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首页> 外文期刊>The FEBS journal >Construction and biological activity of a full-length molecular clone of human Torque teno virus (TTV) genotype 6
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Construction and biological activity of a full-length molecular clone of human Torque teno virus (TTV) genotype 6

机译：人T型转基因病毒（TTV）基因型6的全长分子克隆的构建及生物学活性

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摘要

Torque teno virus (TTV) is a non-enveloped human virus with a circular negative-sense (approximately 3800 nucleotides) ssDNA genome. TTV resembles in genome organization the chicken anemia virus, the animal pathogen of the Circoviridae family, and is currently classified as a member of a new, floating genus, Anellovirus. Molecular and cell biological research on TTV has been restricted by the lack of permissive cell lines and functional, replication-competent plasmid clones. In order to examine the key biological activities (i.e. RNA transcription and DNA replication) of this still poorly characterized ssDNA virus, we cloned the full-length genome of TTV genotype 6 and transfected it into cells of several types. TTV mRNA transcription was detected by RT-PCR in all the cell types: KU812Ep6, Cos-1, 293, 293T, Chang liver, Huh7 and UT7/Epo-S1. Replicating TTV DNA was detected in the latter five cell types by a DpnI-based restriction enzyme method coupled with Southern analysis, a novel approach to assess TTV DNA replication. The replicating full-length clone, the cell lines found to support TTV replication, and the methods presented here will facilitate the elucidation of the molecular biology and the life cycle of this recently identified human virus.
机译：扭力腱病毒（TTV）是一种非包膜人类病毒，具有环状负义（约3800个核苷酸）的ssDNA基因组。 TTV在基因组结构上类似于鸡贫血病毒（Circoviridae家族的动物病原体），目前被归类为新的浮动属Anellovirus的成员。 TTV的分子和细胞生物学研究因缺乏允许的细胞系和功能强大，具有复制能力的质粒克隆而受到限制。为了检查这种仍然特性欠佳的ssDNA病毒的关键生物学活性（即RNA转录和DNA复制），我们克隆了TTV基因型6的全长基因组并将其转染到几种类型的细胞中。通过RT-PCR在所有细胞类型中检测TTV mRNA转录：KU812Ep6，Cos-1、293​​、293T，Chang肝，Huh7和UT7 / Epo-S1。通过基于DpnI的限制性酶方法结合Southern分析（一种评估TTV DNA复制的新方法），在后五种细胞类型中检测到TTV DNA复制。复制的全长克隆，支持TTV复制的细胞系以及此处介绍的方法将有助于阐明这种新近鉴定的人类病毒的分子生物学和生命周期。

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