Refolding Record:
| Protein | |
|---|---|
| Protein Name | Nucleoprotein |
| Abbreviated Name | protein N |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Hantaan virus |
| UniProt Accession | P05133 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 431 |
| Molecular Weight | 48142.3 |
| Pi | 6.63 |
| Molecular Weight | 48142.3 |
| Disulphides | Unknown |
| Full Sequence |
MATMEELQREINAHEGQLVIARQKVRDAEKQYEKDPDELNKRTLTDREGVAVSIQAKIDELKRQLADRIATGKNLGKEQDPTGVEPGDHLKERSMLSYGNVLDLNHLDIDEPTGQTADWLSIIVYLTSFVVPILLKALYMLTTRGRQTTKDNKGTRIRFKDDSSFEDVNGIRKPKHLYVSLPNAQSSMKAEEITPGRYRTAVCGLYPAQIKARQMISPVMSVIGFLALAKDWSDRIEQWLIEPCKLLPDTAAVSLLGGPATNRDYLRQRQVALGNMETKESKAIRQHAEAAGCSMIEDIE
SPSSIWVFAGAPDRCPPTCLFIAGIAELGAFFSILQDMRNTIMASKTVGTSEEKLRKKSSFYQSYLRRTQSMGIQLGQRIIVLFMVAWGKEAVDNFHLGDDMDPELRTLAQSLIDVKVKEISNQEPLKL
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Gött P, Stohwasser R, Schnitzler P, Darai G, Bautz EK. (1993) Virology, 194, 332-337 |
| Project Aim | Undefined |
| Fusion | C-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 | 0.0 |
| Expression Time | 4 h |
| Expression Vector | pQE-12 |
| Expression Protocol | After induction with 1 mM IPTG for 4 h, about 10% of total proteins of strain SG13009were found to be recombinant N proteins. |
| Method of Induction | IPTG |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | Not stated |
| Lytic Agent | None |
| Pre-Refolding Purification | Ni-NTA column |
| Solubility | partial |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | 2 M urea |
| Solubilization Buffer | 8 M urea |
| Refolding Buffer | Tris-HC1 buffer containing 4,2, and 0.3 M urea , glycerol 40% |
| Pre-Refolding Purification | Ni-NTA column |
| Tag Cleaved | no |
| Refolding pH | 0.0 |
| Refolding Temperature | 0.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The insoluble protein fractions of both construct were further enriched for the N proteins by washing the pellets with 2 M urea. N proteins solubilized in 8 M urea ere processed through a Ni-nitrilotriacetate (Ni-NTA) column, and eluted by a pH step gradient in phosphate buffer containing 8 M urea and 10 mM 2-mercaptoethanol. After a wash at pH 6.1, the majority of the N proteins was eluted at pH 5.5. Renaturation of proteins was done by stepwise dialysis against three changes of Tris-HCl buffer containing 4,2,and 0.3 M urea plus 40% glycerol |
| Refolding Assay | Unspecified |
| Refolding Chaperones | None |
| Refolding Additives | Glycerol |
| Additives Concentration | 40% |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | Refolding tem. pH. not stated |