University of Illinois Urbana-Champaign

Characterization and nucleotide sequence of apple mosaic virus coat protein gene

Alrefai, Rudaina Hussein

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https://hdl.handle.net/2142/21141

Description

Title
Characterization and nucleotide sequence of apple mosaic virus coat protein gene
Author(s)
Alrefai, Rudaina Hussein
Issue Date
1993
Doctoral Committee Chair(s)
Korban, Schuyler S.
Department of Study
Biology
Discipline
Biology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
  • Agriculture, Agronomy
  • Biology, Molecular
  • Agriculture, Plant Pathology
Language
eng
Abstract
  • Apple mosaic virus (ApMV) (Idaho strain) was purified from infected cucumber cotyledons. Virions 3 and 4 (top components in sucrose gradient) were isolated and RNA was extracted. A double stranded copy DNA (cDNA) was synthesized from in vitro polyadenylated RNA3 and cloned into a pBluescript T-vector. The inserts of four clones from the 3$\sp\prime$-end of RNA3 were sequenced and the 3$\sp\prime$-primer was designed to synthesize cDNA of RNA3. The DNA was cloned into the plasmid vector pUC118. A set of overlapping cDNA clones was used to determine the nucleotide sequence of RNA4. The 5$\sp\prime$-end of RNA4 was mapped using the primer extension technique. ApMV RNA4 was found to be 983 nucleotides in length containing open reading frame (ORF) of a 662 nucleotides, which was flanked by 5$\sp\prime$ and 3$\sp\prime$-nontranslated sequence of 55 and 265 nucleotides, respectively. The ORF encodes for a coat protein of a 25,085 Mr. The sequence contains several inverted repeats that could form hair-pin structures. In addition, two palindromic sequence were found in the ORF with a third palindrome in the 3$\sp\prime$-nontranslated region. These structures may be involved in the recognition of RNA by viral replicase or in genome activation by the coat protein. The amino-terminus of ApMV coat protein is basic and rich in Cys, whereas the carboxy-terminus has an even distribution of basic and acidic amino acids, similar to other ilarviruses.
  • "To transform plants with ApMV coat protein gene an Agrobacterium-mediated transformation system was developed for apple ""Royal Gala"" plants. Kanamycin completely inhibited regeneration of untransformed leaf sections on a 1/2 Murashige and Skoog (MS) regeneration medium. Apple leaf sections were transformed by Agrobacterium strains C58Z707:pZA-7 and EHA101:pZA-7 and calli were produced on a 1/2MS selection medium."
Type of Resource
text
Permalink
http://hdl.handle.net/2142/21141
Copyright and License Information
Copyright 1993 Alrefai, Rudaina Hussein

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