CLONING & EXPRESSION OF GAP JUNCTION CHANNELS

克隆

• 批准号: 3293409
• 负责人: DAVID L PAUL
• 金额: $ 20.56万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-12-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3293409
• 关键词: Drosophilidae; Xenopus oocyte; antisense nucleic acid; complementary DNA; gap junctions; gel electrophoresis; gene expression; genetic library; growth /development; heart cell; immunofluorescence technique; in situ hybridization; laboratory rat; lens proteins; membrane channels; messenger RNA; molecular cloning; myocardium; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; polymerase chain reaction; protein biosynthesis; protein structure; protein structure function

Intracellular communication mediated by gap junctions is thought to underli a number of crucial cell behaviors. These include the spread of electrical and hormonal signals among populations of cells and regulation of growth an development. DNA cloning indicates that there are a number of related gap junction proteins, which we generically call connexins. 1) The functional properties of gap junctions formed from many of the identified connexins have not been described. We will use an in vitro expression system consisting of pairs of voltage clamped Xenopus oocytes to study those properties. Hybrid proteins will be functionally expressed to identify protein domains that control different behavior. 2) The identity of gap junction proteins in the lens remains unclear. There are reports suggestin that a 26 kD protein (MP26) and a 70 kD protein (MP70 are gap junction structural proteins. In addition, we have cloned CDNA for what may be another gap junction protein in lens. We will make antibodies to this new protein and determine its distribution. We will clone cDNA for MP70 to determine its relationship to MP26 and other possible gap junction proteins 3) We have identified a gap junction protein whose mRNA is present only in Xenopus early embryos. We will determine the spatial and temporal expression of the protein and attempt to modulate its expression in early Xenopus embryos. 4) We will clone cDNAs coding for gap junction proteins in Drosophila. The powerful techniques available for the manipulation of gene expression in this organism will permit close examination of possible roles of gap junctional communication in growth and development.

缝隙连接介导的细胞内通信被认为是基础的 许多关键的细胞行为。 这些包括电气的传播 细胞种群中的激素信号和生长的调节a 发展。 DNA克隆表明存在许多相关差距 连接蛋白，我们通常称其为连接蛋白。 1）功能 由许多确定的连接素形成的间隙连接的特性 尚未描述。 我们将使用体外表达系统 由一对夹紧的爪蟾卵母细胞组成的研究 特性。 混合蛋白将在功能上表达以识别 控制不同行为的蛋白质结构域。 2）差距的身份 镜头中的连接蛋白尚不清楚。 有报道暗示 26 kD蛋白（MP26）和70 kD蛋白（MP70是间隙连接 结构蛋白。 此外，我们还有克隆的cDNA 镜头中的另一种间隙连接蛋白。 我们将对这个新的抗体制作抗体 蛋白质并确定其分布。 我们将克隆cDNA的mp70至 确定其与MP26和其他可能的间隙连接蛋白的关系 3）我们已经确定了一个间隙连接蛋白，其mRNA仅存在于 爪蟾早期胚胎。 我们将确定空间和时间 蛋白质的表达并试图调节其早期表达 爪蟾胚胎。 4）我们将克隆cDNA编码为间隙连接蛋白 在果蝇中。 可用于操纵的强大技术 该生物中的基因表达将允许对可能的可能 差距连接在增长和发展中的作用。