Expression of Avian Infectious Bronchitis Virus E Protein in E.coli
Abstract: Reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the Small Envelope (E) gene of avian Infectious bronchitis virus (IBV) LX4 strain and the gene was cloned into the pMD18-T vector. By digestion with restriction enzymes SalⅠand BamHⅠ, the E gene was subcloned into pET-30a vector to construct recombinant plasmid pET-30a-E. The recombinant plasmid was transformed into E. coli BL21(DE3) and induced with IPTG. It was demonstrated by SDS-PAGE that a protein of 14kDa, which was comparable in size to the native E protein, was expressed in E. coli. The 14KDa protein was purified and used to prepare the rabbit antiserum against IBV E protein. The result showed that the antibody could react with purified E protein in Western blotting.