近期,，中科院武漢病毒所石正麗研究員領導的科研團隊采用第二代高通量測序技術,，對蝙蝠腺病毒BtAdV-TJM感染蝙蝠細胞系的轉錄譜做了系統(tǒng)的分析,，提供了一套完整的蝙蝠腺病毒復制過程中病毒基因表達的動態(tài)分布,。結果發(fā)表在2013年第1期Journal of Virology上,。
腺病毒廣泛存在于脊椎動物包括人和非人類靈長動物,，能引起人呼吸道和腸道疾病,。由于其良好的免疫原性,，腺病毒被開發(fā)為疫苗和基因治療的載體。但由于人體已有腺病毒的感染限制了人類腺病毒作為基因載體的廣泛應用,。因此,，需要開發(fā)其它動物腺病毒作為新型載體。
該課題組前期用蝙蝠原代細胞分離到一株新型蝙蝠腺病毒（BtAdV-TJM）（Li et al., JVI, 2010）,。本研究利用蝙蝠腺病毒感染大衛(wèi)鼠耳蝠細胞系,，采用高通量測序技術對病毒及宿主細胞的轉錄組進行系統(tǒng)研究。結果表明：隨著感染時間的增加,，蝙蝠腺病毒在蝙蝠細胞中轉錄復制加快,，在感染后18h達到頂峰。和人腺病毒感染類似,，蝙蝠腺病毒復制分為明顯早,，中，晚期三個階段,。早期基因主要編碼和宿主相互作用的一些蛋白,，中期基因編碼主要編碼與自身復制相關的蛋白，晚期基因編碼病毒裝配和結構蛋白,。RT-PCR確認有7個蝙蝠腺病毒基因轉錄復制不同于人腺病毒,。這預示著，蝙蝠腺病毒的轉錄復制機制可能與人和其他哺乳動物來源的腺病毒不同,。同時,，該研究對病毒感染后宿主細胞上調及下調基因進行功能分類，主要分為細胞免疫反應,、轉錄,、翻譯、細胞復制及修復等,，其中有79個宿主免疫基因在病毒感染后發(fā)生明顯上調,。上述研究結果為蝙蝠腺病毒載體的開發(fā)和蝙蝠免疫系統(tǒng)抗病毒免疫反應的研究提供了基礎。(生物谷Bioon.com)
doi: 10.1128/JVI.02332-12
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Deep RNA Sequencing Reveals Complex Transcriptional Landscape of a Bat Adenovirus
Lijun Wu, Peng Zhou, Xingyi Ge, Lin-Fa Wang, Michelle L. Baker and Zhengli Shi
Bat adenoviruses are a group of recently identified adenoviruses (AdVs) which are highly prevalent in bats yet share low similarity to known AdVs from other species. In this study, deep RNA sequencing was used to analyze the transcriptome at five time points following the infection of a bat AdV in a kidney cell line derived from a myotis bat species. Evidence of AdV replication was observed with the proportion of viral RNAs ranging from 0.01% at 6 h to 1.3% at 18 h. Further analysis of viral temporal gene expression revealed three replication stages, the early-stage genes encoding mainly host interaction proteins, the intermediate-stage genes for the DNA replication and assembly proteins, and the late-stage genes for most structural proteins. Several bat AdV genes were expressed at stages that differed from those of their counterpart genes previously reported for human AdV type 2. In addition, single-base resolution splice sites of several genes and promoter regions of all 30 viral genes were fully determined. Simultaneously, the temporal cellular gene expression profiles were identified. The most overrepresented functional categories of the differentially expressed genes were related to cellular immune response, transcription, translation, and DNA replication and repair. Taken together, the deep RNA sequencing provided a global, transcriptional profile of the novel bat AdV and the virus-host interactions which will be useful for the understanding and investigation of AdV replication, pathogenesis, and specific virus-bat interactions in future research.
