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通过深度测序鉴定感染结核分枝杆菌 H37Rv 和卡介苗的 RAW264.7 的不同转录谱。

Different transcriptional profiles of RAW264.7 infected with Mycobacterium tuberculosis H37Rv and BCG identified via deep sequencing.

机构信息

Laboratory of Food Safety and Hygienic Inspection, Jilin University, Changchun, Jilin, People's Republic of China.

出版信息

PLoS One. 2012;7(12):e51988. doi: 10.1371/journal.pone.0051988. Epub 2012 Dec 19.

Abstract

BACKGROUND

The Mycobacterium tuberculosis H37Rv and BCG effects on the host cell transcriptional profile consider a main research point. In the present study the transcriptome profiling analysis of RAW264.7 either infected with Mycobacterium tuberculosis H37Rv or BCG have been reported using Solexa/Illumina digital gene expression (DGE).

RESULTS

The DGE analysis showed 1,917 different expressed genes between the BCG and H37Rv group. In addition, approximately 5% of the transcripts appeared to be predicted genes that have never been described before. KEGG Orthology (KO) annotations showed more than 71% of these transcripts are possibly involved in approximately 210 known metabolic or signaling pathways. The gene of the 28 pathways about pathogen recognition receptors and Mycobacterium tuberculosis interaction with macrophages were analyzed using the CLUSTER 3.0 available, the Tree View tool and Gene Orthology (GO). Some genes were randomly selected to confirm their altered expression levels by quantitative real-time PCR (qRT-PCR).

CONCLUSION

The present study used DGE from pathogen recognition receptors and Mycobacterium tuberculosis interaction with macrophages to understand the interplay between Mycobacterium tuberculosis and RAW264.7. Meanwhile find some important host protein which was affected by Mycobacterium tuberculosis to provide evidence for the further improvement of the present efficacy of existing Mycobacterium tuberculosis therapy and vaccine.

摘要

背景

结核分枝杆菌 H37Rv 和卡介苗对宿主细胞转录谱的影响是主要研究点。本研究采用 Solexa/Illumina 数字基因表达(DGE)技术,对 RAW264.7 细胞感染结核分枝杆菌 H37Rv 或卡介苗后的转录组进行了分析。

结果

DGE 分析显示,BCG 组和 H37Rv 组之间有 1917 个差异表达基因。此外,大约 5%的转录本似乎是以前从未描述过的预测基因。KEGG 直系同源(KO)注释显示,这些转录本中的 71%以上可能参与了大约 210 个已知的代谢或信号通路。使用 CLUSTER 3.0 可用的、Tree View 工具和基因直系同源(GO)分析了与病原体识别受体和结核分枝杆菌与巨噬细胞相互作用相关的 28 条通路的基因。随机选择一些基因通过定量实时 PCR(qRT-PCR)来验证其表达水平的变化。

结论

本研究采用 DGE 从病原体识别受体和结核分枝杆菌与巨噬细胞的相互作用来了解结核分枝杆菌与 RAW264.7 之间的相互作用。同时发现一些受结核分枝杆菌影响的重要宿主蛋白,为进一步提高现有结核分枝杆菌治疗和疫苗的疗效提供依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ccd/3526534/03b4292c6e7a/pone.0051988.g001.jpg

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