Luytjes W, Krystal M, Enami M, Parvin J D, Palese P
Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029.
Cell. 1989 Dec 22;59(6):1107-13. doi: 10.1016/0092-8674(89)90766-6.
A system is described that allows use of recombinant DNA technology to modify the genome of influenza virus, a negative-strand RNA virus, and to engineer vectors for the expression of foreign genes. Recombinant RNA is expressed from plasmid DNA in which the coding sequence of the influenza A virus NS gene is replaced with that of the chloramphenicol acetyltransferase gene. When transfected with purified influenza A virus polymerase proteins--in the presence of helper virus--the recombinant RNA is amplified, expressed, and packaged into virus particles, which can be passaged several times. The data indicate that the 22 5' terminal and the 26 3' terminal bases of the influenza A virus RNA are sufficient to provide the signals for RNA transcription, RNA replication, packaging of RNA into influenza virus particles.
本文描述了一种系统,该系统允许使用重组DNA技术来修饰负链RNA病毒——流感病毒的基因组,并构建用于表达外源基因的载体。重组RNA由质粒DNA表达,其中甲型流感病毒NS基因的编码序列被氯霉素乙酰转移酶基因的编码序列所取代。当在辅助病毒存在的情况下用纯化的甲型流感病毒聚合酶蛋白转染时,重组RNA被扩增、表达并包装成病毒颗粒,这些病毒颗粒可以传代多次。数据表明,甲型流感病毒RNA的22个5'末端碱基和26个3'末端碱基足以提供RNA转录、RNA复制以及将RNA包装到流感病毒颗粒中的信号。