The Plant Pathology Journal 2009;25(3):225-230.
Published online September 30, 2009.
Production and Evaluation of Monoclonal Antibodies Against Recombinant Coat Protein of Lily mottle virus for Western Blotting and Immono-blot Analysis
Bong Nam Chung, Ju Yeon Yoon, Gug Sun Choi
Abstract
Lily mottle virus (LMoV) causes flower quality reduction in Lilium spp. The coat protein gene was RT-PCRamplified from total RNA extracted from infected lily leaves and the amplified fragment was cloned into the pRSET expression vector tagged with a His-MBP. The plasmid of recombinant coat protein was used to transform an Escherichia coli strain pLysS and was expressed. The coat protein was purified by affinity chromatography using a Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE. The in vitroexpressed protein was used for immunization of mice. The polyclonal and monoclonal antibodies reacted specifically for the detection of LMoV in lily extracts in Western blot. Moreover the monoclonal antibodies reacted with lily extracts in DAS-ELISA with no unspecific or heterologous reactions against other nonserologically related viruses, but the polyclonal antibodies revealed a weak reaction against both infected lily and healthy control.
Key Words: Lily mottle virus, monoclonal antibody coat protein, lily
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