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Following incubation, the resin was washed thrice on a midi batch purification column (Generon, Maidenhead, UK) using wash buffer (20 mM Tris-HCl, pH 7.8, 0.5 M NaCl, 5 M urea, 10-50 mM imidazole) before elution in buffer containing 20 mM Tris-HCl, pH 7.8, 0.5 M NaCl, 300 mM imidazole and 5 M urea

Cells were then harvested in an ice-cold hypotonic lysis buffer, passed through a 26-G needle and centrifuged at 10 000 g for 25 min at 4C

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Typically, a patient with GS is asymptomatic, without experiencing abdominal pain, pruritus, pale stools, or dark urine