70942  pET NusA Fusion System 43.1

Production of soluble, active proteins in E. coli

pET Expression Systems and pET
pET Expression Systems and pET Expression Systems plus Competent Cells provide core reagents needed for target gene cloning and expression.

Components: pET Expression Systems
Components for pET Expression Systems are similar for all systems unless otherwise stated with the specific pET Expression System description. pET Expression Systems include:
• 10 µg pET vector DNA (for each indicated plasmid)
• 0.2 ml BL21 Glycerol Stock
• 0.2 ml BL21(DE3) Glycerol Stock
• 0.2 ml BL21(DE3)pLysS Glycerol Stock
• 0.2 ml Induction Control Glycerol Stock

Components: pET Expression Systems plus Competent Cells
pET Expression Systems plus Competent Cells contain all of the components of the specific pET Expression System, as well as the following additional components, unless otherwise stated with the specific pET Expression System description:
• 0.2 ml NovaBlue Competent Cells
• 0.2 ml BL21(DE3) Competent Cells
• 0.2 ml BL21(DE3)pLysS Competent Cells
• 2 × 0.2 ml SOC Medium
• 10 µl Test Plasmid

These components are sufficient for up to 10 transformations in each host.

Purification and Detection Reagents
Purification and detection reagents are available separately. For complete product descriptions and ordering information, refer to the Protein Purification and Antibodies, Conjugates & Detection Tools chapters.

pET NusA Fusion System 43.1
The pET NusA Fusion Systems are designed for cloning and high-level expression of polypeptide sequences fused with the 495-aa NusA (Nus•Tag) protein. The NusA protein has been identified as having the highest potential for solubility when a database of more than 4000 E. coli proteins was subjected to solubility modeling (Harrison 2000, Davis 1998). Greater than 85% of the expressed protein was soluble in tests with each of four different NusA fusion proteins (Harrison 2000).

Another pET vector with the Nus•Tag sequence is pET-50b(+) (Cat. No. 71471-3). The pET-43.1 vectors incorporate the Nus•Tag fusion partner and are also compatible with the trxB/gor mutant Origami, Origami 2, Origami B, Rosetta-gami, Rosetta-gami 2, and Rosetta-gami B strains, which facilitatedisulfide bond formation in the cytoplasm (Davis 1998). Disulfide-bonded proteins maybe obtained by employing the combination of a pET-43.1 or pET-44 vector and these trxB/gor host strains. Unfused proteins can be produced by using the Nde I cloning site.

This product is covered under license from Brookhaven National Labs. Commercial entities need to obtain a research use license prior to purchase. Please contact your licensing department to confirm that your company already holds a research use license.