Principle

The Cherry tag vectors allow direct visualization of your protein of interest during the whole process of protein
production and purification without the need for any special requirements or reagents. When using the Cherry tag vectors, your gene of interest is fused to a small sequence encoding a red polypeptide (heme binding part of cytochrome, 11 kDa) providing a visual aid for estimating expression level and solubility: bacteria expressing the fusion protein are red when the fused protein is soluble. The red color also constitutes a visual marker throughout protein purification. Concentration of the fusion protein can be determined easily by spectral measurement (before and after purification). In addition, the Cherry tag is highly soluble and thus, can increase the solubility of target proteins (for more information about protein solubility using Cherry^TM kits, consult this application note). When using the Cherry tag as a N-terminal tag, it can be cleaved after purification using enterokinase (a recognition site is inserted at the C-terminal end of the tag sequence). The vector of the Cherry^TMCodon kit (pSCherry2) encodes also rare t-RNAS allowing high-level expression of heterologous proteins (for more information, see the Staby^TMCodon kit). Moreover, this vector is perfectly stabilized with or without antibiotics due to the action of the Staby^TM technology.

The Cherry^TMCodon system is compatible with the use of an auto-inducible medium as Staby^TMSwitch.

Kit Contents:

 pSCherry2 vector for Cherry N-terminal tag

pSCherry4 vector for Cherry C-terminal tag

Benefits:

• Direct visualization of the protein of interest
• No need of any special apparatus or reagents
• Rapid screening and optimization of proteinexpression and solubility
• Easy quantification of the protein of interest
• Efficient expression of heterologuous genes due to Codon Correction
• High yield of protein production using StabyTM systems with or without antibiotics
• Not necessary to mutate each rare codon;
• Reduced background of “parasite proteins”
• No additional plasmid in the bacteria