Refolding Record:
| Protein | |
|---|---|
| Protein Name | Sj Serpin |
| Abbreviated Name | Sj Serpin |
| SCOP Family | Serpins |
| Structure Notes | |
| Organism | Schistosoma japonicum |
| UniProt Accession | Q967L9 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Multi-domain proteins (alpha and beta) |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 432 |
| Molecular Weight | 48600.5 |
| Pi | 6.17 |
| Molecular Weight | 48600.5 |
| Disulphides | 0 |
| Full Sequence |
MGSSHHHHHHSSGLVPRGSH
MAPKVQDPYA NNPSHKAFTR AFLSQSTVDF GEDNFLASPL GVLFTLGILL GSGGAQGKTS YQIAKTMRLK SANFPWNISE TQQEMKSLYK ELSDSLTMEN TTLDGNEVKV VRISTGVFTQ ETYDVESNFN KSIQNDFGGE LLTVDFTNQT NAAQDINKWV NEHSNGLVEE FFTDDIPKDA WMILINIFYF RDYWENPFVP YYTSLENFSV TNNHHLQVPM MSKDEILKYK KFPEDGFEIV SKPMKNTRFS FIILLPLEKW NLNGAIEVLN GNKILSHYVD KLEETSVSLK MPKFTLTKKL QLVGTLKSIG IKNLFDPLKA DLSGISSKNN LYVNSFIQTN VMKVNESGIE EASVTSPIIV PISALIPDVN FHVTHPFICF IYDQQLTMPL LAAKVINPML RAPPPPPLRSGC
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Yan Y, Liu S, Song G,Xu Y, Dissous C (2005) Veterinary Parasitology, 131, 53-60 |
| Project Aim | Vaccine studies |
| Fusion | N-terminal hexahistidine tag |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | None |
| Expression Temp | 16.0 |
| Expression Time | overnight |
| Expression Vector | pET28c |
| Expression Protocol | Cells were grown at 16degC overnight and expression was induced with 1mM IPTG |
| Method of Induction | IPTG |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Column refolding: Nickel-chelating chromatography |
| Wash Buffer | n/a |
| Solubilization Buffer | 8M urea, 20mM phosphate pH 7.4 |
| Refolding Buffer | not stated |
| Pre-Refolding Purification | None |
| Tag Cleaved | no |
| Refolding pH | 7.4 |
| Refolding Temperature | 4.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | Not specified |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Cells were harvested and dirsupted by sonication in buffer (20mM phosphate pH 7.4, 10mM imidazole). Proteins were dissolved completely with solubilization buffer, followed by affinity purification on a His-Trap column |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |