感染多耐药大肠杆菌新噬菌体vB_EcoM_RZ的性质鉴定及基因组学研究
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国家自然科学基金(81501797)


Characterization and genomic study of phage vB_EcoM_RZ infecting multidrug⁃resistant Escherichia coli
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    摘要:

    目的:分离鉴定新型毒性噬菌体并研究其生物学特性,为用于治疗临床多耐药菌感染提供备选方案。方法:从南京城区污水中收集废水样本,以临床分离的大肠杆菌多药耐药菌株为宿主,采用双层琼脂法分离纯化新噬菌体,然后采用电镜观察其形态,一步生长曲线确定其潜伏期及爆发量,并通过全基因组测序及质谱分析结合生物信息学分析研究其基因组及编码蛋白。结果:成功分离出一种新型大肠杆菌毒性噬菌体命名为vB_EcoM_RZ,其能感染并裂解多株大肠杆菌临床耐药菌株;生物学性质分析表明该噬菌体裂解速度快、宿主范围广、温度稳定性良好;全基因组测序结果表明vB_EcoM_RZ不编码毒素或其他毒力因子基因,溶源周期相关基因及抗性基因;质谱分析发现vB_EcoM_RZ噬菌体颗粒含有43种已知蛋白和一些未知功能的假定蛋白;系统发育树分析表明vB_EcoM_RZ是一株新型噬菌体。结论:分离并鉴定出一株新型大肠杆菌毒性噬菌体vB_EcoM_RZ,性质优良,有进一步应用于治疗耐药菌引起感染的潜能。

    Abstract:

    Objective:Isolate and identify a new strain of lytic phage and study its biological characteristics,so as to provide an alternative treatment for clinical infection caused by multi-drug resistant bacterial in the future. Methods:The new phage named vB_EcoM_RZ was isolated and purified by double layer agar method. The morphology of the new phage was observed by electron microscope. The incubation period and burst size of the new phage were determined by one-step growth curve,the genomic characteristics and encoded proteins of phage vB_EcoM_RZ were studied through the whole genome sequencing and mass spectrometry analysis combined with bioinformatics analysis. Results:A new type of Escherichia coli virulent bacteriophage was successfully isolated;biological experiments showed that it had fast lytic speed,wide host range,and good temperature stability;genomics studies showed that it did not encode genes related to toxins or other virulence factors;43 known proteins and some hypothetical proteins with unknown functions were isolated and identified by mass spectrometry. Phylogenetic analysis showed that it was a new type of bacteriophage. Conclusion:A new type of Escherichia coli virulent bacteriophage named vB_EcoM_RZ has been isolated and identified. It has excellent properties and the potential to be further used in the treatment of infection caused by drug-resistant bacteria.

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张瑞阳,许 靖,于鑫焱,刘晓秋.感染多耐药大肠杆菌新噬菌体vB_EcoM_RZ的性质鉴定及基因组学研究[J].南京医科大学学报(自然科学版),2021,(8):1109-1114

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  • 收稿日期:2021-04-25
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  • 在线发布日期: 2021-08-30
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