Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1742
orotate phosphoribosyltransferase, pyrE

Lot Information:

Purification Lot: PC011742-41-02-02
sPlasmid ID: PC011742-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Ohmori M.
Purified Date: 08/09/2006
Total Protein Yield: 4 mg
Final Volume: 0.40 mL
Conc.: 10 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

07/22/2006Rosetta834(DE3)6 L

Cell Lysis and Heat Treatment:

20.4g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 50mM NaCl (pH8.0)
Heat Treatment at 70°C for 10 min
Ultra Cengrifuge at 200,000G for 1 hr

Purification Procedure:

ColumnBufferFlow Rate (mL/min)Eluted
Resource PHE 6ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO450.55 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.11 M NaCl
Hydroxyapatite CHT2-I10 mM NaPi (pH 7.0), 150 mM NaCl20.06 M NaPi
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.561 ml