PARAINFLUENZA VIRUS TYPE 3 (PIV3) GENE EXPRESSION

副流感病毒 3 型 (PIV3) 基因表达

• 批准号: 3746506
• 负责人: P L COLLINS
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3746506
• 关键词: Paramyxovirus; chloramphenicol acetyltransferase; complementary DNA; gene expression; helper virus; microorganism culture; recombinant DNA; recombinant proteins; transfection; viral rescue; virus genetics

Previously, we developed an experimental "rescue" system for human parainfluenza virus type 3 (PIV3) based on a cDNA-encoded minigenome, PIV3-CAT. This is a truncated version of PIV3 genome RNA in which all of the viral genes have been removed and replaced by the bacterial chloramphenicol acetyl transferase (CAT) gene. When synthesized in vitro and transfected into cells which have been infected with standard PIV3 helper virus, PIV3-CAT was rendered biologically active ("rescued") as evidenced by the intracellular expression of CAT and by the production of a component (presumably infectious particles) that was released into the medium and could be passed productively onto fresh cells in the presence of helper virus. Together with a similar system developed by us for human respiratory syncytial virus (RSV) (accompanying report) and by others for Sendai virus, this represents the first experimental method by which one can synthesize, and thereby manipulate, biologically-active analogs of the genome of a nonsegmented negative strand virus. It will now be possible to directly identify and characterize cis-acting signals in genome and, as described here, considerable progress has been made in this area. The success in rescuing PIV3-CAT supports the idea that it will be possible to adapt the system such that a complete synthetic genome can be used to produce nondefective virus. This would make possible the creation of viruses that contain defined mutations for vaccine and experimental purposes.

以前，我们开发了人类的实验“救援”系统 基于cDNA编码的微型素组的parainfluenza病毒3型（PIV3）， piv3-cat。 这是PIV3基因组RNA的截断版本，其中所有版本 病毒基因已被清除并被细菌取代 氯霉素乙酰转移酶（CAT）基因。 当体外合成时 并转染已感染标准PIV3的细胞 辅助病毒，PIV3-CAT被赋予生物活性（“救出”）为 由CAT的细胞内表达和生产证明 被释放到的成分（可能是传染性颗粒） 培养基可以有效地传递到新鲜细胞中 辅助病毒的存在。 与类似的系统一起开发 美国呼吸道合胞病毒（RSV）（随附的报告）和 其他人的仙台病毒，这代表了第一种实验方法 通过它可以合成，从而操纵生物学活性 非段负链病毒基因组的类似物。 会 现在可以直接识别和表征顺式作用信号 在基因组中，如下所述，在 这个区域。 营救PIV3-CAT的成功支持了这样的想法 可以调整系统以使完整的合成 基因组可用于产生非缺陷病毒。 这会做到 可能创建包含定义突变的病毒 疫苗和实验目的。