The One Shot® TOP10 box contains twenty-one, 50-µl aliquots of chemically competent TOP10 E. coli, S.O.C. medium, and a supercoiled control plasmid. Store the TOPO TA Cloning® box at -20°C. Store the One Shot® box at -80°C. All reagents are guaranteed stable for 6 months when properly stored.


Manuals & protocols

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