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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST1493
Products phosphoribosylaminoimidazole-succinocarboxamide synthase
Plasmid ID 0363
Vector  
Purification Lot ST1493
Purified Date 2006.1.25-1.31
Protein conc. (mg/ml) 23.71
Total volume (ml) 0.98
Total protein (mg) 23.2

Culture Lot:

date host cell scale label weight of bacteira (g)
        13.8

Cell Lysis and Heat Treatment:

13.8 g of E. coli.
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80ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Sonication, output 8, duty 50/3 min, 5 times.
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Heat 10 min, 76 deg C.
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Centrifuge at 100,000 rcf, 60 min.
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68 ml of supernatant.
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Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 Resource Iso 6ml 20mM Tris-HCl(pH8.0),5mM B-ME,1.35M AS 1.35M AS 0M AS 6 1.1M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 2M NaCl 4 0.7M NaCl
3 Bio-Scale CHT10I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 500mM KPi 2 from 120.4mM KPi to 239mM KPi
4 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 67min
5            
6            
7            
8            
9            

Comment:

Products was confirmed by MS.

2011.10.20

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