Refolding Record:
| Protein | |
|---|---|
| Protein Name | Prolactin I |
| Abbreviated Name | PRL-I |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Oreochromis niloticus |
| UniProt Accession | P09319 |
| 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 | 215 |
| Molecular Weight | 23530.3 |
| Pi | 9.13084 |
| Molecular Weight | 23530.3 |
| Disulphides | Unknown |
| Full Sequence |
MAQRRTSGTNLFMTVLCVVAMCRAVPINELFERASQHSDKLHSLSTTLTQELDSHFPPIG
RVIMPRPAMCHTSSLQTPIDKDQALQVSESDLMSLARSLLQAWSDPLVVLSSSASTLPHP
AQSTIFNKIQEMQQYSKSLKDGLDVLSSKMGSPAQAITSLPYRGGTNLGHDKITKLINFN
FLLSCLRRDSHKIDSFLKVLRCRAAKMQPEMC
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Swennen D, Rentier-Delrue F, Auperin B, Prunet P, Flik G, Wendelaar Bonga SE, Lion M, Martial JA. (1991) J Endocrinol, 131, 219-227 |
| 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 |
| Expression Temp | 37.0 |
| Expression Time | 4h |
| Expression Vector | pARAE |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | French Press |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | 10mM Tris-HCl, 1mM EDTA, 0.1mM PMSF pH 8.0 |
| Solubilization Buffer | 20mM NH4HCO3, 6M urea, 1% betamercaptoethanol pH 10 |
| Refolding Buffer | 20mM NH4HCO3 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 10.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | Beta-mercaptoethanol |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Inclusion bodies were washed three times in 500mls of 10mM Tris-HCl, 1mM EDTA, 0.1mM PMSF pH 8.0. The protein was solubilised in 20mM NH4HCO3, 6M urea, 1% betamercaptoethanol pH 10 and refolded by dialysis against 100 vol. of 20mM NH4HCO3 at a flow rate of 2L per hour. |
| Refolding Assay | Ligand Binding |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 10-15mg/L culture |
| Purity | |
| Notes | |