[Cloning and primary structure of DNA copies of genome fragments of the tick-borne encephalitis virus].

RNA of a flavivirus-tick-borne encephalitis virus (Far-East, type 1, strain Sofin) was subjected to reverse transcription and the DNA copy was transformed into double-stranded DNA by action of E. coli DNA-polymerase (Klenow's fragment) without primer. The hairpin structures were removed by S1 nuclease. Oligo-dC ends were attached to ds-cDNA thus obtained, and this DNA was annealed with pBR322 plasmid cut by PstI and equipped with oligo-dG termini. The recombinant plasmids were cloned in E. coli HB101. Of the 360 TcrAps clones obtained, 187 clones efficiently hybridized with partially degraded 32P-RNA of TBE virus. The sequence of the insert of one of the clones was determined by the Maxam-Gilbert method. The 720 b.p. sequence is translatable into an amino acid sequence without interruption. Nearby the 3'-terminus of the insert, the sequence ACACAGG is present which is homologous with that found in RNA of the flavivirus West Nile.