INVESTIGATION OF REPLICATION MECHANISM AND PATHOGENESIS OF PARAMYXOVIRUS BY USING INFECTIOUS cDNA CLONE

首先测定了副粘病毒、类人猿病毒5号和41号的L基因的核苷酸序列。因此，还测定了复制复合体、NP、P和L蛋白组分的氨基酸序列。获得了NP、P和L蛋白的全长cDNA克隆。然后构建Leader序列- r1报告基因- r2 - trailer序列的人工微基因组。将这些微型基因组转染到人副流感2型病毒或猿猴41型病毒感染的细胞中。在该系统中进行复制、转录和RNA编辑。然而，效率并没有那么高。随后，将微基因组、NP、P和L蛋白cDNA转染到含有T7聚合酶的重组痘苗病毒感染的细胞中。该系统具有较高的转录、复制和RNA编辑效率。在另一个实验中，人类副流感病毒2型全长cDNA克隆已基本构建。

At first, the nucleotide sequences of the L gene of Paramyxovirus, simian viruses 5 and 41 were determined. Therefore, amino acid sequences of components of replicative complex, NP, P, and L proteins were also determined. Full-length cDNA clones for NP,P, and L protein could be obtained. Next, artificial mini-genomes, Leader sequence-R1-reporter gene-R2-Trailer sequence, were constructed. The mini-genomes were transfected into human parainfluenza type 2 virus or simian virus 41-infected cells. In this system, replication, transcription and RNA editing were occurred. However, the efficiency was not so high. Subsequently, the mini-genomes, cDNA for NP, P, and L proteins were transfected into the cells infected recombinant vaccinia virus containing T7 polymerase. In this new system, high efficiency of transcription, replication and RNA editing were observed. In another experiment, full-length cDNA clone for human parainfluenza virus type 2 was nearly constructed.