MOLECULAR GENETICS OF INFLUENZA VIRUS HAEMAGGLUTININ

这项研究的总体目的是了解它的结构和 真核细胞膜蛋白的功能分析 这类分子的各个区域的分子水平。在 去年，已经开发出克隆、插入这些基因的方法 转化为表达载体，将它们重新引入哺乳动物细胞，并 获得细胞表面真核蛋白的表达。在……里面 原则上，这些技术可以应用于任何编码一种 从正常或恶性细胞中分离出来的细胞表面蛋白。在中国的工作 这项建议涉及一种特殊的膜蛋白--一种病毒编码的 血凝素。具体目标是：1.在内部引入突变 疏水结构域、抗原区和糖基化位点 流感病毒血凝素。2.构建嵌合基因 流感血凝素的疏水区被换成 真核生物和细菌来源的其他膜蛋白。3.至 分离大量突变型和嵌合型蛋白及其活性 野生型同源物。这些蛋白质的各种生物学功能 将被确定并与它们的结构相关联，这将是 如有必要，可通过X射线结晶学进行详细分析。4.建设 含有信号和锚定疏水区的哺乳动物载体 流感血凝素。这些载体将用于在细胞上表达 通常只在内部发现的表面真核蛋白 牢房。通过这些和其他实验，人们希望找出：如何 这样的蛋白质与细胞膜相互作用；这些分子如何 糖基化以及这种糖基化可能起到什么作用；如何 膜分子被定向到细胞的特定表面，一旦 在那里，它们如何执行特定的生物功能并充当 抗原和免疫原。

The general aim of this research is to learn about the structure and functions of the membrane proteins of eukaryotic cells through analysis at the molecular level of the individual domains of such molecules. In the last year, methods have been developed to clone such genes, to insert them into expressing vectors, to reintroduce them into mammalian cells and to obtain expression of the authentic protein on the cell surface. In principle, these techniques can be applied to any gene coding for a cell-surface protein isolated from normal or malignant cells. The work in this proposal deals with one particular membrane protein - a virally-coded hemagglutinin. The specific aims are: 1. to introduce mutations within the hydrophobic domains, the antigenic regions and the glycosylation sites of influenza virus hemagglutinin. 2. to construct chimeric genes in which the hydrophobic regions of influenza hemagglutinin are exchanged for those of other membrane proteins of both eukaryotic and bacterial origin. 3. to isolate large quantities of mutant and chimeric proteins and their wild-type homologs. The various biological functions of these proteins will be determined and correlated with their structures, which will be analyzed in detail if necessary by X-ray crystallography. 4. to construct mammalian vectors containing the signal and anchor hydrophobic regions of influenza hemagglutinin. These vectors will be used to express on the cell surface eukaryotic proteins that are normally found only in the interior of the cell. By these and other experiments, it is hoped to find out: how such proteins interact with the membranes of the cell; how such molecules become glycosylated and what function such glycosylation may serve; how membrane molecules are directed to particular surfaces of the cell and once there, how they carry out specific biological functions and serve as antigens and immunogens.