Refolding Record:
| Protein | |
|---|---|
| Protein Name | Ribosomal protein P1-like protein |
| Abbreviated Name | Ribosomal protein P1-like |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Leishmania donovani |
| UniProt Accession | Q8I867 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | n |
| Domain | n/a |
| Chimera | Beta-galactosidase - P1 like fusion protein |
| Variants | n/a |
| Chain Length | 156 |
| Molecular Weight | 15459.6 |
| Pi | 4.64 |
| Molecular Weight | 15459.6 |
| Disulphides | 0 |
| Full Sequence |
MITPSSKLTLTKGNKSWSSTAVAAALELVDPPGCRNMSAETLACTYAALMLSDAGLPTSAENIAAAVKAAGVEMRPTLPIIFARFLEKKSVETLMAAAAAQAPTAAXAPSPAAGAASAAXXGGKVEDKKKDEPEEEGDDDMGFGLFDGGPGTQFAL
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Arora, S., Pal, N. S., Mujtaba, S. (2005) Experimental Parasitology, 109, 163-170 |
| Project Aim | Antigenic Determinant Identification |
| Fusion | Beta-galactosidase |
| 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 | n/a |
| Expression Vector | pBluescript SK+ |
| Expression Protocol | Cells were centrifuged and pellet re-suspended in 20 mM TrisHCl, pH 7.5, 20% sucrose, and 1 mM EDTA. |
| Method of Induction | IPTG |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | Osmotic shock + sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | Tris-EDTA buffer, pH 8.0, 5 M guanidinium hydrochloride, 5 mM EDTA |
| Refolding Buffer | 20 mM Tris-Cl buffer, pH 8.0, glycerol, DTT, PMSF |
| Pre-Refolding Purification | None |
| Tag Cleaved | no |
| Refolding pH | 8.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | DTT |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Inclusion bodies were re-suspended in solubilization buffer, centrifuged and sonicated. They were then refolded in refolding buffer. |
| Refolding Assay | Immunoassay,Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | Glycerol |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |