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来自尼日利亚的一种新型HIV-1 A亚型毒株的基因组结构与核苷酸序列分析

Genomic structure and nucleotide sequence analysis of a new HIV type 1 subtype A strain from Nigeria.

作者信息

Howard T M, Rasheed S

机构信息

Laboratory of Viral Oncology, and AIDS Research, University of Southern California, School of Medicine, Los Angeles 90032-3626, USA.

出版信息

AIDS Res Hum Retroviruses. 1996 Oct 10;12(15):1413-25. doi: 10.1089/aid.1996.12.1413.

Abstract

We have isolated and characterized a new HIV-1 variant (HIV-1IbNg) from the peripheral blood mononuclear cells (PBMCs) of an inhabitant of Nigeria. This virus is highly cytopathic to PBMCs in culture, replicates in primary human T cells and macrophages/monocytes as well as in established human T cell and monocytic cell lines (i.e., it has a wide host range), and it does not induce syncytia in MT-2 cells.1 Because of these unusual infectivity parameters in vitro, we analyzed the genetic structure of the entire genome. Using cytoplasmic RNA from HIV-1IbNg-infected PBMCs, five overlapping DNA fragments were amplified by the reverse transcriptase-polymerase chain reaction (RT-PCR) technique and cloned into pBluescript II SK(+). DNA sequencing of these fragments indicated that the entire HIV-1IbNg genome consisted of 9201 nucleotides and phylogenetic analysis of its env gene sequence revealed that this virus clustered with HIV-1 strains belonging to clade "A". In this article we present several genetic features unique to this virus, including (1) the presence of a 16-bp insert within the primer-binding site, (2) a large Rev open reading frame, (3) a Rev-responsive element that is predicted to form a different secondary structure than described for clade "B" viruses, (4) the potential to encode a heavily glycosylated Env protein, and (5) a frameshift, resulting in a stop codon, in the tat gene. This represents the first detailed analysis of the genetic structure of an HIV-1 strain from Nigeria.

摘要

我们从一名尼日利亚居民的外周血单个核细胞(PBMC)中分离并鉴定出一种新的HIV-1变异株(HIV-1IbNg)。该病毒在培养的PBMC中具有高度细胞病变性,可在原代人T细胞和巨噬细胞/单核细胞以及已建立的人T细胞和单核细胞系中复制(即它具有广泛的宿主范围),并且在MT-2细胞中不诱导形成多核巨细胞。由于这些体外异常的感染性参数,我们分析了整个基因组的遗传结构。使用来自HIV-1IbNg感染的PBMC的细胞质RNA,通过逆转录酶-聚合酶链反应(RT-PCR)技术扩增出五个重叠的DNA片段,并克隆到pBluescript II SK(+)中。这些片段的DNA测序表明,整个HIV-1IbNg基因组由9201个核苷酸组成,对其env基因序列的系统发育分析显示,该病毒与属于“A”亚型的HIV-1毒株聚类。在本文中,我们展示了该病毒特有的几个遗传特征,包括:(1)引物结合位点内存在一个16bp的插入片段;(2)一个大的Rev开放阅读框;(3)一个Rev反应元件,预计其形成的二级结构与“B”亚型病毒不同;(4)编码高度糖基化Env蛋白的潜力;(5)tat基因中的一个移码突变,导致一个终止密码子。这代表了对来自尼日利亚的HIV-1毒株遗传结构的首次详细分析。

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