IDENTIFICATION OF HUMAN GLUCOCORTICOID RECEPTOR COMPLEMENTARY-DNA CLONES BY EPITOPE SELECTION
IDENTIFICATION OF HUMAN GLUCOCORTICOID RECEPTOR COMPLEMENTARY-DNA CLONES BY EPITOPE SELECTION
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DOI：
10.1126/science.2581314
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发表时间：
1985-01-01
期刊：
影响因子：
56.9
通讯作者：
EVANS, RM
中科院分区：
文献类型：
--
作者：
WEINBERGER, C;HOLLENBERG, SM;EVANS, RM
类固醇激素主要通过基因转录调节细胞分化和生理功能。通过特定类固醇受体蛋白和靶基因的相互作用来实现调节。表达克隆技术用于选择人糖皮质激素受体互补DNA克隆，以确定受体发挥其转录控制的机制。使用来自单个克隆的免疫化融合蛋白来选择表位特异性抗体，随后通过结合细胞提取物的蛋白质图来洗脱和鉴定所述表位特异性抗体。分离了三个含有表达人糖皮质激素受体抗原决定簇的插入片段的交叉杂交克隆。
Steroid hormones regulate cellular differentiation and physiologic functions predominantly through gene transcription. Regulation is achieved by the interaction of specific steroid receptor proteins and target genes. Expression cloning techniques were used to select human glucocorticoid receptor complementary DNA clones in order to define the mechanism by which the receptor exerts its transcriptional control. Immunobilized fusion proteins from individual clones were used to select epitope-specific antibody which was subsequently eluted and identified by binding to protein plots of cellular extracts. Three cross-hybridizing clones containing inserts expressing antigenic determinants of the human glucocorticoid receptor were isolated.