Nucleotide Sequence of the Polyprotein of the Tick—borne encephalitis virus Senzhang strain Isolated in China

Abstract：To assign the ck．borne encephalitis virus(TBEV)Senzhang strain isolated in China to one of three TBEV subtypes and understand the relation between the polyprotein structure an d biological function an d aid in the development of new TBEV vaccines．the nucleotide sequence of the polyprotein of Senzhan g strain was determined．Nine pairs of primers were designed according to the nucleotide sequence of Sofjin．HO。Oshima 5·10 strain．By RT·PCR，cDNA fragments of Senzhang strain were acquired from infected BHK．21 cells．Then the cDNA fragments was cloned into the vector pGEM ％T and then tran sforrned to competent DH5 Q cells．Positive clones were screened an d identified by EcoR I cleavage an d PCR．Th e sequences of inserted fragment were determined bv 3 1o0 Genetic Analyzer． Sequence analysis showed that Senzhang strain contains a sigle open reading frame(ORF)of 10245 nucleotides(nt)which encodes a polyprotein of 34 1 4 amino acids(aa)．Senzhang strain is more similar to Oshima5·10，So1]in·HO，So1]in，and assigned to the Far Eastern Subtype．