建立全长人PPARγ重组蛋白的固相金属亲和层析纯化方法。利用E.coli BL_(21)(DE3)细胞外源性表达出全长人PPARγ重组蛋白,采用固相金属亲和层析法纯化目标蛋白。通过考察咪唑浓度对纯化的影响,确定在8 mol/L尿素下,用含50 mmol/L咪唑的缓冲液冲洗,200mmol/L咪唑缓冲液洗脱,可获得纯度为95%的目标蛋白,透析复性后经放射配体受体饱和结合实验测定全长人PPARγ重组蛋白的解离常数K_d为106±6nmol/L。结果表明本文所建立的方法能够成功纯化出高纯度的目标蛋白,经复性后,可获得用于结构和功能研究的全长人PPARγ重组蛋白。 To establish a full-length human PPARγ recombinant protein by solid-phase metal affinity chromatography purification method. The full length human PPARγ recombinant protein was expressed exogenously in E. coli BL_ (21) (DE3), and the target protein was purified by solid phase metal affinity chromatography. By investigating the effect of imidazole concentration on purification, it was confirmed that the target protein with a purity of 95% could be obtained under the condition of 8 mol / L urea, washing with 50 mmol / L imidazole buffer and 200 mmol / L imidazole buffer, After refolding, the dissociation constant of the full-length human PPARγ recombinant protein K_d was 106 ± 6 nmol / L as determined by the binding of radioligand receptors to saturation. The results show that the method established in this paper can successfully purify the target protein with high purity, and after annealing, the full-length human PPARγ recombinant protein can be obtained for structural and functional studies.