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人类嗜T细胞病毒III型3'开放阅读框编码的蛋白质在细菌中的表达:其与人血清免疫反应性的证明

Expression of the protein encoded by the 3' open reading frame of human T-cell lymphotropic virus type III in bacteria: demonstration of its immunoreactivity with human sera.

作者信息

Franchini G, Robert-Guroff M, Wong-Staal F, Ghrayeb J, Kato I, Chang T W, Chang N T

出版信息

Proc Natl Acad Sci U S A. 1986 Jul;83(14):5282-5. doi: 10.1073/pnas.83.14.5282.

Abstract

The genome of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV), the infectious agent etiologically associated with the acquired immunodeficiency syndrome, contains, in addition to the genes for the polymerase, core, and envelope proteins, several open reading frames. To investigate whether the 3' open reading frame (3' orf) located between the envelope gene and the 3' long terminal repeat is a gene expressed in vivo in infected individuals, we inserted a fragment of 3' orf in a prokaryotic expression vector. The protein product synthesized in bacteria was purified and allowed to react with sera from individuals infected with human T-cell lymphotropic virus type III as indicated by seropositivity for other viral proteins. Two-thirds of the sera, regardless of the clinical status of the individuals, reacted with the purified protein indicating that 3' orf is a viral gene the product of which is immunogenic in vivo. A polyclonal rabbit antiserum reacting against the 3' orf gene product was obtained by serial injection of rabbits with the purified bacterial protein. The antiserum recognized a 27-kDa protein in the human T-cell lymphotropic virus type III-infected lymphocytes.

摘要

人类嗜T细胞病毒III型/淋巴结病相关病毒(HTLV-III/LAV)是与获得性免疫缺陷综合征病因相关的感染因子,其基因组除了包含聚合酶、核心蛋白和包膜蛋白的基因外,还含有几个开放阅读框。为了研究位于包膜基因和3'长末端重复序列之间的3'开放阅读框(3'orf)是否为感染个体体内表达的基因,我们将3'orf片段插入原核表达载体中。在细菌中合成的蛋白质产物被纯化,并使其与感染人类嗜T细胞病毒III型个体的血清反应,这些个体对其他病毒蛋白的血清学检测呈阳性。无论个体的临床状态如何,三分之二的血清与纯化蛋白发生反应,表明3'orf是一种病毒基因,其产物在体内具有免疫原性。通过用纯化的细菌蛋白连续注射兔子,获得了针对3'orf基因产物的多克隆兔抗血清。该抗血清在感染人类嗜T细胞病毒III型的淋巴细胞中识别出一种27 kDa的蛋白质。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f640/323935/032a1e1d5c4d/pnas00318-0318-a.jpg

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