辣椒脉斑驳病毒外壳蛋白多克隆抗体的制备和应用OA北大核心CSTPCD

[Objective]A highly specific polyclonal antibody of the capsid of chilli veinal mottle virus(ChiVMV)was prepared for field detection and quarantine inspection on infected Solanaceae plants.[Methods]The full-length(861 bp)and partial fragment(396 bp)of the gene from a ChiVMV GZ-Tabacco isolate were amplified using RT-PCR,recombined into the prokaryotic expression vector pET28a,and transformed into E.coil BL21 for induced expression.After chromatographic isolation and dialysis purification,the products were used to immunize Dahl rabbits for the antibody preparation.Potency and specificity of the candidate antibodies were evaluated by ELISA and western blot.[Results]Two polyclonal antibodies,namely antiCP1-287aa and antiCP1-132aa with the titers of 1∶6400 and 1∶12800,respectively,were obtained.Both positively detected the antigen as shown by western blot.But the indirect ELISA on the field strains revealed that antiCP1-132aa specifically detected ChiVMV not potato virus Y(PVY),whereas antiCP1-287aa failed to differentiate the two.[Conclusion]The identified polyclonal antibody,antiCP1-132aa was highly specific in detecting ChiVMV in field tests.It could become a useful tool for further studies on the infectious virus of chilli peppers.