规格
Antibiotic Resistance Bacterial:	Ampicillin (AmpR)
Bacterial or Yeast Strain:	TOP10
Cleavage:	EK (Enterokinase) Recognition Site
Cloning Method:	TOPO®-TA
Expression Mechanism:	Cell-Based Expression
Expression System:	E. coli
Product Line:	TOPO®, ThioFusion™
Constitutive or Inducible System:	Inducible
Inducing Agent:	Arabinose
Promoter:	araBAD
Selection Agent (Eukaryotic):	None
Vector:	pBAD
Protein Tag:	His Tag (6x), V5 Epitope Tag, Thioredoxin

储存

Each pBAD/TOPO® ThioFusion™ Expression Kit contains two boxes and the LMG194 E. coli stab. The pBAD/TOPO® ThioFusion™ Expression box contains all of the reagents necessary for PCR (except Taq polymerase) including 200 ng of topoisomerase I-activated pBAD/Thio-TOPO® vector, sterile water, dNTPs, 10X PCR buffer, salt solution, control template and primers, 20% L-arabinose, primers for sequencing or PCR screening, and an expression control plasmid. Store at -20°C. The One Shot® box contains all of the reagents necessary for transformation including 21 single-use 50 µl aliquots of chemically competent TOP10 E. coli, S.O.C. medium, and pUC19 supercoiled control plasmid. Store the One Shot® box at -80°C. Store the LMG194 E. coli stab at 2 - 8°C. All components are guaranteed stable for 6 months when properly stored.

描述

The pBAD/TOPO® ThioFusion™ Expression Kit is designed for one-step cloning and regulated expression of thioredoxin fusion proteins in E. coli(Figure 1). The pBAD/Thio-TOPO® vector encodes His-Patch thioredoxin as an N-terminal fusion partner. The thioredoxin fusion can significantly increase the solubility of many difficult-to-express proteins and improve the yield of protein production. The vector includes the following additional features (Figure 2):

• The araBAD promoter for tightly regulated expression in E. coli I-activated vector for 5-minute TOPO® Cloning of Taq-amplified PCR products
• C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• C-terminal V5 epitope for detection with an Anti-V5 Antibody
• Enterokinase cleavage site for efficient cleavage of N-terminal fusion tag with EKMax™ Enterokinase

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