CLONING AND EXPRESSION OF HUMAN PARAINFLUENZA VIRUS

人副流感病毒的克隆和表达

• 批准号: 2294791
• 负责人: YUMIKO MATSUOKA
• 金额: $ 24.98万
• 依托单位: SECRETECH, INC.
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-06-28 至 1992-06-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2294791
• 关键词: CLONING; EXPRESSION; HUMAN; PARAINFLUENZA; VIRUS

We have constructed cDNA libraries of human parainfluenza type 1 (PI1) and type 2 (PI2) viral mRNAs in a plasmid vector. Two glycoprotein gene clones from the cDNA libraries, HN of PI1 and F of PI2 virus have already been characterized. Screening continues for the other glycoprotein genes. The HN gene of PI2 has also been synthesized by polymerase chain reaction. In Phase II, these clones will be used to express recombinant glycoproteins in a baculovirus system; the genes will also be modified to produce secreted forms of the glycoproteins. Chimeric glycoprotein genes containing the majority of the extracellular domains of F and HN will also be constructed to produce chimeric secreted glycoproteins. The glycoproteins will be purified by immunoaffinity or lectin affinity chromatography and used for immunization of hamsters. Protection will be determined by analysis of systemic/local antibody responses and by challenge infection. The long ter objective is to produce a multivalent subunit vaccine to control human parainfluenza virus infection.

我们构建了人副流感I型(PI1)的cDNA文库 质粒载体中的2型(PI2)病毒mRNAs。两个糖蛋白基因克隆 从文库中可以看出，PI1的HN和PI2的F已经被克隆 特色化的。对其他糖蛋白基因的筛选仍在继续。这个 用聚合酶链式反应合成了PI2的HN基因。在……里面 第二阶段，这些克隆将用于表达重组糖蛋白。 杆状病毒系统；这些基因也将被修改以产生分泌物 糖蛋白的各种形式。含有嵌合糖蛋白基因的 F和HN的大部分胞外结构域也将被构建 产生嵌合分泌糖蛋白。糖蛋白将会是 通过免疫亲和层析或凝集素亲和层析纯化，用于 仓鼠的免疫接种。保护将通过分析以下内容来确定 全身/局部抗体反应和攻击感染。长发女郎 目的是生产一种多价亚单位疫苗来控制人类 副流感病毒感染。