Cloning and characterization of the 3' terminal regions of RNA from select strains of maize dwarf mosaic virus and sugarcane mosaic virus

Double-stranded cDNAs derived from the 3$\sp\prime$ terminus of maize dwarf mosaic virus, strain A (MDMV-A), maize dwarf mosaic virus, strain B (MDMV-B), maize dwarf mosaic virus, strain KS1 (MDMV-KS1), maize dwarf mosaic virus, strain O (MDMV-O), and sugarcane mosaic virus strain H have been cloned as a first step in the creation of transgenic MDMV and SCMV resistant maize plants. These cDNA clones contain cDNA inserts derived from varying lengths of the coding region of the nuclear inclusion II protein, the complete coat protein cistron, and the complete untranslated region of these viruses plus a poly-adenylated 3$\sp\prime$-terminal tail. Nucleotide sequence analyses of each of these cDNA inserts have been determined. Homology studies carried out on the resultant ds DNA sequences and derived amino acid sequences show regions of high homology in comparison to one another and to previously published potyvirus nuclear inclusion II and coat protein amino acid sequences. Amino acid homologies of the derived amino acid sequences show the N-termini of the coat proteins of these viruses exhibit considerable variability and support immunological data establishing four distinct groups within the MDMV and SCMV viruses. Homology studies identified the site of proteolytic cleavage between the nuclear inclusion II proteins and the coat proteins of these viruses. Additionally, oligodeoxynucleotide directed site specific mutagenesis alteration of the MDMV-B cDNA clone allows excision of the NI$\sb{\rm II}$ coding region and introduction of a translation start site for the (MDMV-B) coat protein. Subsequent in vitro transcription and translation yields immunologically distinct MDMV-B coat protein. This thesis presents the first step in the potential generation of transgenic maize plants that express MDMV and SCMV coat proteins and exhibit resistance to infection by various strains of MDMV and SCMV.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2011

Author dc:creator

• Jilka, Joseph M.

Contributors dc:contributor

• Clark, John M., Jr.

• Biology, Molecular
• Agriculture, Plant Pathology
• Chemistry, Biochemistry

Statement dc:rights

• Copyright 1990 Jilka, Joseph M.

Language dc:language

eng

Identifier

AAI9114282
(UMI)AAI9114282

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/21792