Refolding Record:
| Protein | |
|---|---|
| Protein Name | Glycoprotein (recombinant glycoprotein vaccine) |
| Abbreviated Name | VHSVg |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Viral hemorrhagic septicemia virus |
| UniProt Accession | Q77AU2 |
| 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 | 508 |
| Molecular Weight | 57068.0 |
| Pi | 6.75 |
| Molecular Weight | 57068.0 |
| Disulphides | Unknown |
| Full Sequence |
MEWNTFFLVILIIIIKSTTPQITQRPPVENISTYHADWDTPLYTHPSNCRDDSFVPIRPAQLRCPHEFEDINKGLVSVPTRIIHLPLSVTSVSAVASGHYLHRVTYRVTCSTSFFGGQTIEKTILEAKLSRQEATDEASKDHEYPFFPEPSCIWMKNNVHKDITHYYKTPKTVSVDLYSRKFLNPDFIEGVCTTSPCQTHWQGVYWVGATPKAHCPTSETLEGHLFTRTHDHRVVKAIVAGHHPWGLTMACTVTFCGTEWIKTDLGDLIQVTGPGGTRKLTPNKCVNTDIQMRGATDDFS
YLNHLITNMAQRTECLDAHSDITASGKVSSFLLSKFRPSHPGPGKAHYLLDGQIMRGDCDYEAVVSINYNRAQYKTMNNTWKSWKRVDNNTDGYDGMIFGDKLIIPDIEKYQSVYDSGMLVQRNLVEVPHLSIVFVSNTSDLSTNHIHTNLIPSDWSFNWSLWPSLSGMGVVGGAFLLLVLCCCCKASPPIPNYGIPMQQFSRSQTV
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Byon JY, Ohira T, Hirono I, Aoki T. (2006) Vaccine, 24, 921-930 |
| 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 | BL21(DE3) |
| Expression Temp | 0.0 |
| Expression Time | 00 |
| Expression Vector | pET-VHSg |
| Expression Protocol | The recombinant VHSg protein was prepared by transforming Escherichia coli BL21 codon-plus competent cells with the plasmid pET-VHSg. The expression of recombinant VHSg was then induced by 1 mM isopropyl-β-thiogalactoside (IPTG). |
| Method of Induction | IPTG |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | Not stated |
| Lytic Agent | None |
| Pre-Refolding Purification | Ion-exchange chromatography |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | 8 M urea |
| Refolding Buffer | phosphate buffer saline (PBS) |
| Pre-Refolding Purification | Ion-exchange chromatography |
| 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 VHSg protein was dissolved using denaturation buffer (8 M urea), subequently purified using a His-Resin column (Qiagen, Germany) and then refolded in phosphate buffer saline (PBS) buffer. |
| Refolding Assay | Unspecified |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | the refolding temperature and pH are not stated in this paper |