Completion of Kunjin virus RNA sequence and recovery of an infectious RNA transcribed from stably cloned full-length cDNA

摘要: Completion of the Kunjin virus (KUN) RNA sequence showed that it is longest flavivirus reported (11,022 bases), commencing with a 5' noncoding region 96 bases. The 3' 624 nucleotides included unique insertion 46 bases adjacent to stop codon, but otherwise had properties similar those RNAs closely related flaviviruses. A full-length KUN cDNA clone which could be stably propagated in Escherichia coli DH5 alpha was constructed; SP6 polymerase transcripts from amplified were infectious when transfected into BHK-21 cells. mutational change abolishing BamHI restriction site at position 4049, leading conservative amino acid Arg-175 Lys NS2A protein, introduced during construction and retained recovered virus. Extra terminal cloning shown present vitro absent Although differed parental by smaller plaque phenotype delayed growth rate cells mice, very as assessed several other criteria, such peak titer cells, infectivity adsorption penetration replication 39 degrees C, neurovirulence after intraperitoneal injection mice. cloned will provide useful basis for future studies defining characterizing functional roles all gene products.