Preparation of Adenovirus 5 E1A Polyclonal Antibody

来源 :Chemical Research in Chinese Universities | 被引量 : 0次 | 上传用户:zxj3510
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The adenovirus E1A proteins are primarily concerned with modulating cellular metabolism to make the cell more susceptible to virus replication. In this study, adenovirus E1A gene was amplified via PCR on the HEK-293 cells genome and cloned into prokaryotic expression vector pRsetB. The recombinant plasmid pRsetB-E1A was expressed in E.coli BL21 and the relative molecular mass(Mr) of expressed fusion protein was approximately 36000. The recombinant protein was purified on a Ni-NIA agarose column and detected by SDS-PAGE and Western blot. The purified recombinant protein was then injected into rabbits and anti-E1A polyclonal antibody with high titer was obtained. The adenovirus E1A proteins are primarily concerned with modulating cellular metabolism to make the cell more susceptible to virus replication. In this study, adenovirus E1A gene was amplified via PCR on the HEK-293 cells genome and cloned into prokaryotic expression vector pRsetB. The recombinant plasmid pRsetB-E1A was expressed in E. coli BL21 and the relative molecular mass (Mr) of expressed fusion protein was approximately 36000. The recombinant protein was purified on a Ni-NIA agarose column and detected by SDS-PAGE and Western blot. The purified protein was purified recombinant protein was then injected into rabbits and anti-E1A polyclonal antibody with high titer was.
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