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Characterisation of new full-length HIV-1 subtype D viruses from South Africa

Thesis (MSc (Medical Virology )--University of Stellenbosch, 2004.

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Main Authors: Loxton, Andre Gareth, Janse van Rensburg, E., Engelbrecht, S.
Format: Thesis
Language:English
Published: 2010
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access_status_str Open Access
author Loxton, Andre Gareth
Janse van Rensburg, E.
Engelbrecht, S.
author_browse Engelbrecht, S.
Janse van Rensburg, E.
Loxton, Andre Gareth
author_facet Loxton, Andre Gareth
Janse van Rensburg, E.
Engelbrecht, S.
author_sort Loxton, Andre Gareth
collection Thesis
description Thesis (MSc (Medical Virology )--University of Stellenbosch, 2004.
format Thesis
id oai:scholar.sun.ac.za:10019.1/3266
institution Stellenbosch University (South Africa)
language English
last_indexed 2026-06-10T12:45:24.995Z
license_str Not specified — see source repository
provenance_str_mv Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository
publishDate 2010
publishDateRange 2010
publishDateSort 2010
record_format dspace
source_str SUNScholar — Stellenbosch University Repository
spelling oai:scholar.sun.ac.za:10019.1/3266 Characterisation of new full-length HIV-1 subtype D viruses from South Africa Loxton, Andre Gareth Janse van Rensburg, E. Engelbrecht, S. HIV-1 Homosexual transmission Virusses Epidemics DNA Cloning Glycosilation Evolutionary phylogenetic trees Tygerberg HIV-1 subtype D strains HIV infections -- Transmission -- Prevention -- South Africa DNA viruses -- South Africa Thesis (MSc (Medical Virology )--University of Stellenbosch, 2004. 150 leaves printed on single pages, preliminary pages i-vii and numberd pages 1-143. Includes bibliography and figures digitized at 300 dpi grayscale and 300 dpi 24-bit Color to pdf format (OCR), using a Hp Scanjet 8250 Scanner and digitized at 600 dpi grayscale to pdf format (OCR), using a Bizhub 250 Konica Minolta Scanner. ENGLISH ABSTRACT: The first episode of HIV-1 in South Africa was documented in 1982. Homosexual transmission of the virus was the predominate mode of transmission in an epidemic of mainly HIV-1 subtype Band D infections. To date, no full-length sequences of Subtype D strains from South Africa has been reported. Here we describe the characterization and some of the unique features of the Tygerberg HIV-1 subtype D strains. A near full-length 9 kb fragment was obtained through a one step PCR using high molecular weight DNA. Cloning was done successfully with the pCR-XLTapa cloning kit. Large quantities of plasmid DNA was grown and sequenced on both strands of the DNA. ORF determination and subtyping was followed by standard phylogenetic methods to construct evolutionary phylogenetic trees. Subtyping and similarity plots revealed that the sequences from Tygerberg are pure subtype D. All the Tygerberg strains had intact genes with no premature stop codons. At the tip of the V3 loop, the Tygerberg strains have the GOGO motif. R214 has a more variable vpu gene than the rest of the Tygerberg strains, but is still subtype D in this region. No premature stop codons have been observed in the tat gene and the glycosilation of the strains are less than the subtype D consensus. We are the first to report full-length sequences of HIV-1 subtype D strains from South Africa. The sequences represent non-mosaic genomes of subtype D. Our results confirm that the subtype D sequences from the beginning of the HIV-1 epidemic differ from the Subtype D sequences from recent isolates. AFRIKAANSE OPSOMMING: Die eerste episode van HIV-1 infeksie in Suid Afrika is in 1982 gedokumenteer. Die epidemie het hoofsaaklik uit subtipe B en D bestaan en was deur homoseksuele kontak oorgedra. Geen vollengte subtipe D DNS volgordes van Suid-Afrika is tans beskryf nie. Hier beskryf ons die karakterisering van vollengte subtipe D stamme asook sommige van die unieke eienskappe van die virusse. Die vollengte 9 kb genoom volgorde was verkry deur 'n eenstap PKR reaksie met hoë molekulêre gewig DNS uit te voer. Die 9 kb fragment was suksesvol gekloneer met behulp van die peR-Xl-TOPO klonerings toetsstel. Groot hoeveelhede plasmied DNS was opgegroei en die nukleotied volgorde bepaal op beide stringe van die genoom. Die stamme was gesubtipeer en filogenetiese analise was uitgevoer met standaard metodes. Die volledige DNS volgordes was bepaal en subtipering het daarop gedui dat die stamme van Tygerberg suiwer subtipe D is. Geen premature stop kodons is in die nukleotied volgordes van die Tygerberg stamme gevind nie. By die draai van die varieerbare deel (V3) het al die Tygerberg stamme die GQGQ motief gehad. R214 het 'n meer varieerbare vpu geen, maar behoort steeds tot die subtipe D groep in die gedeelte. Daar was geen premature stop kodons in die tat geen gevind nie en die glikosilasie van die stamme is minder as die van die konsensus subtipe D stam. Ons is die eerste groep om vollengte subtipe D stamme van Suid-Afrika te karakteriseer. Die DNS volgordes verteenwoordig suiwer subtipe D genome. Ons resultate bevestig die van ander dat die nukleotied volgordes van die ouer subtipe D stamme verskil van die nuwer stamme. 2010-06-29T13:04:29Z 2010-06-29T13:04:29Z 2004-12 Thesis http://hdl.handle.net/10019.1/3266 en application/pdf
spellingShingle HIV-1
Homosexual transmission
Virusses
Epidemics
DNA
Cloning
Glycosilation
Evolutionary phylogenetic trees
Tygerberg HIV-1 subtype D strains
HIV infections -- Transmission -- Prevention -- South Africa
DNA viruses -- South Africa
Loxton, Andre Gareth
Janse van Rensburg, E.
Engelbrecht, S.
Characterisation of new full-length HIV-1 subtype D viruses from South Africa
title Characterisation of new full-length HIV-1 subtype D viruses from South Africa
title_full Characterisation of new full-length HIV-1 subtype D viruses from South Africa
title_fullStr Characterisation of new full-length HIV-1 subtype D viruses from South Africa
title_full_unstemmed Characterisation of new full-length HIV-1 subtype D viruses from South Africa
title_short Characterisation of new full-length HIV-1 subtype D viruses from South Africa
title_sort characterisation of new full length hiv 1 subtype d viruses from south africa
topic HIV-1
Homosexual transmission
Virusses
Epidemics
DNA
Cloning
Glycosilation
Evolutionary phylogenetic trees
Tygerberg HIV-1 subtype D strains
HIV infections -- Transmission -- Prevention -- South Africa
DNA viruses -- South Africa
url http://hdl.handle.net/10019.1/3266
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