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目的明确铜绿假单胞菌噬菌体PaP3在感染宿主菌PA3后其全基因组的分时期表达模式。方法利用基因芯片检测噬菌体PaP3在感染宿主菌PA3后不同时间(5 min,10 min,20 min,30 min,80 min)其全基因组(含71个预测的ORF)在转录水平的表达谱变化,并进行非监督层次聚类(hierarchical clustering)分析。利用蛋白质合成抑制剂氯霉素(Cm)和DNA复制抑制剂磷酸乙酸(PAA)实验确定PaP3全基因组的分时期表达情况。结果根据时间表达模式的不同,PaP3全基因组可分为3大类:15个早期基因(ORF 71-57)、35个中期基因(ORF 56-22)及21个晚期基因(ORF 21-1)。在PaP3基因组结构中,这3类基因各自串联分布且可能受不同的操纵子调控。结论 PaP3感染过程中其早期、中期及晚期基因随时间有序表达,为深入了解噬菌体基因表达模式及生命复制周期奠定基础,并为了解噬菌体与宿主及机体免疫系统的的相互作用提供了基础。
Objective To determine the temporal pattern of genome-wide expression of Pseudomonas aeruginosa bacteriophage PaP3 after infection with PA3. Methods The expression profile of the whole genome (including 71 predicted ORFs) of the phage PaP3 phage at the transcriptional level was detected by gene chip at different times (5 min, 10 min, 20 min, 30 min, 80 min) And conducts unsupervised hierarchical clustering analysis. Epidermal expression of PaP3 whole genome was determined by using the protein synthesis inhibitor chloramphenicol (Cm) and DNA replication inhibitor phosphoacetate (PAA). Results According to the temporal expression pattern, PaP3 genome could be divided into three major groups: 15 early genes (ORF 71-57), 35 metaphase genes (ORF 56-22) and 21 late genes (ORF 21-1) . In the PaP3 genome structure, these three types of genes are each distributed in tandem and may be regulated by different operons. Conclusions The genes of early, middle and late stages of PaP3 infection are orderly expressed with time in order to understand the pattern of phage gene expression and life cycle replication and provide the basis for understanding the interaction between phage and host and the immune system.