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目的定性定量地考察铜绿假单胞菌噬菌体PaP3在感染宿主菌PA3后对宿主转录组的全局性调控作用。方法利用高通量链特异性的RNA-seq对PaP3感染宿主菌PA3后5个时间点(5 min、10 min、20 min、30 min、80 min)的转录本进行深度测序,并以铜绿假单胞菌PAO1株及噬菌体PaP3基因组序列为参照,通过生物信息学对检测数据做定性定量的分析。结果以未感染噬菌体的细菌为对照,在铜绿假单胞菌PA3被噬菌体PaP3感染后的不同时间点共检测到5 536个差异表达基因,共归属于27条PseudoCAP功能注释,KEGG Pathway显著性富集分析发现差异表达基因共涉及45条代谢路径。此外,在5个样本中还预测出1 438个新转录本及1 329个新的候选sRNA,发现19种已注释的sRNA及91种新的候选sRNA出现差异表达。结论噬菌体PaP3可能通过抑制宿主菌PA3转录调节相关基因而对宿主的转录组进行全局性的调控,这为深入理解噬菌体与宿主相互作用的机制奠定了基础,也为探索噬菌体通过细菌与人体免疫系统的相互作用提供了多方面多层次的信息。
Objective To qualitatively and quantitatively investigate the global regulatory effect of PaP3, a Pseudomonas aeruginosa bacteriophage, on the host transcriptome after infection of host strain PA3. Methods Transcripts of PaP3-infected host PA3 at 5 time points (5 min, 10 min, 20 min, 30 min, 80 min) were sequenced by high-throughput RNA-seq. PAO1 strain and bacteriophage PaP3 genome sequences as a reference, the qualitative and quantitative analysis of the test data by bioinformatics. The results of non-infected bacteriophage bacteria as a control, Pseudomonas aeruginosa PA3 bacteriophage PaP3 infection at different times after the detection of a total of 5 536 differentially expressed genes, a total of 27 PseudoCAP functional annotation, KEGG Pathway significantly rich Set analysis found that differentially expressed genes involved in 45 metabolic pathways. In addition, 1 438 new transcripts and 1 329 new candidate sRNAs were also predicted in 5 samples, and 19 annotated sRNAs and 91 new candidate sRNAs were found to be differentially expressed. Conclusion The phage PaP3 may regulate the transcriptome of the host by inhibiting the transcriptional regulation genes of PA3, which lays a foundation for understanding the mechanism of the interaction between phage and host. It also aims to explore the mechanism of bacteriophage through the bacterial and human immune system The interaction provides a multifaceted, multi-level message.