GENE STRUCTURE AND FUNCTION IN THE RAT SEMINAL VESICLE

精囊分泌蛋白(SVS)IV和V在大鼠体内的表达 在雄激素的控制下。SVS IV基因和SVS基因 V是从一个大鼠基因文库中分离出来的。DNA序列已经被 确定了SVS IV的转录单位和侧翼序列 吉恩。第一个目标是推导出侧翼的DNA序列 SVS V基因的区域和转录单位。其目的是比较 SVS IV和SVS V中潜在调控位点的5‘侧翼区域 可能在雄激素作用中起一定作用的基因。 超螺旋载体pPSVS 3.3中两个主要的S1核酸酶敏感位点 (包含SVS IV的侧翼和转录单位)已被 已确认身份。5‘-Sl位点位于或接近位于-113到 -转录起始点的117个碱基。与排序一起对 SVS V基因、S1核酸酶敏感部位也将在超螺旋中定位 含有SVS V基因的质粒，因为它们被分离、定位和 已排序。 SVS IV和V中的DNase I超敏和S1核酸酶敏感部位 基因将通过消化正常和去势的细胞核来定位。 精囊组织。这些数据将与由 含SVS IV的各种超螺旋载体的S1核酸酶酶切 和SVS V基因片段。 一个主要目标是将SVS IV和SVS V基因与几个 带有显性选择标记的真核克隆载体(如PSV-neo) 并用Well将SVS基因导入受体细胞系 已鉴定的雄激素受体或转入精液原代培养 囊泡细胞。 将从转基因实验中建立细胞系， 当雄激素被加入到培养液中时， 引入的SVS基因将显著增加。用这些细胞系 以及各种体外缺失突变体的产生和转染人 这些突变的DNA形式的SVS基因进入细胞系，在 5‘-侧翼、转录单位和/或3’-侧翼 可能会建立雄激素依赖的表达。这些数据将扩大 我们关于类固醇激素作用和血管紧张素转换酶的控制 显然是协同调控的基因集合。

Seminal vesicle secretory proteins (SVS) IV and V expression in the rat are under the control of androgens. The gene for SVS IV and the genes for SVS V have been isolated from a rat gene library. The DNA sequence has been determined for the transcription unti and flanking sequences of the SVS IV gene. The first objective is to derive the DNA sequence of the flanking regions and transcription unti for the SVS V genes. The aim is to compare 5'-flanking regions for potential regulatory sites in SVS IV and SVS V genes which may have some role in androgen action. Two major Sl nuclease sensitive sites in supercoiled plasmid pSVS 3.3 (containing flanking and transcription unit for SVS IV) have been identified. The 5'-Sl site is at or near an inverted repeat at -113 to -117 bp from the transcription initiation site. Along with sequencing the SVS V genes, Sl nuclease sensitive sites will also be mapped in supercoiled plasmids containing the SVS V genes as they are isolated, mapped, and sequenced. DNAse I hypersensitive and Sl nuclease sensitive sites in the SVS IV and V genes will be mapped by digestion of nuclei from normal and castrate seminal vesicle tissue. These data will be compared to sites identified by the Sl nuclease digestion of various supercoiled plasmids containing SVS IV and SVS V gene fragments. A major objective will be to link the SVS IV and SVS V genes to several eukaryotic cloning vectors with dominant selection markers (e.g., pSV-neo) and introduce the SVS genes into recipient cell lines with well characterized androgen receptors or into primary cultures of seminal vesicle cells. Cell lines will be established from the transfection experiments which, when androgen is added to the media, the transcription rate of the introduced SVS genes will significantly increase. With these cell lines and the production of various in vitro deletion mutants and transfection of these mutant DNA forms of the SVS genes into cell lines, the regions in the 5'-flanking, transcription unit and/or 3'-flanking that are necessary for androgen-dependent expression may be established. These data will broaden our knowledge base about steroid hormone action and the control of an apparently coordinately regulated gene set.