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| Refolding Record [Created: 2009-01-21 Updated: 2009-01-23 ] |
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PROTEIN |
| Protein Name |
non-POU domain-containing octamer binding protein |
| Abbreviated Name |
NonO |
| SCOP Family |
Unknown |
| Structure Notes |
MTMITPSLKDHLIHNVHKEEHAHAHNKIDDDDKVNQSNKTFNLEKQNHTPRKHHQHHHQQHHQQQQQQQQQQQQQPPPPIPANGQQASSQNEGLTIDLKNFRKPGEKTFTQRSRLFVGNLPPDITEEEMRKLFEKYGKAGEVFIHKDKGFGFIRLETRTLAEIAKVELDNMPLRGKQLRVRFACHSASLTVRNLPQYVSNELLEEAFSVFGQVERAVVIVDDRGRPSGKGIVEFSGKPAARKALDRCSEGSFLLTTFPRPVTVEPMDQLDDEEGLPEKLVIKNQQFHKEREQPPRFAQPGSFEYEYAMRWKALIEMEKQQQDQVDRNIKEAREKLEMEMEAARHEHQVMLMRQDLMRRQEELRRMEELHNQEVQKRKQLELRQEEERRRREEEMRRQQEEMMRRQQEGFKGTFPDAREQEIRMGQMAMGGAMGINNRGAMPPAPVPPGTPAPPGPAAMMPDGTLGLTPPTTERFGQAATMEGIGAIGGTPPAFNRPAPGAEFAPNKRRRY |
| Organism |
Rat |
| UniProt Accession |
Q5FVM4 |
| SCOP Unique ID |
n/a |
| Structure Solved |
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| Class |
Unknown |
| Molecularity |
Unknown |
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CONSTRUCT |
| Full Length |
y |
| Domain |
n/a |
| Chimera |
n/a |
| Variants |
n/a |
| Chain Length |
510 |
| Molecular Weight |
58860.6 |
| pI |
8.33 |
| Disulphides |
Unknown |
Pfam Domain(s) and
Number of Occurrences |
Pfam Entry
NOPS(1), RRM_1(2) |
| Domain Graphics |
 |
| Full Sequence |
MTMITPSLKDHLIHNVHKEEHAHAHNKIDDDDKVNQSNKTFNLEKQNHTPRKHHQHHHQQHHQQQQQQQQQQQQQPPPPIPANGQQASSQNEGLTIDLKN
FRKPGEKTFTQRSRLFVGNLPPDITEEEMRKLFEKYGKAGEVFIHKDKGFGFIRLETRTLAEIAKVELDNMPLRGKQLRVRFACHSASLTVRNLPQYVSN
ELLEEAFSVFGQVERAVVIVDDRGRPSGKGIVEFSGKPAARKALDRCSEGSFLLTTFPRPVTVEPMDQLDDEEGLPEKLVIKNQQFHKEREQPPRFAQPG
SFEYEYAMRWKALIEMEKQQQDQVDRNIKEAREKLEMEMEAARHEHQVMLMRQDLMRRQEELRRMEELHNQEVQKRKQLELRQEEERRRREEEMRRQQEE
MMRRQQEGFKGTFPDAREQEIRMGQMAMGGAMGINNRGAMPPAPVPPGTPAPPGPAAMMPDGTLGLTPPTTERFGQAATMEGIGAIGGTPPAFNRPAPGA
EFAPNKRRRY
BLAST against Refold database | BLAST against NCBI NR database
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| Notes |
n/a |
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EXPRESSION |
| Report |
Unpublished ( 0) J Virol Methods, 0, 0 |
| Project Aim |
Protein-Protein Interaction Identification |
| Fusion |
N-terminal hexahistidine tag |
Protein Expression and
Production |
Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host |
E coli |
| Expression Strain |
BL21 |
| Expression Temperature |
37 |
| Expression Time |
2h - 12h |
| Expression Vector |
pHAT |
| Expression Protocol |
Ni-resin affinity |
| Method of Induction |
IPTG |
| Cell Density (at induction) |
OD 0.7 = 600 |
| Cell Disruption Method |
Sonication |
| Lytic Agent |
None |
| Pre-refolding Purification |
Affinity chromotography |
| Solubility |
soluble |
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REFOLDING |
| Refolding Method |
Dialysis |
| Wash Buffer |
8M Urea, 100mM potassium phosphate buffer,pH7.2 |
| Solubilization Buffer |
8M Urea, 100mM potassium phosphate buffer,pH7.2 |
| Refolding Buffer |
150mM NaCl, 50mM potassium phosphate buffer,pH8.0, 10%glycerol, 0.05% Tween 20, 2mM 2-mercaptpethanol |
| Pre-refolding Purification |
Affinity chromotography |
| Tag Cleaved |
no |
| Refolding pH |
8 |
| Refolding Temperature |
4 |
| Protein Concentration |
1 mg/ml |
| Refolding Time |
4h |
| Redox Agent |
Beta-mercaptoethanol |
| Redox Agent Concentration |
n/a, n/a |
| Refolding Protocol |
dialysis |
| Assay |
Electrophoretic mobility-shift assays |
| Chaperones |
None |
| Additives |
tween 20 |
| Additives Concentration |
n/a |
| Refolding Yield |
n/a |
| Purity |
n/a |
| Notes |
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