Refolding Record:
| Protein | |
|---|---|
| Protein Name | Yersiniabactin transcriptional regulator |
| Abbreviated Name | YbtA |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Yersinia enterocolitica |
| UniProt Accession | Q9X9I4 |
| 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 | 339 |
| Molecular Weight | 37881.3 |
| Pi | 8.49 |
| Molecular Weight | 37881.3 |
| Disulphides | 0 |
| Full Sequence |
MTESPQTQSEISINQLVVGKPANDGNIPAQCELLRCSLQEGMDILLWRGHFARPETLQLHDDLGRINFSCILEGTSRFAIQGLRRRTDWELARNRHYITHTPDCRGSASYCGRFESITLSFSPETLALWVPDISAVIKNKIDSHCCCLQQRCNAETHLTAQALRHALMRMHGGFSHEQKPSTLWLQGQSLVMLSLVLDEHREDASCLSCHFNPMERQKLLRAKDLLLADLTQAPGVAELARESGLSVLKIKRGFRVLFNNSVYGLFQAERMQEARRRLANGNTSVMTVAADLGYANASHFSAAFQKQFGVTPSTFKRVMDPAANKARKEAELAAATAEQ
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Anisimov, R., Brem, D., Heesemann, J., Rakin, A. (2005) Int J Med Microbiol., 295, 19-28 |
| Project Aim | Functional Studies |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21(DE3)CodonPlus |
| Expression Temp | 37.0 |
| Expression Time | 6h |
| Expression Vector | pET3c |
| Expression Protocol | Cells were grown in LB medium containing 100 microg/ml ampicillin and 20 microg/ml chloramphenicol. |
| Method of Induction | IPTG |
| Cell Density at Induction | OD 600 = 0.5 |
| Cell Disruption Method | French Press |
| Lytic Agent | None |
| Pre-Refolding Purification | Washing inclusion body |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Column refolding: Size-exclusion chromatography |
| Wash Buffer | 20 mM TrisHCl, pH 8.0, 1% (v/v) Triton X-100 |
| Solubilization Buffer | 20 mM TrisHCl, pH 8.0 with 2% sodium N-lauroyl sarcosine |
| Refolding Buffer | 20 mM TrisHCl, pH 8.0 with 0.1% N-lauroyl sacrosine |
| Pre-Refolding Purification | Washing inclusion body |
| Tag Cleaved | no |
| Refolding pH | 8.0 |
| Refolding Temperature | 37.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | Beta-mercaptoethanol |
| Redox Agent Concentration | 10 mM |
| Refolding Protocol | Isolated inclusion bodies were washed twice with washing buffer, before being solubilized. The solution was then fractionated on a Superdex-200 column. |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |