金属硫蛋白融合蛋白的分离纯化

PURIFICATION OF THE MOUSE BALB/C METALLOTHIONEIN-1 FUSION PROTEIN

采用IPTG诱导新 1 #菌BL2 1 (DE3) (pET2 1a MT)高效表达Balb/C小鼠金属硫蛋白融合蛋白 ,并采用包涵体分离纯化方法及SephadexG 75柱层析得到SDS

Strain new No.1〔BL21(DE3),pET21a MT,Amp〕was induced by 1 mmol/L IPTG to express the mouse Balb/C metallothionein 1 fusion protein.The method of the fusion protein purification was as following:strain new No.1 was disrupted with a sonifier and incubated at 37℃ for half an hour together with sodium deoxycholic acid,centrifugated twice.The pellet was dissolved in 8 mol/L urea denaturation solution.The inclusion body protein was renatured with the method of urea dialysis.The renatured solution was applied to a Sephadex G 75 column which had been equilibrated and eluted with the buffer 10 mmol/L Tris HCl(pH8 0) The Balb/C mouse metallothionein 1 fusion protein which showed a single band on 12% SDS PAGE after Sephadex G 75 column chromatograph was collected