Construction and Characterization of a Full-Length Infectious Molecular Clone from the HIV Type 1 Subtype Thai-B Isolated in Henan Province, China

Wang, Zheng; Li, Jinyun; Li, Lin; Feng, Fuming; Li, Hanping; Bao, Zhenmin

doi:10.1089/aid.2007.0226

Cited by 7 publications

(4 citation statements)

References 15 publications

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“…To date, almost all HIV infectious clones were constructed using very high-copy number vectors such as those from the pUC series [16], [21], [22], [39]. In this present study, the first molecular clone pNGX5, comprising the pNL4-3 vector and the 2007CNGX-HK genome, except for its LTRs, was constructed using a similar strategy.…”

Section: Discussionmentioning

confidence: 99%

Parental LTRs Are Important in a Construct of a Stable and Efficient Replication-Competent Infectious Molecular Clone of HIV-1 CRF08_BC

Zhang

et al. 2012

PLoS ONE

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Circulating recombinant forms (CRFs) of HIV-1 have been identified in southern China in recent years. CRF08_BC is one of the most predominant subtypes circulating in China. In order to study HIV subtype biology and to provide a tool for biotechnological applications, the first full-length replication-competent infectious molecular clone harboring CRF08_BC is reported. The construction of this clone pBRGX indicates that a moderate-copy number vector is required for its amplification in E. coli. In addition, it is shown that the parental CRF08_BC LTRs are important for generating this efficient replication-competent infectious clone. These observations may aid in the construction of infectious clones from other subtypes. Both the pBRGX-derived virus and its parental isolate contain CCR5 tropism. Their full-length genomes were also sequenced, analyzed, compared and deposited in GenBank (JF719819 and JF719818, respectively). The availability of pBRGX as the first replication-competent molecular clone of CRF08_BC provides a useful tool for a wide range of studies of this newly emergent HIV subtype, including the development of HIV vaccine candidates, antiviral drug screening and drug resistance analysis.

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Section: Discussionmentioning

confidence: 99%

Parental LTRs Are Important in a Construct of a Stable and Efficient Replication-Competent Infectious Molecular Clone of HIV-1 CRF08_BC

Zhang

et al. 2012

PLoS ONE

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“…Specific primers were edited to fulfill the seminested PCR to obtain the full-length HIV DNA provirus (Table 1). 7,8 The two partial molecules of the entire S15 genome were amplified with the Platinum Taq DNA polymerase High Fidelity kit (Invitrogen, Carlsberg, CA).…”

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confidence: 99%

Genetic Characterization of a Unique Recombinant Originating from CRF55_01B, CRF01_AE, and CRF07_BC in Shenzhen, China

Gui

Zhao

Sun

et al. 2015

AIDS Research and Human Retroviruses

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Multiple subtypes were found to be epidemic in the Shenzhen men who have sex with men (MSM) population, which always predicts the emergence of a unique recombinant. In 2012, CRF55_01B was first reported, which later was proven to have originated in MSM in Shenzhen city. In this study, we reported a unique recombinant form (URF) of HIV-1 identified in a man who has had sex with men in Shenzhen city. The strain showed a genomic schematic map similar to CRF55_01B with subtype C segments inserted in the gag and pol genes. The full-length genome was amplified in two halves with 1-kb overlap regions. The PCR products were cloned and sequenced. A recombination detection program showed that two subtype C fragments and two subtype B fragments were inserted into the CRF01_AE backbone genome in the gag and pol regions. In the phylogenetic tree, the subtype C fragments clustered with CRF07_BC variants and the other segments grouped with CRF55_01B strains except for one segment that clustered with CRF01_AE. Similar breakpoints between our strain and CRF65_cpx were also observed. The data suggested that the URF strain might be the recombinant form of CRF55_01B, CRF01_AE, and CRF07_BC. This is the first report of a third generation of recombination of HIV-1 that originated from CRF55_01B in China. The identification of the URF indicated the severity of the HIV epidemic in Shenzhen MSM and the urgent need for epidemiological surveillance of the new recombination.

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“…Phenotypic and genotypic studies that involve artificial manipulation of viral genomes at DNA level provide significant insights on viral replication, pathogeneses and other biological characteristics. Despite being one of the major sites for CRF01_AE and subtype B-based vaccine efficacy trials, the limited collection of molecular clones of Southeast Asian origin [13], [20]–[22] may present a daunting challenge in developing vaccine candidates that match the locally prevalent and diverse HIV strains especially when there is a lack of standardized reagents for HIV neutralization assays in clinical evaluations. As an emerging, widely distributed CRF in the Southeast Asia region, an CRF33_01B infectious DNA clone is a valuable tool in facilitating the development of broadly effective vaccine candidates.…”

Section: Resultsmentioning

confidence: 99%

Isolation and Characterization of a Replication-Competent Molecular Clone of an HIV-1 Circulating Recombinant Form (CRF33_01B)

Tee

Kusagawa

et al. 2009

PLoS ONE

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A growing number of emerging HIV-1 recombinants classified as circulating recombinant forms (CRFs) have been identified in Southeast Asia in recent years, establishing a molecular diversity of increasing complexity in the region. Here, we constructed a replication-competent HIV-1 clone for CRF33_01B (designated p05MYKL045.1), a newly identified recombinant comprised of CRF01_AE and subtype B. p05MYKL045.1 was reconstituted by cloning of the near full-length HIV-1 sequence from a newly-diagnosed individual presumably infected heterosexually in Kuala Lumpur, Malaysia. The chimeric clone, which contains the 5′ LTR (long terminal repeat) region of p93JP-NH1 (a previously isolated CRF01_AE infectious clone), showed robust viral replication in the human peripheral blood mononuclear cells. This clone demonstrated robust viral propagation and profound syncytium formation in CD4+, CXCR4-expressing human glioma NP-2 cells, indicating that p05MYKL045.1 is a CXCR4-using virus. Viral propagation, however, was not detected in various human T cell lines including MT-2, M8166, Sup-T1, H9, Jurkat, Molt-4 and PM1. p05MYKL045.1 appears to proliferate only in restricted host range, suggesting that unknown viral and/or cellular host factors may play a role in viral infectivity and replication in human T cell lines. Availability of a CRF33_01B molecular clone will be useful in facilitating the development of vaccine candidates that match the HIV-1 strains circulating in Southeast Asia.

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Construction and Characterization of a Full-Length Infectious Molecular Clone from the HIV Type 1 Subtype Thai-B Isolated in Henan Province, China

Cited by 7 publications

References 15 publications

Parental LTRs Are Important in a Construct of a Stable and Efficient Replication-Competent Infectious Molecular Clone of HIV-1 CRF08_BC

Parental LTRs Are Important in a Construct of a Stable and Efficient Replication-Competent Infectious Molecular Clone of HIV-1 CRF08_BC

Genetic Characterization of a Unique Recombinant Originating from CRF55_01B, CRF01_AE, and CRF07_BC in Shenzhen, China

Isolation and Characterization of a Replication-Competent Molecular Clone of an HIV-1 Circulating Recombinant Form (CRF33_01B)

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