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UBC Theses and Dissertations
Identification and characterization of small rnas and proteins expressed by the human parvovirus B19 St. Amand, Janet Lynn
Abstract
The human pathogenic parvovirus B19 has a strict tissue tropism and will only replicate in a subset of erythroid progenitor cells. However, it is shown that COS-7 cells transfected with SV4O-B19 hybrid vectors express the major B19 RNAs and proteins. In addition, capsid proteins synthesized in these cells self-assemble into virus particles that by EM are morphologically very similar to native B19 virions. Cytoplasmic RNA from transfected COS-7 cells was used to prepare a cDNA library using a method which enriched the library for B19 cDNAs. A second cDNA library was prepared from B19 infected human bone marrow cells that had been isolated from a patient with a chronic myelogenous leukemia using PCR to amplify B19-specific cDNAs. The libraries were probed with a series of RNA probes derived from different regions of the B19 genome in pYT1O3 and selected cDNAs were sequenced and compared. Two size classes of small, abundant, polyadenylated RNAs were identified; the 700 and 800 nt size class of RNA is the product of transcriptional processing in the middle of the B19 genome downstream from an unusual polyadenylation signal, ATTAAA or AATAAC (map unit 49). These transcripts contain an ORF within their second exon which is the same reading frame used for the translation of the nonstructural proteins; however, there are no AUG translation initiation codons within this region of the ORF. The second most abundant size class of RNA in B19 infected cells are the 500 and 600 nt transcripts which are made from three exons and terminated at a normal polyadenylation signal (AATAAA) near the right hand end of the genome (map unit 97). A 94 aa ORF within the third exon is invariant in the two RNA species. [more abstract]
Item Metadata
Title |
Identification and characterization of small rnas and proteins expressed by the human parvovirus B19
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
1992
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Description |
The human pathogenic parvovirus B19 has a strict tissue tropism and
will only replicate in a subset of erythroid progenitor cells. However, it is
shown that COS-7 cells transfected with SV4O-B19 hybrid vectors express the
major B19 RNAs and proteins. In addition, capsid proteins synthesized in
these cells self-assemble into virus particles that by EM are morphologically
very similar to native B19 virions. Cytoplasmic RNA from transfected COS-7
cells was used to prepare a cDNA library using a method which enriched the
library for B19 cDNAs. A second cDNA library was prepared from B19 infected
human bone marrow cells that had been isolated from a patient with a
chronic myelogenous leukemia using PCR to amplify B19-specific cDNAs.
The libraries were probed with a series of RNA probes derived from
different regions of the B19 genome in pYT1O3 and selected cDNAs were
sequenced and compared. Two size classes of small, abundant, polyadenylated
RNAs were identified; the 700 and 800 nt size class of RNA is the product of
transcriptional processing in the middle of the B19 genome downstream from
an unusual polyadenylation signal, ATTAAA or AATAAC (map unit 49).
These transcripts contain an ORF within their second exon which is the same
reading frame used for the translation of the nonstructural proteins;
however, there are no AUG translation initiation codons within this region
of the ORF. The second most abundant size class of RNA in B19 infected cells
are the 500 and 600 nt transcripts which are made from three exons and
terminated at a normal polyadenylation signal (AATAAA) near the right
hand end of the genome (map unit 97). A 94 aa ORF within the third exon is
invariant in the two RNA species. [more abstract]
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Extent |
4499752 bytes
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Genre | |
Type | |
File Format |
application/pdf
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Language |
eng
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Date Available |
2009-01-05
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Provider |
Vancouver : University of British Columbia Library
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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.
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DOI |
10.14288/1.0086585
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
1992-05
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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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.