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 wit...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.展开更多
文摘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.
