Publicação
Genotypic and phenotypic characterization of HIV-1 virus found early in infection
| Resumo: | In this study we raised full-length envelope sequences from HIV-1 infected patients in early phases of infection, up to three month of viral acquisition, and from their transmitting partners, who were in later stages of infection, to assess any genotypic signature(s) that would distinguish the envelope sequences that were being transmitted and correlate those signatures with viral co-receptor usage and with drugs and neutralization sensitivity. Briefly, we isolated RNA from patients plasma, retro-transcribed it into cDNA and amplified envelope. We used yeast gap repair to clone envelope sequences into our expression vectors. Vectors were then transformed into bacteria and isolated clones were subjected to PCR and sequenced. Despite small sample size, V3 loop and V1-V5 region length seem to be shorter in earlier viruses when compared to later viruses. No other fragments analyzed from envelope showed any trend to be shorter in earlier isolates; neither did full-length envelope, gp41 or gp120. Also, no statistically significant decrease in the number of potential N-glycosylation sites or on the sequences genetic distances was establish in earlier virus. |
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| Autores principais: | Freitas, Inês Trindade de, 1986- |
| Assunto: | Biologia molecular HIV Manipulação genética Teses de mestrado - 2010 |
| Ano: | 2010 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | In this study we raised full-length envelope sequences from HIV-1 infected patients in early phases of infection, up to three month of viral acquisition, and from their transmitting partners, who were in later stages of infection, to assess any genotypic signature(s) that would distinguish the envelope sequences that were being transmitted and correlate those signatures with viral co-receptor usage and with drugs and neutralization sensitivity. Briefly, we isolated RNA from patients plasma, retro-transcribed it into cDNA and amplified envelope. We used yeast gap repair to clone envelope sequences into our expression vectors. Vectors were then transformed into bacteria and isolated clones were subjected to PCR and sequenced. Despite small sample size, V3 loop and V1-V5 region length seem to be shorter in earlier viruses when compared to later viruses. No other fragments analyzed from envelope showed any trend to be shorter in earlier isolates; neither did full-length envelope, gp41 or gp120. Also, no statistically significant decrease in the number of potential N-glycosylation sites or on the sequences genetic distances was establish in earlier virus. |
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