Zhang H, Air G M
Department of Microbiology, University of Alabama at Birmingham 35294.
Biochem Biophys Res Commun. 1994 Apr 15;200(1):95-101. doi: 10.1006/bbrc.1994.1419.
cDNAs containing the coding sequences of influenza type A virus polymerase proteins (PB1, PB2 and PA) and nucleoprotein (NP) have been expressed in mammalian cells by T7 polymerase provided by a recombinant vaccinia virus. The resulting proteins are able to form a complex that can copy a negative sense influenza-like RNA, transcribed from input DNA by the T7 polymerase, into a positive sense RNA that is translated into active chloramphenicol acetyltransferase (CAT). In this system there is no requirement for helper virus or purified viral core proteins, thus it will allow manipulation of all proteins as well as template for studies of replication in influenza virus.
含有甲型流感病毒聚合酶蛋白(PB1、PB2和PA)及核蛋白(NP)编码序列的cDNA,已通过重组痘苗病毒提供的T7聚合酶在哺乳动物细胞中表达。所产生的蛋白质能够形成一种复合物,该复合物可将由T7聚合酶从输入DNA转录而来的负链流感样RNA复制成正链RNA,进而翻译为活性氯霉素乙酰转移酶(CAT)。在该系统中,不需要辅助病毒或纯化的病毒核心蛋白,因此它将允许对所有蛋白质以及用于流感病毒复制研究的模板进行操作。