Amplification And Cloning Of Full-length JEV CDNA And Preparation Of Infectious Transcripts By In Vitro Transcription

Japanese encephalitis virus belongs to the family Flaviviridae with a genome of capped single-strand positive RNA approximately 11kb in length. The virus causes severe encephalitic and neurologic disease manifestations. Among flaviviruses, JEV infection has the highest mortality with more than 10,000 deaths annually. Studies on its replication and virulence at gene level will facilitate the development of vaccine and the understanding of pathogenic mechanism. However the study of molecular biology of nonretroviral RNA viruses has long been hampered by the fact that these viruses do not encompass a DNA intermediate step in their replication cycle, since to date extremely varied and powerful molecular biology techniques aimed at modifying nucleic acids have been directly at DNA substrates. The technique of preparation of transcripts by in vitro transcription from full-length cDNA corresponding to virus RNA greatly enhanced the molecular studies of RNA viruses. The procedure includes:(1) placing full-length cDNA of virus directly...