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Native Electrophoresis and Western Blot Analysis: Method and Applications

  • Ioannis N. Manoussopoulos
  • Mina Tsagris
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 536)

Summary

Native electrophoresis and western blot analysis (NEWeB) has been developed for the study of plant virus characteristics, among others, virus particle–protein interactions, electrophorotype formation, and strain separation. The method is based on the property of electrophoretic mobility of virus particles (VP) and proteins and combines the analytical capacity of electrophoresis with the specificity of western blot. One of its advantages is that it deals with entire VP that can be studied in cause and effect or in time-interval experiments. Some of the most interesting approaches include VP structural studies, VP interaction with host or viral proteins, and also the characterization of VP–protein complexes. In this protocol, NEWeB is used to demonstrate the interaction of Plum pox virus particles with the helper component, a virus encoded protein. It is expected that the method could be used in analogous studies of other viruses or large protein complexes, where similar principles apply.

Key words

Native electrophoresis NEWeB Virus particle–protein interaction Helper component HCPro Coat protein Plum pox virus Potyvirus 

Notes

Acknowledgements

We thank Prof. E. Maiss for providing PPV CP and HC specific antibodies and M. Bastaki for help during her rotation. This work was partly supported by the Greek Ministry of Development (Project EPET II, 453) and by the INTAS 989 program of the EC.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.National Agricultural Research Foundation, Plant Protection Institute of Patras, Plant Pathology DivisionLaboratory of VirologyNEO & AmerikisGreece

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