Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles.
dc.contributor.author | Halsey, Richard J | |
dc.contributor.author | Tanzer, Fiona L | |
dc.contributor.author | Meyers, Ann | |
dc.contributor.author | Pillay, Sirika | |
dc.contributor.author | Lynch, Alisson | |
dc.contributor.author | Shephard, Enid | |
dc.contributor.author | Williamson, Anna-Lise | |
dc.contributor.author | Rybicki, Edward P | |
dc.date.accessioned | 2016-07-20T13:54:51Z | |
dc.date.available | 2016-07-20T13:54:51Z | |
dc.date.issued | 2008 | |
dc.date.updated | 2016-07-20T13:53:05Z | |
dc.description.abstract | HIV-1 Pr55 Gag virus-like particles (VLPs) are strong immunogens with potential as candidate HIV vaccines. VLP immunogenicity can be broadened by making chimaeric Gag molecules: however, VLPs incorporating polypeptides longer than 200 aa fused in frame with Gag have not yet been reported. We constructed a range of gag-derived genes encoding in-frame C-terminal fusions of myristoylation-competent native Pr55Gag and p6-truncated Gag (Pr50Gag) to test the effects of polypeptide length and sequence on VLP formation and morphology, in an insect cell expression system. Fused sequences included a modified reverse transcriptase-Tat-Nef fusion polypeptide (RTTN, 778 aa), and truncated versions of RTTN ranging from 113 aa to 450 aa. Baculovirus-expressed chimaeric proteins were examined by western blot and electron microscopy. All chimaeras formed VLPs which could be purified by sucrose gradient centrifugation. VLP diameter increased with protein MW, from ∼100 nm for Pr55Gag to ∼250 nm for GagRTTN. The presence or absence of the Gag p6 region did not obviously affect VLP formation or appearance. GagRT chimaeric particles were successfully used in mice to boost T-cell responses to Gag and RT that were elicited by a DNA vaccine encoding a GagRTTN polypeptide, indicating the potential of such chimaeras to be used as candidate HIV vaccines. | en_ZA |
dc.identifier | http://dx.doi.org/10.1016/j.virusres.2008.01.012 | |
dc.identifier.apacitation | Halsey, R. J., Tanzer, F. L., Meyers, A., Pillay, S., Lynch, A., Shephard, E., ... Rybicki, E. P. (2008). Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles. <i>Virus Research</i>, http://hdl.handle.net/11427/20553 | en_ZA |
dc.identifier.chicagocitation | Halsey, Richard J, Fiona L Tanzer, Ann Meyers, Sirika Pillay, Alisson Lynch, Enid Shephard, Anna-Lise Williamson, and Edward P Rybicki "Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles." <i>Virus Research</i> (2008) http://hdl.handle.net/11427/20553 | en_ZA |
dc.identifier.citation | de Villiers, K. A., Marques, H. M., & Egan, T. J. (2008). The crystal structure of halofantrine–ferriprotoporphyrin IX and the mechanism of action of arylmethanol antimalarials. Journal of inorganic biochemistry, 102(8), 1660-1667. | en_ZA |
dc.identifier.issn | 0168-1702 | en_ZA |
dc.identifier.ris | TY - Journal Article AU - Halsey, Richard J AU - Tanzer, Fiona L AU - Meyers, Ann AU - Pillay, Sirika AU - Lynch, Alisson AU - Shephard, Enid AU - Williamson, Anna-Lise AU - Rybicki, Edward P AB - HIV-1 Pr55 Gag virus-like particles (VLPs) are strong immunogens with potential as candidate HIV vaccines. VLP immunogenicity can be broadened by making chimaeric Gag molecules: however, VLPs incorporating polypeptides longer than 200 aa fused in frame with Gag have not yet been reported. We constructed a range of gag-derived genes encoding in-frame C-terminal fusions of myristoylation-competent native Pr55Gag and p6-truncated Gag (Pr50Gag) to test the effects of polypeptide length and sequence on VLP formation and morphology, in an insect cell expression system. Fused sequences included a modified reverse transcriptase-Tat-Nef fusion polypeptide (RTTN, 778 aa), and truncated versions of RTTN ranging from 113 aa to 450 aa. Baculovirus-expressed chimaeric proteins were examined by western blot and electron microscopy. All chimaeras formed VLPs which could be purified by sucrose gradient centrifugation. VLP diameter increased with protein MW, from ∼100 nm for Pr55Gag to ∼250 nm for GagRTTN. The presence or absence of the Gag p6 region did not obviously affect VLP formation or appearance. GagRT chimaeric particles were successfully used in mice to boost T-cell responses to Gag and RT that were elicited by a DNA vaccine encoding a GagRTTN polypeptide, indicating the potential of such chimaeras to be used as candidate HIV vaccines. DA - 2008 DB - OpenUCT DP - University of Cape Town J1 - Virus Research LK - https://open.uct.ac.za PB - University of Cape Town PY - 2008 SM - 0168-1702 T1 - Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles TI - Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles UR - http://hdl.handle.net/11427/20553 ER - | en_ZA |
dc.identifier.uri | http://hdl.handle.net/11427/20553 | |
dc.identifier.uri | http://www.sciencedirect.com/science/article/pii/S0168170208000531 | |
dc.identifier.vancouvercitation | Halsey RJ, Tanzer FL, Meyers A, Pillay S, Lynch A, Shephard E, et al. Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles. Virus Research. 2008; http://hdl.handle.net/11427/20553. | en_ZA |
dc.language | eng | en_ZA |
dc.publisher | Elsevier | en_ZA |
dc.publisher.institution | University of Cape Town | |
dc.source | Virus Research | en_ZA |
dc.source.uri | http://www.sciencedirect.com/science/article/pii/S0162013408001141 | |
dc.subject.other | HIV | |
dc.subject.other | HIV vaccine | |
dc.subject.other | Virus-like particles | |
dc.subject.other | Chimaeric protein | |
dc.subject.other | Gag | |
dc.title | Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles. | en_ZA |
dc.type | Journal Article | en_ZA |
uct.type.filetype | Text | |
uct.type.filetype | Image | |
uct.type.publication | Research | en_ZA |
uct.type.resource | Article | en_ZA |