2008•Huazhong Nongye Daxue xuebaoRequires access
Construction of Prokaryotic Vector and Expression of the CP Fragment of Tobacco ringspot virus
Yanyan Qiao
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Abstract
The CP fragment of the Tobacco ringspot virus(TRSV) was amplified by RT-PCR,and inserted into the expression vector pET28a.The recombinant plasmid pET-CP was transformed into Escherichia coli BL21(DE3),the target protein was expressed with induction of IPTG and purified by Ni+-NTA metal chelation chromatography.The purified recombinant protein was analyzed by SDS-PAGE and Western blotting,the results showed that the CP fragment was highly expressed.The molecular weight of the recombinant protein was about 34 ku,which was immunological reactive.The product was used as antigen to immunize the rabbit,and the antiserum had a titer of 1∶409 600,it was reaction between antiserum and recombinant protein.