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

首先测定了副粘病毒、猴病毒5和41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蛋白各组分的氨基酸序列。获得了NP、P和L蛋白的全长c DNA克隆。接下来，构建了人工微基因组，即Leader Sequence-R1-Report基因-R2-Trailer Sequence。将微基因组导入人副流感病毒2型或猴病毒41型感染细胞。在该系统中，进行了复制、转录和RNA编辑。然而，效率并不是那么高。随后，将小基因组、NP、P和L蛋白的c DNA分别导入含有T7聚合酶的重组痘苗病毒感染的细胞。在这个新系统中，观察到了高效率的转录、复制和RNA编辑。在另一项实验中，几乎构建了人副流感病毒2型的全长cDNA克隆。

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.