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Construction and Characterization of Human Immunodeficiency Virus Type 1 Subtype C Clones with env Genes from Divergent Phylogenetic Lineages M Modukanele*, K Armstrong, T Ndung’u, VA Novitsky, E Sepako, and M Essex Department of Immunology and Infectious Diseases, Harvard University, Boston, MA, USA Background: Human immunodeficiency virus type 1 subtype C is now responsible for the majority of infections in the HIV/AIDS pandemic and is the predominant virus in Southern Africa. Although the reasons for the rapid spread of HIV-1 subtype C remain largely unclear, HIV-1 subtype C possesses unique genetic features that may result in important functional consequences. Methods: We identified HIV-1 subtype C envelope genes that lack obvious inactivating mutations from representative near full-length HIV-1 subtype C from Botswana from different phylogenetic lineages. We then used standard molecular cloning techniques to generate full-length HIV-1 subtype C clones expressing the different env clones, all in the backbone of the previously generated infectious molecular clone pMJ4. Results: We have generated five full-length HIV-1 subtype C clones possessing genetically divergent env genes. Preliminary results from transient transfection studies followed by infection of HIV-negative donor peripheral blood mononuclear cells suggest that some of these clones are infectious. Further studies on growth and replication kinetics in donor PBMCs and susceptible cell lines are in progress. Co-receptor utilization by these clones will also be characterized. Conclusions/Summary: We have generated a panel of HIV-1 subtype C clones that may facilitate studies of mechanisms of disease pathogenesis and vaccine development for HIV-1 subtype C. |