The whole viral genomic sequencing and phylogenetic analysis
The viral genome was amplified in fragments using specific primers
reported previously [16], and the non-coding region was amplified
according to the instructions of the 5’Full RACE and 3’Full RACE Core
Set Ver.2.0 kits (Takara Biomedical Technology Co., Ltd, Dalian). After
each fragment was ligated into the pMD18-T vector, Escherichia
coli DH5α cells were transformed with recombinant plasmids. All the
fragments were sequenced by Sangon Biotech Co., Ltd (Shanghai).
The sequenced fragments were spliced using DNAMAN software. Sequence
similarity and phylogenetic analyses were conducted using DNAStar and
MEGA4.0. Furthermore, the amino acid sequences of GETV E2 genes were
compared and homology was analyzed using MegAlign. Phylogenetic analysis
of amino acid sequences derived from the E2 gene were conducted using
neighbor-joining methods (1000 bootstrap replicates) in MEGA4.0.