Molecular cloning and biological characterization of full-length HIV-1 subtype C from Botswana
Molecular cloning and biological characterization of full-length HIV-1 subtype C from Botswana
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DOI：
10.1006/viro.2000.0583
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发表时间：
2000-12-20
期刊：
影响因子：
3.7
通讯作者：
Essex, M
中科院分区：
文献类型：
--
作者：
Ndung'u, T;Renjifo, B;Essex, M
目前，人类免疫缺陷病毒1型（艾滋病毒-I）C亚型占全球艾滋病毒-1感染总数的一半以上，也是南部非洲艾滋病毒-1高度流行的原因。为了促进这种病毒的生物学性质和潜在的分子决定因素的研究，我们构建了8个全长前病毒克隆从两个无症状和三个艾滋病患者感染HIV-1亚型C从博茨瓦纳。病毒裂解物的分析表明，Gag，Pol和Env结构蛋白存在于病毒体中。在四个克隆中，分析表明包膜糖蛋白加工效率低下。8个克隆的核苷酸序列分析没有发现任何结构、调控或辅助基因中的移码、缺失、过早截短或翻译终止密码子。C亚型克隆在供体外周血单核细胞（PBMC）、巨噬细胞、Jurkat（达特）细胞或U87.CD4.CCR 5细胞中均无复制能力。然而，两个克隆的感染可以通过与功能亚型C包膜克隆互补来挽救，导致PBMC、巨噬细胞和U87.CD4.CCR 5细胞的生产性感染。(C)北京大学出版社.
Human immunodeficiency virus type 1 (HIV-I) subtype C is now responsible for more than half of all HIV-1 infections in the global epidemic and for the high levels of HIV-1 prevalence in southern Africa. To facilitate studies of the biological nature and the underlying molecular determinants of this virus, we constructed eight full-length proviral clones from two asymptomatic and three AIDS patients infected with HIV-1 subtype C from Botswana. Analysis of viral lysates showed that Gag, Pol, and Env structural proteins were present in the virions. In four clones, the analysis suggested inefficient envelope glycoprotein processing. Nucleotide sequence analysis of the eight clones did not reveal frameshifts, deletions, premature truncations, or translational stop codons in any structural, regulatory, or accessory genes. None of the subtype C clones were replication competent in donor peripheral blood mononuclear cells (PBMCs), macrophages, Jurkat(tat) cells, or U87.CD4.CCR5 cells. However, infection by two clones could be rescued by complementation with a functional subtype C envelope clone, resulting in a productive infection of PBMCs, macrophages, and U87.CD4.CCR5 cells. (C) 2000 Academic Press.