Chaperone plasmid set
The Chaperone Plasmid Set consists of five different plasmids, each of which is designed to express multiple molecular chaperones that function together as a "chaperone team" to enable optimal protein expression and folding and to reduce protein misfolding. Co-expression of a target protein with one of these five chaperone plasmids increases the recovery of soluble protein and minimizes product loss as a result of insoluble protein. Each plasmid carries an origin of replication (ori) derived from pACYC and a chloramphenicol-resistance gene (Cmr) gene, which allows the use of E. coli expression systems containing ColE1-type plasmids that confer ampicillin resistance. The chaperone genes are situated downstream of the araB orPzt-1 (tet) promoters; as a result, expression of target proteins and chaperones can be individually induced if the target gene is placed under the control of different promoters (e.g., lac). These plasmids also contain the necessary regulator (araC or tetr) for each promoter.
The Chaperone Plasmid Set consists of five different plasmids, each of which is designed to express multiple molecular chaperones that function together as a "chaperone team" to enable optimal protein expression and folding and to reduce protein misfolding. Co-expression of a target protein with one of these five chaperone plasmids increases the recovery of soluble protein and minimizes product loss as a result of insoluble protein. Each plasmid carries an origin of replication (ori) derived from pACYC and a chloramphenicol-resistance gene (Cmr) gene, which allows the use of E. coli expression systems containing ColE1-type plasmids that confer ampicillin resistance. The chaperone genes are situated downstream of the araB orPzt-1 (tet) promoters; as a result, expression of target proteins and chaperones can be individually induced if the target gene is placed under the control of different promoters (e.g., lac). These plasmids also contain the necessary regulator (araC or tetr) for each promoter.
Compatible E. coli expression systems
- The Chaperone Plasmid Set offers a pACYC ori and Cmr gene, allowing the use of E. coli expression systems that utilize colE1-type plasmids with ampicillin resistance. The Chaperone Plasmid Set cannot be used in combination with chloramphenicol-resistant E. coli host strains or expression plasmids that carry a chloramphenicol-resistance gene. For example, E. coli BL21(DE3), which is often used with pET and other systems, is a compatible host strain. However, E. coli BL21(DE3) pLysS and BL21(DE3) pLysE, both of which contain the pACYC replication origin and Cmr gene-containing pLysS or pLysE plasmids, cannot be used with this system.
- This product prescribes a two-step method to construct target protein/chaperone co-expression systems, the first step describing how to prepare an E. coli host transformed with just the chaperone plasmid, and the second step describing how to prepare competent cells from the chaperone plasmid-containing strain and to transform this strain with the target protein-containing plasmid.
Plasmids in the Chaperone Plasmid Set work well in combination with the pCold Expression System Vectors.
More Information
Applications
- Correct in vivo folding of expressed recombinant proteins in E. coli
- Reduction of insoluble protein during expression
Components
| Plasmid pG-KJE8 (10 ng/µl) | 100 µl |
| Plasmid pGro7 (10 ng/µl) | 100 µl |
| Plasmid pKJE7 (10 ng/µl) | 100 µl |
| Plasmid pGTf2 (10 ng/µl) | 100 µl |
| Plasmid pTf16 (10 ng/µl) | 100 µl |
Product citations
Nishihara, K., Kanemori, M., Kitagawa, M., Yanagi, H. & Yura, T. Chaperone coexpression plasmids: differential and synergistic roles of DnaK-DnaJ-GrpE and GroEL-GroES in assisting folding of an allergen of Japanese cedar pollen, Cryj2, in Escherichia coli. Appl. Environ. Microbiol. 64, 1694–9 (1998).
Nishihara, K., Kanemori, M., Yanagi, H. & Yura, T. Overexpression of trigger factor prevents aggregation of recombinant proteins in Escherichia coli. Appl. Environ. Microbiol. 66, 884-9 (2000).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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