MSMEG_6171在耻垢分枝杆菌中的表达及多组学关联分析

Expression and multi-omics analysis of MSMEG_6171 in Mycobacterium smegmatis

目的筛选MSMEG_6171在耻垢分枝杆菌中的可能调控通路或基因,并进行验证。方法构建MSMEG_6171基因过表达的耻垢分枝杆菌株和空载质粒对照菌株;对其转录组和蛋白质组进行验证并深入挖掘MSMEG_6171过表达调控的下游信号网络;qRT-PCR验证差异表达基因。结果成功构建了MSMGE_6171过表达的耻垢分枝杆菌菌株(Msm::6171);转录组和蛋白质组数据的对比分析发现有38个基因在转录水平和蛋白表达水平显著上调,有31个基因表达水平显著下调;差异表达基因主要参与甘油酯代谢,核糖体和代谢通路;15个候选基因实时荧光定量PCR验证结果与转录组和蛋白质组测序分析趋势一致。结论MSMGE_6171调控了多个靶向基因,参与代谢和核糖体通路,为深入研究MSMGE_6171的调控网络提供了新的认识和重要参考。

This study screened the potential genes or pathways regulated by MSMEG_6171 in Mycobacterium smegmatis and to provide a preliminary verification of the results.We constructed a recombinant MSMEG_6171 overexpression strain and control plasmid strain by using molecular cloning techniques.Second-generation high-throughput sequencing technology and iTRAQ-based proteomics analysis were used to compare the transcriptomic and proteomic data.GO and KEGG pathway enrichment analyses were used to analyze gene function.Then differentially expressed genes(DEGs)were verified by real-time RT-PCR(qRT-PCR).A recombinant MSMEG_6171 overexpression strain(Msm::6171)was constructed.A total of 69 DEGs in the MSMEG_6171 overexpression strain relative to the WT strain were identified,including 38 up-regulated genes and 31 down-regulated genes.GO enrichment analysis indicated that these DEGs were associated with functions including glycerol catabolic process,glycerol-3-phosphate metabolic process,translation,ribosome subunit and enzyme activity.KEGG pathway enrichment analysis demonstrated that the DEGs were involved in glycerolipid metabolism,ribosomes,and metabolic pathways.The qRT-PCR results for 15 candidate genes showed the same trends at the transcriptomicand proteomic levels.The results uncovered genes and signaling networks regulated by MSMEG_6171,thus providing a new understanding and serving as a reference for further study of the regulation network of MSMEG_6171.