Kimura N, Nishida M, Nagata K, Ishihama A, Oda K, Nakada S
Department of Biological Science and Technology, Science University of Tokyo, Chiba, Japan.
J Gen Virol. 1992 Jun;73 ( Pt 6):1321-8. doi: 10.1099/0022-1317-73-6-1321.
A new transfection system for influenza virus was developed using the clone 76 cell line, in which the viral RNA polymerase and nucleoprotein (NP) genes can be expressed in response to dexamethasone. Ribonucleoprotein (RNP) complexes were reconstituted by expressing proteins from a chimeric NS-chloramphenicol acetyltransferase (CAT) RNA consisting of the full-length negative-strand RNA of the CAT gene positioned between the 5'- and 3'-terminal sequences of influenza virus RNA segment 8, and purifying NP from an NP gene-expressing Escherichia coli strain. When the reconstituted RNP was transfected into clone 76 cells, CAT was produced only when the synthesis of the three RNA polymerase subunits and NP was induced by treatment with dexamethasone.
利用克隆76细胞系开发了一种新型流感病毒转染系统,在该系统中,病毒RNA聚合酶和核蛋白(NP)基因可响应地塞米松表达。核糖核蛋白(RNP)复合物通过表达来自嵌合NS-氯霉素乙酰转移酶(CAT)RNA的蛋白质来重建,该RNA由位于流感病毒RNA片段8的5'-和3'-末端序列之间的CAT基因全长负链RNA组成,并从表达NP基因的大肠杆菌菌株中纯化NP。当将重建的RNP转染到克隆76细胞中时,只有在用DEX处理诱导三种RNA聚合酶亚基和NP的合成时才产生CAT。