The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Production and Evaluation of Monoclonal Antibodies Against Recombinant Coat Protein of Lily mottle virus for Western Blotting and Immono-blot Analysis

  • Chung, Bong-Nam (National Institute of Horticultural and Herbal Science, Rural Development Administration) ;
  • Yoon, Ju-Yeon (Plant Virus GenBank, Division of Environmental Life Science, Seoul Women's University) ;
  • Choi, Gug-Sun (National Institute of Horticultural and Herbal Science, Rural Development Administration)
  • Published : 2009.09.30
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Abstract

Lily mottle virus (LMoV) causes flower quality reduction in Lilium spp. The coat protein gene was RT-PCR-amplified 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 vitro-expressed 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 non-serologically related viruses, but the polyclonal antibodies revealed a weak reaction against both infected lily and healthy control.

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References

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