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Construction and characterization of a full-length infectious clone from a fast-replicating, X4-tropic HIV-1 subtype B′ isolate

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Abstract

In HIV-1 epidemics in China, HIV-1 subtype B′ is the most predominant subtype circulating in intravenous drug users. In this study, we constructed an HIV-1 full-length infectious molecular clone based on the primary virus LWJ, which was isolated from an HIV-infected patient in Fujian Province, China. Phylogenetic and bootscanning analysis of the viral sequence revealed that the isolate LWJ belonged to HIV-1 subtype B′. The infectious clone was designated as “pLWJ”. The virus (LWJ-c) produced from this infectious clone by in vitro transfection of 293T cells could infect both human peripheral blood mononuclear cells (PBMCs) and human the T cell line MT4. Interestingly, the cloned LWJ-c virus utilized CXCR4 as its co-receptor and could replicate in vitro with similar efficiency and kinetics compared to its parental primary isolate LWJ as well as the clade B reference virus NL4-3. The LWJ-c virus could also cause cytopathic effects in both PBMCs and MT cells. Sequence analysis of the envelope glycoprotein of pLWJ showed that a conserved GPGR motif and an arginine at position 11 were present in the V3 loop, which was consistent with previous reports regarding CXCR4 co-receptor usage and syncytium-inducing (SI) phenotype. Thus, the infectious clone represents a fast-replicating, high-producing, CXCR4-tropic and syncytium-inducing isolate. Given the prevalence of HIV-1 subtype B′ in China, this infectious clone can be a very useful tool to provide a versatile molecular model for research focusing on the biological properties of this subtype.

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Acknowledgments

This work was supported by a grant from Natural Science Foundation of Fujian Province (No. 2009J01104). We are grateful to Professor Shixia Wang from the University of Massachusetts Medical School for her critical editing of the manuscript.

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Authors and Affiliations

  1. Fujian Medical University, Fuzhou, Fujian, China

    Shou-li Wu & Yan-sheng Yan

  2. Fujian Center for Disease Control and Prevention (FJCDC), 76 Jin Tai Lu, Fuzhou, 350001, Fujian, People’s Republic of China

    Shou-li Wu, Yan-sheng Yan, Ping-ping Yan & Hui-rong Wang

  3. Maternal and Child Health Care Hospital of Fujian Province, Fuzhou, Fujian, China

    Hai-long Huang

Authors
  1. Shou-li Wu
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  2. Yan-sheng Yan
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  3. Ping-ping Yan
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  4. Hai-long Huang
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Corresponding author

Correspondence to Yan-sheng Yan.

Appendix

Appendix

The complete sequences of the molecular clones pLWJ10 + 6, pLWJ11 + 6, pLWJ5 + 6, pLWJ2 + 6 were deposited in GenBank with accession numbers GU177863, GU647196, GU647197, GU647198, respectively.

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Wu, Sl., Yan, Ys., Yan, Pp. et al. Construction and characterization of a full-length infectious clone from a fast-replicating, X4-tropic HIV-1 subtype B′ isolate. Arch Virol 155, 1923–1931 (2010). https://doi.org/10.1007/s00705-010-0776-z

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  • DOI: https://doi.org/10.1007/s00705-010-0776-z

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