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Title:Studies of the 50 kDa protein of barley yellow dwarf virus
Author(s):Cheng, Shu-Ling
Doctoral Committee Chair(s):D'Arcy, Cleora J.; Domier, Leslie L.
Department / Program:Biology, Microbiology
Agriculture, Plant Pathology
Discipline:Biology, Microbiology
Agriculture, Plant Pathology
Degree Granting Institution:University of Illinois at Urbana-Champaign
Subject(s):Biology, Microbiology
Agriculture, Plant Pathology
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.
Issue Date:1995
Rights Information:Copyright 1995 Cheng, Shu-Ling
Date Available in IDEALS:2011-05-07
Identifier in Online Catalog:AAI9543554
OCLC Identifier:(UMI)AAI9543554

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