摘要
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.
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 Sal I and BamH I, 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.
出处
《中国病毒学》
CSCD
2006年第1期78-80,共3页
Virologica Sinica
基金
黑龙江省政府博士后科研启动金资助项目(LRB-KY01045)
