Refolding Record:
| Protein | |
|---|---|
| Protein Name | Culture Filtrate Protein 10 |
| Abbreviated Name | CFP-10 |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Mycobacterium tuberculosis |
| UniProt Accession | P0A566 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 117 |
| Molecular Weight | 12717.8 |
| Pi | 6.03 |
| Molecular Weight | 12717.8 |
| Disulphides | 0 |
| Full Sequence |
HHHHHHHSSGLVPRGSHMAEMKTDAATLGQEAGNFERISGDLKTQIDQVESTAGSLQGQWRGAAGTAAQAAVVRFQEAANKQKQELDEISTNIRQAGVQYSRADEEQQQALSSQMGF
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Berthet, F. X., Rasmussen, P. B., Rosenkrands, I., Andersen, P., Gicquel, B. (1998) Microbiology, 144, 3195-3203 |
| Project Aim | Structure-Function |
| Fusion | N-terminal poly-histidine tag |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | None |
| Expression Temp | 37.0 |
| Expression Time | not stated |
| Expression Vector | pET-14b |
| Expression Protocol | No protocol provided |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | Size-exclusion chromatography |
| Solubility | not stated |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | n/a |
| Solubilization Buffer | 8 M urea |
| Refolding Buffer | 25mM HEPES, pH 8.5 |
| Pre-Refolding Purification | Size-exclusion chromatography |
| Tag Cleaved | no |
| Refolding pH | 8.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | Not specified |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Recombinant CFP-10 was purified using Talon resin after solublization in solublization buffer. Fractions containing CFP-10, determined by SDS-PAGE, were pooled and applied to a Resource Q column equilibrated with 10mM TrisHCl, 3M urea, pH 7.5. and eluted using a linear gradient (0-1M) NaCl. Fractions containing CFP-10 were pooled and dialysed against refolding buffer, concentrated by dialysis against PEG 20000 and then again against refolding buffer. |
| Refolding Assay | SDS-PAGE,Sequence Analysis |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |