Refolding Record:
| Protein | |
|---|---|
| Protein Name | Phage lambda terminase enzyme |
| Abbreviated Name | phage lambda |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Bacteriophage lambda |
| UniProt Accession | P03707 |
| 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 | 184 |
| Molecular Weight | 20441.1 |
| Pi | 4.97213 |
| Molecular Weight | 20441.1 |
| Disulphides | 0 |
| Full Sequence |
MEVNKKQLADIFGASIRTIQNWQEQGMPVLRGGGKGNEVLYDSAAVIKWYAERDAEIENE
KLRREVEELRQASEADLQPGTIEYERHRLTRAQADAQELKNARDSAEVVETAFCTFVLSR
IAGEIASILDGLPLSVQRRFPELENRHVDFLKRDIIKAMNKAAALDELIPGLLSEYIEQS
G
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Meyer JD, Hanagan A, Manning MC, Catalano CE. Meyer JD, Hanagan A, Manning MC, Catalano CE. (1998) International J. Biological Macromolecul, 23, 27-36 |
| Project Aim | Undefined |
| Fusion | None |
| 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 | 2h |
| Expression Vector | pKKT7(-H) |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution/Dialysis combination |
| Wash Buffer | PBS, 5mM EDTA, 25% sucrose, 1% Triton X-100 |
| Solubilization Buffer | 50mM Tris-HCL, 5mM EDTA, 6M guanidinium chloride, pH 8.0 |
| Refolding Buffer | 50mM Tris-HCL, 5mM EDTA |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | 48h |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The partially purified gpNu1-containing pelletwas taken into 150 ml buffer A (50 mM Tris,pH 8.0, 5 mM EDTA) containing 6 M guanidinium hydrochloride, briefly sonicated and gently stirred overnight. Insoluble material wasremoved by centrifugation (14K×g?5 min),the sample was diluted to 4 l with buffer Acontaining 4 M GDN and 100 mM NaCl andthe protein was refolded by dialysis against 35 L buffer A for 48 h with one buffer change. |
| Refolding Assay | Enzyme activity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | >70% |
| Notes | |