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从模式昆虫病原线虫嗜菌异小杆线虫中鉴定候选感染基因。

Identification of candidate infection genes from the model entomopathogenic nematode Heterorhabditis bacteriophora.

作者信息

Vadnal Jonathan, Ratnappan Ramesh, Keaney Melissa, Kenney Eric, Eleftherianos Ioannis, O'Halloran Damien, Hawdon John M

机构信息

Department of Biological Sciences, George Washington University, Science and Engineering Hall, suite 6000, 800 22nd Street NW, Washington DC, 20052, USA.

Department of Microbiology Immunology and Tropical Medicine, George Washington University Medical Center, Washington DC, 20037, USA.

出版信息

BMC Genomics. 2017 Jan 3;18(1):8. doi: 10.1186/s12864-016-3468-6.

Abstract

BACKGROUND

Despite important progress in the field of innate immunity, our understanding of host immune responses to parasitic nematode infections lags behind that of responses to microbes. A limiting factor has been the obligate requirement for a vertebrate host which has hindered investigation of the parasitic nematode infective process. The nematode parasite Heterorhabditis bacteriophora offers great potential as a model to genetically dissect the process of infection. With its mutualistic Photorhabdus luminescens bacteria, H. bacteriophora invades multiple species of insects, which it kills and exploits as a food source for the development of several nematode generations. The ability to culture the life cycle of H. bacteriophora on plates growing the bacterial symbiont makes it a very exciting model of parasitic infection that can be used to unlock the molecular events occurring during infection of a host that are inaccessible using vertebrate hosts.

RESULTS

To profile the transcriptional response of an infective nematode during the early stage of infection, we performed next generation RNA sequencing on H. bacteriophora IJs incubated in Manduca sexta hemolymph plasma for 9 h. A subset of up-regulated and down-regulated genes were validated using qRT-PCR. Comparative analysis of the transcriptome with untreated controls found a number of differentially expressed genes (DEGs) which cover a number of different functional categories. A subset of DEGs is conserved across Clade V parasitic nematodes revealing an array of candidate parasitic genes.

CONCLUSIONS

Our analysis reveals transcriptional changes in the regulation of a large number of genes, most of which have not been shown previously to play a role in the process of infection. A significant proportion of these genes are unique to parasitic nematodes, suggesting the identification of a group of parasitism factors within nematodes. Future studies using these candidates may provide functional insight into the process of nematode parasitism and also the molecular evolution of parasitism within nematodes.

摘要

背景

尽管在固有免疫领域取得了重要进展,但我们对宿主针对寄生线虫感染的免疫反应的了解仍落后于对微生物反应的了解。一个限制因素是对脊椎动物宿主的绝对需求,这阻碍了对寄生线虫感染过程的研究。线虫寄生虫嗜菌异小杆线虫作为一种对感染过程进行基因剖析的模型具有巨大潜力。嗜菌异小杆线虫与其共生的发光光杆状菌一起,可侵入多种昆虫,将其杀死并作为食物来源,供几代线虫发育。能够在培养细菌共生体的平板上培养嗜菌异小杆线虫的生命周期,使其成为一种非常令人兴奋的寄生虫感染模型,可用于揭示在感染宿主过程中发生的分子事件,而这些事件是使用脊椎动物宿主无法获得的。

结果

为了描绘感染性线虫在感染早期的转录反应,我们对在烟草天蛾血淋巴血浆中孵育9小时的嗜菌异小杆线虫感染性幼虫进行了下一代RNA测序。使用qRT-PCR验证了一组上调和下调基因。将转录组与未处理的对照进行比较分析,发现了许多差异表达基因(DEG),它们涵盖了许多不同的功能类别。一组DEG在进化枝V寄生线虫中是保守的,揭示了一系列候选寄生基因。

结论

我们的分析揭示了大量基因调控中的转录变化,其中大多数基因以前未被证明在感染过程中起作用。这些基因中有很大一部分是寄生线虫所特有的,这表明在 nematodes 中鉴定出了一组寄生因子。未来使用这些候选基因的研究可能会提供对线虫寄生过程以及线虫内寄生现象分子进化的功能见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c359/5209865/254d4e5f894f/12864_2016_3468_Fig1_HTML.jpg

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