TRANSCRIPTIONAL REGULATION OF HUMAN 17-HYDROXYLASE GENE

根据医师科学家奖(PSA)，候选人和 赞助商，与妇产科和 加州大学旧金山分校的内分泌学研究生课程构建了一个详细的 为期5年的2阶段培训计划，旨在为应聘者提供 有丰富的研究经验。该计划是HER不可分割的一部分 内分泌学博士候选人资格。第一阶段涉及完全注册 报名参加6个系的8个研究生部课程，正规 参加各种研讨会和三次实验室轮换。这些都会完成的 与应聘者作为生殖研究员的培训同时进行 内分泌科。产科和妇科。这段经历 将满足PSA的第一阶段要求，并履行 进入博士课程的候选人资格要求。第二阶段是 在主要赞助商的直接监督下进行100%的实验室研究。 这项研究也将构成申请者的博士论文。在.期间 第二阶段，候选人的研究工作将集中在转录 人类P450c17基因的调控，特异的顺式相互作用和相互作用。 导致组织特异性目标的要素和反式作用因素： (1)研究基因内序列的潜在调控作用 稳定转染法对激素调控转录的影响。到目前为止 只有5‘侧翼区域对激素反应进行了研究。 然而，在其他一些系统中，基因内序列已经被证明是 调节转录。(2)定义涉及的特定顺式元素 组织特异性和激素反应性。瞬时转染法 实验将用含有该基因片段的质粒进行。 5‘侧翼区与一个异源启动子和CAT报告基因连锁。 (3)建立内源性表达P450c17的小鼠细胞系。 对P450c17基因的研究一直受到缺乏具有 内源性P450c17的表达。我们将构建转基因小鼠， P450c17靶向表达SV40T抗原。这一方法已经被使用 通过该机构的合作者的成功建立 分化的细胞系。(4)重新定义组织中涉及的顺式元件- 瞬时细胞转染法的特异性和激素反应性 在目标3.特定核因素之间的相互作用中建立的线 并将对这些顺式元素进行调查。成功完成 这些目标将大大增加我们对荷尔蒙的理解 调节类固醇合成中这一关键酶的组织特异性表达。 成功完成这一培训计划将使应聘者具备 开创研究分子的独立研究生涯 人类繁衍的基础。

In accordance with the Physician Scientist Award (PSA), the candidate and the sponsor, in conjunction with the Dept of Obstetrics and Gynecology and the Graduate Program in Endocrinology at UCSF, have constructed a detailed 2-phase 5-year training program intended to provide the candidate with an intensive research experience. This program is an integral part of her candidacy for a Ph.D. in Endocrinology. Phase I involves fully registered enrollment in eight graduate division courses in 6 departments, regular attendance at various seminars and three lab rotations. These will be done concurrently with the candidate's training as a fellow in Reproductive Endocrinology in the Dept. of Obstetrics and Gynecology. This experience will fulfill the Phase I requirements of the PSA and fulfill the requirements to advance to candidacy in the Ph.D. program. Phase II is 100% lab research under the direct supervision of the primary sponsor. This research will also constitute the applicants Ph.D. thesis. During phase II, the candidate's research effort will focus on the transcriptional regulation of the human P450c17 gene, the interaction between specific cis- elements and trans-acting factors which leads to the tissue-specific aims: (1) Investigate the potential regulatory role of the intragenic sequences on hormonally regulated transcription by stable transfection. Thus far only the 5' flanking region has been studied for hormone-responsiveness. However, in some other systems, intragenic sequences have been shown to modulate transcription. (2) Define the specific cis-elements involved in tissue-specificity and hormone-responsiveness. Transient transfection experiments will be performed with plasmids containing fragments of the 5'flanking region linked to a heterologous promotor and CAT reporter gene. (3) Establish a murine cell line which expresses P450c17 endogenously. Study of the P450c17 gene has been limited by the lack of a cell line with endogenous P450c17 expression. We will construct transgenic mice with P450c17 targeted SV40 T-antigen expression. This approach has been used with success by collaborators at this institution to establish differentiated cell lines. (4) Redefine cis-elements involved in tissue- specificity and hormone-responsiveness by transient transfection with cell lines established in aim #3. Interaction between specific nuclear factors and these cis-elements will be investigated. Successful completion of these aims will add substantially to our understanding of the hormonally regulated tissue-specific expression of this key enzyme in steroidogenesis. Successful completion of this training program will equip the candidate to initiate an independent research career studying the molecular underpinnings of human reproduction.