- Library Home /
- Search Collections /
- Open Collections /
- Browse Collections /
- UBC Theses and Dissertations /
- The complete nucleotide sequence of prune dwarf ilarvirus...
Open Collections
UBC Theses and Dissertations
UBC Theses and Dissertations
The complete nucleotide sequence of prune dwarf ilarvirus RNA1 and virus detection by reverse transcription PCR and triple-antibody sandwich ELISA Rampitsch, Christof
Abstract
A triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) with a monoclonal antibody was developed and evaluated for the detection of prune dwarf ilarvirus (PDV) in sweet cherry trees {Prunus avium). A reverse transcribed polymerase chain reaction test was also developed to establish the incidence of PDV in 40 sweet cherry trees and to confirm the absence of virus in 15 control trees. Trees with two-thirds of their leaves positive for PDV by TAS-ELISA would be identified with 99% probability by testing four leaves per tree. The monoclonal antibody did not cross- react with Prunus necrotic ringspot ilarvirus in the TASELISA. The nucleotide sequence of PDV RNA1 was determined. The RNA consists of 3374 nucleotides and encodes a single open reading frame of 3168 nucleotides. The putative translation product is 1055 amino acids in length with a calculated molecular mass of 118.9 kDa. Both the nucleic acid and the translated amino acid sequences show stronger homology to RNA1 and the corresponding translation product (ORF1) of alfalfa mosaic alfamovirus (AMV) than' to citrus leaf rugose ilarvirus, the only other ilarvirus for which RNA1 sequence data is available. There is extensive sequence homology in the 3'-untranslated regions of PDV RNA1 and the 3'-regions of other ilarvirus and AMV RNAs. The reported sequence and its single open reading frame conform to the genomic organization typical of the Bromoviridae genus. Clones representing sequence from the 5'and 3'-end of RNA1 were used to construct a deletion-type defective interfering particle and its ability to replicate in vivo was assessed.
Item Metadata
| Title |
The complete nucleotide sequence of prune dwarf ilarvirus RNA1 and virus detection by reverse transcription PCR and triple-antibody sandwich ELISA
|
| Creator | |
| Publisher |
University of British Columbia
|
| Date Issued |
1996
|
| Description |
A triple antibody sandwich enzyme-linked immunosorbent
assay (TAS-ELISA) with a monoclonal antibody was developed
and evaluated for the detection of prune dwarf ilarvirus
(PDV) in sweet cherry trees {Prunus avium). A reverse
transcribed polymerase chain reaction test was also developed
to establish the incidence of PDV in 40 sweet cherry trees
and to confirm the absence of virus in 15 control trees.
Trees with two-thirds of their leaves positive for PDV by
TAS-ELISA would be identified with 99% probability by testing
four leaves per tree. The monoclonal antibody did not cross-
react with Prunus necrotic ringspot ilarvirus in the TASELISA.
The nucleotide sequence of PDV RNA1 was determined. The
RNA consists of 3374 nucleotides and encodes a single open
reading frame of 3168 nucleotides. The putative translation
product is 1055 amino acids in length with a calculated
molecular mass of 118.9 kDa. Both the nucleic acid and the
translated amino acid sequences show stronger homology to
RNA1 and the corresponding translation product (ORF1) of
alfalfa mosaic alfamovirus (AMV) than' to citrus leaf rugose
ilarvirus, the only other ilarvirus for which RNA1 sequence
data is available. There is extensive sequence homology in
the 3'-untranslated regions of PDV RNA1 and the 3'-regions of
other ilarvirus and AMV RNAs. The reported sequence and its
single open reading frame conform to the genomic organization typical of the Bromoviridae genus.
Clones representing sequence from the 5'and
3'-end of
RNA1 were used to construct a deletion-type defective
interfering particle and its ability to replicate in vivo was
assessed.
|
| Extent |
6661533 bytes
|
| Genre | |
| Type | |
| File Format |
application/pdf
|
| Language |
eng
|
| Date Available |
2009-03-17
|
| Provider |
Vancouver : University of British Columbia Library
|
| Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
|
| DOI |
10.14288/1.0087819
|
| URI | |
| Degree | |
| Program | |
| Affiliation | |
| Degree Grantor |
University of British Columbia
|
| Graduation Date |
1996-11
|
| Campus | |
| Scholarly Level |
Graduate
|
| Aggregated Source Repository |
DSpace
|
Item Media
Item Citations and Data
Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.