ANDROGEN RECEPTOR GENE IN GROWTH AND DIFFERENTIATION

仓鼠乳腺癌细胞系(DDT-1-MF-2) 含有雄激素受体，受雄激素刺激 成长。在雄激素甲基三烯烯存在的情况下， 雄激素受体水平上升至稳定状态 浓度为20fmoles/Mug DNA(2.0毫克受体/10-11 单元格)。这些细胞的雄激素受体将是 用制备性变性平板凝胶组合进行提纯 电泳层析、亲和层析、亲和层析 标记物及二维聚丙烯酰胺凝胶的制备 电泳法。噬菌体Lambdagt 11互补DNA文库 已经制备了超过25万个独立的重组人 从雄激素刺激的DDT-1中分离的信使RNA 细胞。这个库已经被证明包含完整的长度 插入到其他低丰度信使RNA中。纯洁的 雄激素受体将用于测定氨基酸 各种多肽的组成和氨基酸序列。 几个寡核苷酸探针(20-40个核苷酸长)将 密码子冗余度最低。这些探测器将是 用来筛选Lambdagt11-cDNA库，并分离和 测定雄激素受体基因的c DNA序列。CDNA3 将被用来分离基因组片段 雄激素受体基因(S)，并鉴定启动子区(S)。 这只仓鼠雄激素受体mRNA的cDNA也将是 用于筛选人源Lambdagt11-cDNA库 良性前列腺增生症信使RNA。然后，cDNA将会 用于研究雄激素受体基因在生长发育过程中的表达 在正常组织和异常组织中以及在 在各种雄激素反应系统中的细胞。

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.