For parallel expression of His-tagged proteins in E. coli, mammalian cells, and baculovirus-infected insect cells using a single construct

  • No need for time-consuming subcloning procedures
  • Obtain post-translational modifications in insect or mammalian cells
  • One construct provides efficient expression in three expression systems
The pQE TriSystem Vector allows for high-level expression of His-tagged proteins from a single vector containing three different expression systems. There is the T5 promoter/lac operator transcription–translation system for expression in E. coli, the p10 promoter for baculovirus-based expression in insect cells, and the CAG (CMV/actin/globin) promoter for expression in mammalian cells.
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pQE-TriSystem Vector
25 μg pQE-TriSystem Vector DNA
33903
$411.00
$0.00
The pQE-TriSystem Vector is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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pQE Vectors.|pQE TriSystem.|
Numbered elements listed in table.||
Performance
The pQE-TriSystem vector contains CAG, T5, and p10 promoters that enable 6xHis-tagged protein expression in mammalian, E. coli, and baculovirus-infected insect cells, respectively (see figure pQE TriSystem). Preliminary studies can be carried out in bacterial expression systems, using the strong T5 promoter, which is recognized by E. coli polymerase, and allows efficient expression of proteins in any E. coli strain. If expression in mammalian or insect cells is required — to obtain post-translational modifications, for example — the same construct can be used without the need for time-consuming subcloning procedures.
Principle

QIAexpress pQE vectors combine a powerful phage T5 promoter (recognized by E. coli RNA polymerase) with a double lac operator repression module to provide tightly regulated, high-level expression of recombinant proteins in E. coli. Protein synthesis is effectively blocked in the presence of high levels of lac repressor and the stability of cytotoxic constructs is enhanced. The pQE vectors (see table and figure pQE Vectors) enable placement of the 6xHis tag at either the N- or C-terminus of the recombinant protein.

Elements present in QIAexpress pQE Vectors
Element Description
1. Optimized promoter/operator element Consists of the phage T5 promoter and two lac operator sequences, which increase the probability of lac repressor binding and ensure efficient repression of the powerful T5 promoter
2. Synthetic ribosomal binding site RBSII For efficient translation
3. His-tag coding sequence Either 5' or 3' to the polylinker cloning region
4. Translational stop codons In all reading frames for convenient preparation of expression constructs
5. Two strong transcriptional terminators t0 from phage lambda, and T1 from the rrnB operon of E. coli, to prevent read-through transcription and ensure stability of the expression construct

6. ColE1 origin of replicatio

From pBR322
7. beta-lactamase gene (bla) Confers ampicillin resistance
Procedure

Inserts encoding proteins of interest are cloned into appropriate constructs and transformed into a suitable E. coli strain for expression. Expression is induced by addition of IPTG. Vector pQE-TriSystem constructs can be transformed into E. coli, used as a shuttle vector for recombinant protein expression in insect cells, or transfected into mammalian cells.

Applications

The QIAexpress Expression System provides high-level expression of proteins suitable for
many applications, including:

Purification of functional, conformationally active proteins
Purification under denaturing conditions for antibody production
Crystallization for determination of three-dimensional structure
Assays involving protein-protein and protein-DNA interactions
Feature
Specifications
All three reading frames provided No
Expression In vivo
Expression species E.coli, mammalian & insect cells
In-frame cloning necessary Yes
N- or C-terminal tag C-terminal tag
Tag 6xHis tag
Tag removal sequence No

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Kit Handbooks
1
A handbook for high-level expression and purification of 6xHis-tagged proteins
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