GENE MAPPING IN THE SPONTANEOUSLY HYPERTENSIVE RAT

自发性高血压大鼠的基因图谱

• 批准号: 3736501
• 负责人: THEODORE W KURTZ
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3736501
• 关键词: animal population genetics; computer program /software; familial hypertension; genetic mapping; genetic markers; genetic polymorphism; genetic regulation; genetic strain; polymerase chain reaction; restriction fragment length polymorphism; single strand conformation polymorphism; southern blotting; spontaneous hypertensive rat

In the spontaneously hypertensive rat (SHR), it has been suggested that as few as 1-6 major genes may be determining of increased blood pressure (BP) and that the identification of these genes might shed light on the pathogenesis of essential hypertension in humans. Accordingly, we propose to: 1) develop a genetic linkage map in the rat and 2) use the map to begin searching for quantitative trait loci (QTLs) that contribute to the pathogenesis of hypertension in the SHR. Specifically, in studies in F2 populations derived from the SHR and selected inbred normotensive strains (Brown-Norway, Lewis, and Wistar-Kyoto), and in 37 recombinant inbred (RI) strains derived from the SHR and the inbred normotensive Brown-Norway rat, we will: 1) Develop a genetic linkage map in the rat by analyzing the inheritance of multiple polymorphic DNA markers. Over 40 polymorphic markers have already been developed and tested in the F2 or RI populations. Additional markers will be identified using single strand conformation polymorphism (SSCP) analysis of specific rat gene sequences, PCR analysis of microsatellite ("CA" type) repeats associated with specific rat gene sequences, and Southern blot analysis of RFLPs associated with single-locus and multi-locus tandem repeat elements marking anonymous gene sequences. The DNA markers will be localized to individual rat chromosomes by somatic-cell hybrid analysis and by linking to other markers with known chromosomal locations. Standard computer programs (MAPMAKER and LINKAGE) will be used to perform the linkage analysis and construct the map. The initial goal will be to have markers spaced at 50 Cm intervals and eventually at 20-30 Cm intervals for use in QTL likelihood mapping. 2) Obtain computerized measurements of arterial pressure in the unanesthetized, unrestrained state. 3) Search for intervals that contain genes regulating blood pressure by constructing QTL likelihood maps (using the MAPMAKER-QTL program to analyze the BP and genetic marker data). 4) Begin to isolate QTLs regulating blood pressure by creating congenic strains that are genetically identical except at the target QTL and a short length of associated chromosome. Such lines could then be used to further localize the QTLs, study their biochemical and physiological effects, and lead to the eventual cloning of specific genes determining of increased BP.

在自发性高血压大鼠（SHR）中，有人认为 只有1-6个主要基因可能决定血压升高 (BP)这些基因的鉴定可能会揭示 人类原发性高血压的发病机制。因此我们 建议：1）在大鼠中建立遗传连锁图，2）使用 开始寻找数量性状基因座（QTL）， 参与了SHR高血压的发病机制。具体而言， 在来自SHR和选择的近交系的F2群体的研究中， 正常血压株（Brown-Norway、刘易斯和Wistar-Kyoto），37例 衍生自SHR的重组近交（RI）品系和近交 正常血压的Brown-Norway大鼠，我们将：1）开发遗传连锁图谱 在大鼠中通过分析多重多态性DNA的遗传 标记。已经开发了40多个多态性标记， 在F2或RI群体中测试。其他标记将 采用单链构象多态性（SSCP）分析鉴定 特异性大鼠基因序列的PCR分析，微卫星（“CA”） 与特定大鼠基因序列相关的重复序列，以及Southern 与单位点和多位点相关的RFLPs的印迹分析 标记匿名基因序列的串联重复元件。DNA标记 将通过体细胞杂交定位到单个大鼠染色体上 分析并通过与已知染色体的其他标记相关联， 地点标准计算机程序（MAPMAKER和LINKAGE）将 用于进行连锁分析和构建图谱。初始 我们的目标是将标记以50 Cm的间隔隔开， 20-30 Cm间隔用于QTL似然作图。2)获得 在未麻醉的情况下， 不受约束的状态3)搜索包含基因的区间 通过构建QTL似然图（使用 MAPMAKER-QTL程序分析BP和遗传标记数据）。四、 开始分离调节血压的QTL， 除了在靶QTL处之外遗传上相同的菌株， 相关染色体长度短。这样的线路可以用来 进一步定位QTL，研究其生理生化特性 影响，并导致最终克隆特定的基因决定 血压升高。