A cloning and partial nucleotide sequencing of the amplified E gene fragment of dengue-2 virus

The E gene fragment was amplified from RNA isolated the Chinese dengue-2 virus by RT-PCR method. It was 1.29 kb in length. This unmodified E fragment was directly inserted into the EcoRV-cut T-tailed pBluescript II KS+ vector DNA. The positive recombinant colonies were identified by the PCR and the restriction endonuclease digesting method. Then nucleotide sequence analysis of the inserted E gene fragment was conducted directly by using the common M(13) primer. The result of the restriction endonuclease and sequence analysis confirmed that the 120 bp of 5'-terminal nucleotides sequence of the amplified E gene fragment was the same sequence as reported.