University of Illinois Urbana-Champaign

Studies of the 50 kDa protein of barley yellow dwarf virus

Cheng, Shu-Ling

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Description

Title
Studies of the 50 kDa protein of barley yellow dwarf virus
Author(s)
Cheng, Shu-Ling
Issue Date
1995
Doctoral Committee Chair(s)
  • D'Arcy, Cleora J.
  • Domier, Leslie L.
Department of Study
  • Biology, Microbiology
  • Agriculture, Plant Pathology
Discipline
  • Biology, Microbiology
  • Agriculture, Plant Pathology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
  • Biology, Microbiology
  • Agriculture, Plant Pathology
Language
eng
Abstract
BYDVs, like all luteoviruses, are phloem-limited and are present at very low concentrations in plants. To study the readthrough protein of BYDV-PAV, the cDNA of BYDV-PAV-IL ORF 5, encoding the 50kDa protein, was cloned, expressed in E. coli by the vector pGEX-2T, and purified in a glutathione column. Polyclonal antibodies against the bacterial 50kDa fusion protein and two monoclonal antibodies, designated PAV-IL-22kDa and PAV-IL-50kDa, which are specific to CP and 50kDa protein, respectively, were produced and used in this study. The 72kDa readthrough protein of BYDV-PAV was detected in vivo (infected leaf tissue) and in vitro (purified virus) by MAb PAV-IL-50kDa. The gene expression of the 50kDa protein and its corelationship with CP expression studied by western blot, ELISA, and in situ immunogold localization assay suggest that the 50-kDa protein of BYDV is produced by readthrough of a stop codon of CP at a lower ratio, is expressed concurrently with the 22-kDa CP, accumulates mainly in the cytoplasm, but occasionally is found in the nucleus of the infected cells in the advanced stage. Studies of the competition of the purified 50kDa protein with purified virus during aphid transmission using membrane feeding techniques suggest that the 50kDa protein of BYDV-PAV may be responsible for recognition of receptors in the aphid vectors. Trypsin-digestion of BYDV-PAV for studies of aphid transmission and electron microscopy indicates that the 50kDa protein is easily degraded and the p36 domain is essential for the structural integrity of the virions. High resolution imaging of BYDV provides evidence for the presence of a structural protein protruding from the surface of the virion. This spike-like structural protein is believed to be the readthrough protein of BYDV-PAV virions.
Type of Resource
text
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http://hdl.handle.net/2142/23682
Copyright and License Information
Copyright 1995 Cheng, Shu-Ling

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