ANDROGEN RECEPTOR GENE IN GROWTH AND DIFFERENTIATION

仓鼠乳管癌细胞系DDT-1-MF-2 含有雄激素受体，受雄激素刺激， 成长 在雄激素甲基三烯醇的存在下， 雄激素受体水平升高至稳定状态 浓度为20 fmol/μ g DNA（2.0 mg受体/10-11 细胞）。 这些细胞中的雄激素受体 通过制备变性Slab凝胶 电泳，亲和层析，光亲和 标记和制备二维聚丙烯酰胺凝胶 电泳 噬菌体Lambdagt 11互补DNA文库 已经制备了超过250，000个独立的重组体， 从雄激素刺激的DDT-1中分离的信使RNA 细胞 该库已被证明包含全长 插入到其他低丰度信使RNA中。 纯 雄激素受体将用于测定氨基酸 肽的组成和各种肽的氨基酸序列。 将使用几种寡核苷酸探针（20-40个核苷酸长）。 以最小的密码子冗余制备。 这些探测器将 用于筛选Lambdagt 11-cDNA文库并分离和 对雄激素受体mRNA的cDNA进行测序。 的cDNA 然后将用于分离基因组片段， 雄激素受体基因并鉴定启动子区。 这种仓鼠雄激素受体mRNA的cDNA也将被克隆。 用于筛选人Lambdagt 11-cDNA文库 良性前列腺增生信使RNA。 然后cDNA将 用于研究生长过程中雄激素受体基因的表达 正常和异常组织的分化， 细胞在各种雄激素反应系统。

The hamster ductus deferns smooth tumor cell line (DDT-1-MF-2) contains androgen receptors and is stimulated by androgens to grow. In the presence of the androgen methyltrienolene, the level of androgen receptor increases to a steady state concentration of 20 fmoles/Mug DNA (2.0 mg of receptor/10-11 cells). The androgen receptor from these cells will be purified by a combination of preparative denaturing Slab gel electrophoresis, affinity chromatography, photoaffinity labelling, and preparative two-dimensional polyacrylamide gel electrophoresis. A phage Lambdagt 11 complementary DNA library with over 250,000 independent recombinant has been prepared from messenger RNA, isolated from androgen stimulated DDT-1 cells. This library has been shown to contain full length inserts to other low abundance messenger RNAs. The pure androgen receptor will be used to determine amino acid composition and the amino acid sequence of various peptides. Several oligonucleotide probes (20-40 nucleotides long) will be prepared with minimal codon redundancy. These probes will be used to screen the Lambdagt 11-cDNA library and to isolate and sequence the cDNa for the androgen receptor mRNA. The cDNA will then be used to isolate the genomic fragments for the androgen receptor gene(s) and identify the promoter region(s). The cDNA for this hamster androgen receptor mRNA will also be utilized to screen a Lambdagt 11-cDNA library made to human benign prostate hyperplasia messenger RNA. The cDNAs will then be used to study androgen receptor gene expression in growth and differentiation in both normal and abnormal tissues and in cells in a variety of androgen responsive systems.