CLONING AND EXPRESSION OF INFLUENZA VIRAL RNA SEGMENTS

流感病毒从两极分化的 上皮细胞 萌芽的机制是 仅限于根尖表面的情况仍不清楚。 我们已经表明 流感病毒包膜糖蛋白或者血凝素 (HA)或神经氨酸酶（NA），当从克隆的cDNA表达时 在没有其他病毒的情况下也被转运到顶端表面， proteins. 我们的目标是定义结构特征（“信号”） HA和NA参与运输、分选和极化 表情 我们将继续研究不同的功能 结构域的HA和NA，并作出具体的改变， 插入DNA的核苷酸序列，以实现所需的功能。 多肽的氨基酸序列的变化。 这些 改变的cDNA克隆将被表达，并且这些克隆的命运 将跟踪改变的蛋白质。 因此，我们希望确定 HA的特定结构域（或氨基酸序列）的作用， NA在表达、分选和转运以及生物学中的作用 这两个重要的糖蛋白的功能。 我们计划 继续使用真核表达载体来表达这些 动物细胞中的蛋白质 我们使用的是定点突变， 缺失，以及不同基因之间的嵌合融合， 实现这些目标。 此外，我们还表示， 酵母（Saccharomycescerevisiae）中的病毒HA和NA，其可以 在开发改进的亚单位中具有潜在的用途 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.