Berger F G, Gross K W, Watson G
J Biol Chem. 1981 Jul 10;256(13):7006-13.
Clones containing sequences corresponding to testosterone-inducible RNAs of mouse kidney have been identified within a cDNA clone bank prepared from size-fractionated poly(A)-containing kidney RNA. A novel screening method was employed to specifically detect such sequences. One of these, pMK908, containing a 1.2 kilobase pair insert, was studied in detail. RNA blotting experiments show that the inserted DNA of pMK908 hybridizes to two kidney-specific RNA sequences, 2500 and 1500 nucleotides in length, both of which are rapidly induced by testosterone treatment of female mice in vivo. RNA excess hybridization studies reveal that these RNAs constitute approximately 0.004% of th total RNA population in female kidneys and are induced approximately 7-8-fold by testosterone. The RNA has been isolated and translated in a cell-free system and encodes a 43,000-dalton polypeptide chain. The induction of these RNAs by testosterone is independent of the pituitary gland, but does depend upon the testosterone receptor protein. The existence of pMK908, and other chimeric plasmids containing testosterone-inducible sequences, should be valuable in understanding the response of the mouse kidney to androgens.
在从小鼠肾脏中经大小分级分离得到的含聚腺苷酸的RNA构建的cDNA克隆文库中，已鉴定出含有与小鼠肾脏睾酮诱导RNA相对应序列的克隆。采用了一种新的筛选方法来特异性检测此类序列。其中之一pMK908含有一个1.2千碱基对的插入片段，对此进行了详细研究。RNA印迹实验表明，pMK908插入的DNA与两个肾脏特异性RNA序列杂交，长度分别为2500和1500个核苷酸，在体内用睾酮处理雌性小鼠时，这两个序列均会迅速被诱导。RNA过量杂交研究表明，这些RNA约占雌性肾脏总RNA群体的0.004%，并且经睾酮诱导后增加约7 - 8倍。该RNA已被分离并在无细胞系统中进行翻译，编码一条43000道尔顿的多肽链。这些RNA受睾酮诱导独立于垂体，但确实依赖于睾酮受体蛋白。pMK908以及其他含有睾酮诱导序列的嵌合质粒的存在，对于理解小鼠肾脏对雄激素的反应应该是有价值的。
