摘要
The homogeneity between the high-and low-affinity glucocorticoid receptors(GR_H and GR_L)was testified by immunoaffinity chromatography using Mab N250(recognizing the immunogenic domain at the N terminal of GR_H)and Mab BuGR1(recognizing the DNA binding domain of GR_H).The specific binding peak of 0.98μmol/L[~3H]triamcinolone acetonide(TA)was higher than that of 52.50nmol/L[~3H]TA.The re-sult suggests that the antigenic determinants of GR_L are similar to those of GR_H.ThecDNA of rat liver GR_H was introduced into GR_H-and GR_L-negative mouse fibroblast cellline E82.A3 by calcium phosphate coprecipitation.A number of clones which expressGR_H were selected with G418(400μg/ml).The results of radioligand binding assay indicatethat GR_H gene is expressed successfully and GR_L also may be encoded by GR_H gene.
The homogeneity between the high-and low-affinity glucocorticoid receptors (GR_H and GR_L)was testified by immunoaffinity chromatography using Mab N250 (recognizing the immunogenic domain at the N terminal of GR_H)and Mab BuGR1 (recognizing the DNA binding domain of GR_H).The specific binding peak of 0.98μmol/L [~3H]triamcinolone acetonide(TA)was higher than that of 52.50nmol/L[~3H]TA.The re- sult suggests that the antigenic determinants of GR_L are similar to those of GR_H.The cDNA of rat liver GR_H was introduced into GR_H-and GR_L-negative mouse fibroblast cell line E82.A3 by calcium phosphate coprecipitation.A number of clones which express GR_H were selected with G418(400μg/ml).The results of radioligand binding assay indicate that GR_H gene is expressed successfully and GR_L also may be encoded by GR_H gene.
