E.coli Expression of Equine West Nile Virus Partly E Protein

Abstract:

According to the GenBank published sequence of equine west nile virus(WNV)E protein gene,a pair of primer was designed in order to amplify equine WNV partly E gene by RT-PCR.The fragment was 318bp in length and was cloned into pMD18-T-Vector.The positive clone was named pMD-E and was sequenced.Then it was sub-cloned into pET-32a(+).The recombinant pET32a-E plasmid, which includes the gene fragment of the equine WNV E protein,was transformed into E.coli BI21,and expressed about 33.1%.The ex- pressed product was about 32kD molecular weight by SDS-PAGE.The purified product could be recognized by positive serum of WNV by Wester-blotting. It was laid a foundation for developing an ELISA to detect equine WNV used the purified production as coated antigen.

Key words:Equine west nile virus; E protein gene; Expression