Refolding Record:
| Protein | |
|---|---|
| Protein Name | Basic membrane protein D |
| Abbreviated Name | BmpD |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Borrelia burgdorferi |
| UniProt Accession | Q9FD93 |
| 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 | 374 |
| Molecular Weight | 40983.9 |
| Pi | 6.19 |
| Molecular Weight | 40983.9 |
| Disulphides | 0 |
| Full Sequence |
MLKKVYYFLIFLFIVACSSSDDGKSGAKTVSLIVDGAFDDKGFNESSSKAIRKLKADLNINIIEKASTGNSYLGDIANLEDGNSNLIWGIGFRLSDILFQRASENVSVNYAIIEGVYDEIQIPKNLLNISFRSEEVAFLAGYFASKASKTGKIGFVGGVRGKVLESFMYGYEAGAKYANSNIKVVSQYVGTFGDFGLGRSTASNMYRDGVDIIFAAAGLSGIGVIEAAKELGPDHYIIGVDQDQSYLAPNNVIVSAVKKVDSLMYSLTKKYLETGVLDGGKTMFLGLKEDGLGLVLNENLKSNYSEIYNKSLKIGQSIMNGIIKVPYDKVSYDNFVLQMENGRSNSPLAMAISDPNSSSVDKLAAALGHHHHHH
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Bryksin, A. V., Godfery, H. P., Carbonaro, C. A., Wormser, G. P., Aguero-Rosenfeld, M. E., Cabello, F. C (2005) Clin Diagn Lab Immunol, 12, 935-940 |
| Project Aim | Structure-Function |
| Fusion | C-terminal hexahistidine tag |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21-CodonPlus-RIL |
| Expression Temp | 37.0 |
| Expression Time | 2h |
| Expression Vector | pET30 Xa/LIC |
| Expression Protocol | Cells were grown in 2YT medium and induced for two hours. After lysis, inclusion bodies were collected by centrifugation and washed with washing buffer. |
| Method of Induction | IPTG |
| Cell Density at Induction | OD 600 = 0.7 |
| Cell Disruption Method | Not stated |
| Lytic Agent | None |
| Pre-Refolding Purification | Metal affinity chromatography |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | PBS, 0.05% Triton X-100 |
| Solubilization Buffer | 7M urea, 10mM DTT |
| Refolding Buffer | PBS, 0.05% Triton X-100 |
| Pre-Refolding Purification | Metal affinity chromatography |
| Tag Cleaved | no |
| Refolding pH | 7.4 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | DTT |
| Redox Agent Concentration | 10mM |
| Refolding Protocol | Inclusion bodies were washed and solubilized. rBMP proteins were purified on a HisBind Quick (Novagen) and refolded in refolding buffer dialyzed against Ficoll 400. |
| Refolding Assay | Immunoassay |
| Refolding Chaperones | None |
| Refolding Additives | Triton X-100 |
| Additives Concentration | 0.05% |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |