CLONING AND EXPRESSION OF INFLUENZA VIRAL RNA SEGMENTS

流感病毒从顶端表面萌发的极化 上皮细胞。萌芽的机制是 仅限于根尖表面的情况仍不清楚。我们已经展示了 流感病毒包膜糖蛋白要么是血凝素 (HA)或神经氨酸酶(NA)在克隆的cDNA中表达 在没有其他病毒的情况下也会被输送到根尖表面 蛋白质。我们的目标是定义结构特征(“信号”) HA和NA参与运输、分类和极化 表情。我们将继续研究不同的功能 HA和NA的结构域，并在 插入DNA的核苷酸序列达到所需 多肽的氨基酸序列发生变化。这些 改变的cdna克隆将被表达，这些克隆的命运 改变后的蛋白质将会被追踪。因此，我们希望确定 透明质酸和氨基酸序列的特定结构域(或氨基酸序列)的作用 NA在表达、分类和运输方面以及在生物方面 这两种重要糖蛋白的功能。我们计划 继续使用真核表达载体表达这些 动物细胞中的蛋白质。我们使用的是特定部位的突变， 缺失，以及不同基因之间的嵌合融合 实现这些目标。此外，我们还表达了流感 酵母中的HA和NA病毒，它可能 在开发改进的亚基方面具有潜在的用途 疫苗。

Influenza viruses bud from the apical surface of polarized epithelial cells. The mechanism by which the budding is restricted to the apical surface remains unknown. We have shown that influenza viral envelope glycoproteins either hemagglutinin (HA) or neuraminidase (NA) when expressed from cloned cDNAs are also transported to the apical surface in absence of other viral proteins. Our goal is to define the structural features ("signals") of HA and NA involved in transport, sorting and polarized expression. We wll continue studying the function of different domains of HA and NA and make specific alterations in the nucleotide sequence of the insert DNA to achieve the desired changes in the amino acid sequence of the polypeptides. These altered cDNA clones will be expressed and the fate of these altered proteins will be followed. Thus, we hope to determine the role of the specific domains (or amino acid sequences) of HA and NA in expression, sorting, and transport as well as in the biological functions of these two important glycoproteins. We plan to continue using eukaryotic expression vectors to express these proteins in animal cells. We are using site-specific mutations, deletions, as well as chimeric fusions between different genes to achieve these objectives. In addition, we are expressing influenza viral HA and NA in yeast (Saccharomyces cerevisiae), which may be of potential use in the development of an improved subunit vaccine.