CLONING AND EXPRESSION OF GAP JUNCTION CHANNELS

缝隙连接介导的细胞内通讯被认为是潜在的 一些关键的细胞行为。这些包括电子产品的传播 和细胞群体之间的激素信号和生长和调节 发展。DNA克隆表明存在一些相关的差距 连接蛋白，我们通常称之为连接蛋白。1)功能界别 许多已鉴定的连接蛋白形成的缝隙连接的性质 还没有被描述。我们将使用体外表达系统 由一对电压钳制的非洲爪哇卵母细胞组成的研究 属性。杂交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)GAP的同一性 晶状体中的连接蛋白仍不清楚。有报道称， 26kD蛋白(MP26)和70kD蛋白(MP70)是缝隙连接 结构蛋白。此外，我们还克隆了CDNA 晶状体中的另一种缝隙连接蛋白。我们将制造这种新的抗体 蛋白质，并确定其分布。我们将克隆MP70的cDNA以 确定其与MP26和其他可能的缝隙连接蛋白的关系 3)我们发现了一种缝隙连接蛋白，它的mRNA只存在于 非洲爪哇早期胚胎。我们将确定空间和时间 该蛋白的表达及其在早期表达的尝试 非洲爪哇的胚胎。4)我们将克隆缝隙连接蛋白的编码基因 在果蝇身上。可用于操纵的强大技术 在这种有机体中的基因表达将允许仔细检查可能的 缝隙连接交流在生长发育中的作用。

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.