|Chaouch, R - OHIO STATE UNIVERSITY|
|Willie, K - OHIO STATE UNIVERSITY|
|Hogenhout, S - OHIO STATE UNIVERSITY|
Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: March 13, 2002
Publication Date: August 12, 2002
Citation: Chaouch, R., Willie, K., Redinbaugh, M.G., Hogenhout, S.A. 2002. Maize chlorotic dwarf virus genome sequence and polyprotein cleavage. Phytopathology. 92:514 Technical Abstract: The genomic sequence (11.8 kb) of the severe Ohio Maize chlorotic dwarf virus isolate (MCDV-S, genus Waikavirus) was determined from overlapping cDNA clones. Approximately 400 kDa polyprotein encoded by the viral genome is post-translationally cleaved into several smaller functional proteins. Whereas the three MCDV capsid proteins, a cysteine protease and RNA-dependent RNA polymerase were identified on the polyprotein sequence, functional proteins from three separate regions of the virus polyprotein have not been identified and characterized. cDNAs for these polyprotein regions were cloned into the pTrcHis expression vector. His-tagged fusion proteins of 75.5 kDa (MCDV1), 34.6 kDa (MCDV2) and 66.4 kDA (MCDV3) proteins were produced in Escherichia coli. Antisera raised to MCDV1, 2 and 3 were used to determine the sizes of functional proteins by Western blot analysis, and to immunoprecipitate the functional proteins for subsequent N amino-terminal sequencing. Preliminary data show that MCDV1 antisera detect 50 and 35 kDa proteins in both infected plants and virions, but not in the healthy plants.