STRUCTURE OF THE RAT CALMODULIN GENE

大鼠钙调蛋白基因的结构

• 批准号: 3428556
• 负责人: FELIX FRIEDBERG
• 金额: $ 2.63万
• 依托单位: HOWARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-02-01 至 1988-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3428556
• 关键词: aging; amino group; brain metabolism; complementary DNA; gene expression; genetic library; genetic mapping; lymphocyte; molecular cloning; molecular pathology; neurotransmitter metabolism; nucleic acid sequence

The calcium binding protein calmodulin regulates many enzymatic processes in the brain including those that synthesize and release neurotransmitters. 1. It is proposed to study this molecule at the level of the gene in order to elucidate tht structure of the calmodulin gene in the mammalian organism. The nucleotide sequences in the protein coding region, the intron-exon junctions, and possibly in the flanking region of the gene will be determined. cDNA probes derived from a brain lambda gt 11 cDNA library will be used to screen for calmodulin clones contained in a rat genomic library. The structure and organization of the rat calmodulin gene will be determined by restriction mapping and by sequencing the DNA. 2. In collaboration with Dr. Detera-Wadleigh at the Clinical Neurogenetics Branch, MHIRP, NIMH, the isolated DNA will be used as a probe to screen for restriction fragment length polymorphisms at the calmodulin locus to determine whether such polymorphisms correlate with affective disorders and schizophrenia. Human genomic DNA is isolated from lymphoblasts. Screening is accomplished by Southern blot analysis. 3. In the hippocampal neurons of aged rats the calcium dependent after-hyperpolarizations are prolonged. Calmodulin levels in the brain decrease with age. It is not known whether calmodulin gene expression in one area of the brain is influenced by developmental and aging processes to the same extent as in other brain structures. The expression of the calmodulin gene in different parts of the rat brain at various stages of development and senescence will be investigated. Here the techique of "Northern" blotting is employed.

钙结合蛋白钙调素调节许多酶 包括那些合成和释放 神经传递素 1. 建议在基因水平上研究这种分子， 为了阐明钙调素基因的结构， 哺乳动物有机体。 蛋白质中的核苷酸序列 编码区，内含子-外显子连接，可能在 将确定基因的侧翼区。 cDNA探针 来自脑λ gt 11 cDNA文库的DNA将用于 筛选包含在大鼠基因组文库中的钙调蛋白克隆。 大鼠钙调素基因的结构和组织将是 通过限制性酶切图谱和DNA测序确定。 2. 在与Detera-Wadleigh博士的临床合作中， 神经遗传学分支，MHIRP，NIMH，分离的DNA将被 用作探针以筛选限制性片段长度 在钙调蛋白基因座的多态性，以确定是否这样的 多态性与情感障碍相关， 精神分裂症 人类基因组DNA分离自 淋巴母细胞 通过Southern印迹完成筛选 分析. 3. 在老年大鼠海马神经元中， 后超极化被延长。 心脏中的钙调素水平 大脑随着年龄的增长而减少。 目前尚不清楚钙调素基因是否 大脑某个区域的表达受到 发展和衰老过程的程度与其他 大脑结构 钙调素基因在大肠杆菌中的表达 在不同发育阶段的大鼠大脑的不同部分 并研究衰老。 这里的技术 采用“北方”印迹法。