Refolding Record:
| Protein | |
|---|---|
| Protein Name | Hepatitis B virus X antigen |
| Abbreviated Name | HBx |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Hepatitis B virus |
| UniProt Accession | X_HPBV2 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | |
| Domain | None |
| Chimera | None |
| Variants | None |
| Chain Length | 154 |
| Molecular Weight | 16583.0 |
| Pi | 8.63 |
| Molecular Weight | 16583.0 |
| Disulphides | Unknown |
| Full Sequence | |
| Notes | None |
| Expression | |
|---|---|
| Report | Marczinovits I, Somogyi C, Patthy A, Nemeth P, Molnar J. (1997) J Biotechnology, 56, 81-88 |
| Project Aim | Undefined |
| Fusion | N-terminal GST |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21 |
| Expression Temp | 37.0 |
| Expression Time | 2.5h |
| Expression Vector | pGEX-3X |
| Expression Protocol | |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | Lysozyme |
| Pre-Refolding Purification | None |
| Solubility | partial |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | |
| Solubilization Buffer | 8M urea pH 6.5 |
| Refolding Buffer | 4M urea |
| Pre-Refolding Purification | None |
| Tag Cleaved | yes |
| Refolding pH | 6.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | 1mg/ml |
| Refolding Time | 24h |
| Redox Agent | None |
| Redox Agent Concentration | |
| Refolding Protocol | The fusion protein was selectively solubilized from the inclusion bodies by vortexing for 3-5min with 8M urea prepared with deionized distilled water at pH 6.5. The extracted protein was refolded by rapid dilution of the protein solution at 2-3mg/ml in 8M urea with two volumes of deionized distilled water followed by storage at 4 degrees C for 24 h. |
| Refolding Assay | Immunoassay |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | milligram quantities |
| Purity | many contaminating bands on SDS PAGE |
| Notes | |