Submitted to: Workshop on Molecular Pathogenesis of Marek's Disease and Avian Immunology
Publication Type: Abstract Only
Publication Acceptance Date: August 5, 2006
Publication Date: August 5, 2006
Citation: MacLea, K.S., Cheng, H.H. 2006. Purification of MDV-chicken protein complexes using molecularly cloned and tagged MDV-BACS [abstract]. 4th International Workshop on Molecular Pathogenesis of Marek's Disease Virus. p. 38. Technical Abstract: With an eye to developing chicken lines resistant to MDV, work in our laboratory has integrated several genomic approaches, including virus-host protein-protein interaction screens, into a comprehensive program for identifying putative host resistance genes. Previous efforts made use of a two-hybrid screen to identify viral proteins interacting with a library of chicken gene products in bacteria. In the current set of experiments, we have chosen to examine MDV-chicken protein-protein interactions in a more natural context. Specifically, the recent generation of several infectious MDV-BAC clones has made possible the easy manipulation of the virus to generate defined recombinant MDVs. Furthermore, making use of developments in gentle protein complex extraction and identification, we have developed recombinant MDV-BAC clones in which each of several open reading frames (ORFs) are expressed as fusion products with tandem affinity purification (TAP) tags. These TAP tags allow virus-host protein complexes from cultured chicken cells to be recovered from whole cell lysates under near-physiological conditions. Through the use of liquid chromatography-tandem mass spectrometry technology, these studies, when completed, will allow identification of viral and host proteins interacting with each MDV ORF and provide targets for further examination of the role of these proteins in resistance to viral infection and/or pathogenesis.