Abstract: Sugarcane mosaic disease caused by Sorghum mosaic virus(SrMV)is now recognized as one of the most widely distributedviral diseases of Saccharum. It is an important limiting factor for sugarcane production. According to the sequence of coat protein(CP)gene ofSrMV, two primers were designed, and a DNA fragment about 987 bp was amplified by RT-PCR. The PCR product, after digested with restrictionenzymes, was inserted into prokaryotic expression vector pET32a and the recombinant plasmid pET32a-SrMVCP was transferred into E.coliRosetta(DE3). Induction with IPTG, recombinant protein was got and SDS-PAGE analysis and western blot showed that CP was expressedsteadily in soluble proteins. With addition of IPTG up to 0.1 mmol/L and culturing for 4 h more in 37℃ is thought to be the best inducingconditions to express CP. The fusion protein was then purified with Ni2+-NTA agarose affinity chromatography, and used to immune rabbits forantiserum preparation. The optimal titer of the antiserum was determined to be 1∶1 000 tested by indirect enzyme-linked immunosorbent assay(ID-ELISA). Western blotting confirmed that the antiserum reacted strongly and specifically to the CP of SrMV-HN field sugarcane sampleswere detected using the antiserum by a methed of ID-ELISA.

Key words: Sorghum mosaic virus , Coat protein , Prokaryotic expression , Antiserum