【摘要】 目的　确立人甲状腺激素受体相互作用蛋白 15 (hTRIP15 )与甲状腺激素受体 (TR)的相互作用及其作用方式。方法　应用酵母双杂交技术及半乳糖苷酶定性或定量测定。结果　将构建质粒 (BD TRIP15与AD TR)共转化AH10 9酵母细胞后 ,在高严格度选择的培养基 (SD Leu Trp Ade His) 10天克隆长出 ,相比阳性对照质粒 (BD P5 3 +AD Tag)共转后 3天克隆生长。将构建载体及对照载体转化Y187酵母菌株 ,检测LacZ报告基因的表达 (β gal单位 ) ,阳性对照质粒 (BD P5 3 +AD Tag)为 14 9.5 7± 0 .5 1,BD TRIP15 +AD TR为 3 .2 3± 10 .15 ,加入T3 后为 2 .0 2± 0 .0 8,而阴性对照质粒 (BD P5 3 +AD LamC)仅为 0 .0 2± 0 .0 1β gal单位 ,且T3 呈剂量依赖方式抑制两者的相互作用。结论　hTPIR15能与TR共相互作用 ,T3 呈剂量依赖方式抑制两者间的作用 ,仅在超大剂量时才可完全阻断hTRIP15与TRα间的作用。更多还原

【Abstract】 Objective To indentify interaction of human thyroid hormone receptor interacting protein 15 (hTRIP15) and thyroid hormone receptor (TR) and their interacting manner. Methods Yeast two-hybrid technique and determination of β-galactosidase (β-gal) activity were applied. Results After two constructs, plasmids (BD-TRIP15 and AD-TR) were co-transfected into AH109 yeast strain, it took 10 days that clones appeared on high stringent-selective medium (SD-Leu-Trp-Ade-His), and by comparison, clones might grow in 3 days for co-transfection of positive control plasmids (BD-P53+AD-Tag). To determine expression (β-gal unit) of LazZ report gene, above constructs and positive control vectors were co-transfected into Y187 yeast strain. The activities of LacZ in BD-P53+AD-Tag (positive control) and BD-TRIP15+AD-TR were 149.57±0.51 and 3.23±10.15 respectively. After adding thyroid hormone T 3, BD-TRIP15+AD-TR was 2.02±0.08, and BD-P53+AD-LamC (negative control) only was 0.02±0.01 β-gal units. T 3 inhibited the interaction of hTRIP15 and TR in dosage-dependent pattern. Conclusion hTRIP15 can interact with TR, T 3 may inhibit the interaction between hTRIP15 and TR, and may block the interaction only in exorbitant dosage.更多还原