| 一种简便的拓宽噬菌体宿主谱方法的建立及其结果分析 |
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| 引用本文： | 周艳,薛金鑫,叶敏,张辉,包红朵,王冉.一种简便的拓宽噬菌体宿主谱方法的建立及其结果分析[J].中国动物传染病学报,2020(1):8-15. |
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| 作者姓名： | 周艳 薛金鑫 叶敏 张辉 包红朵 王冉 |
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| 作者单位： | 江苏省农业科学院农产品质量安全与营养研究所江苏省食品质量安全重点实验室省部共建国家重点实验室培育基地;南京大学金陵学院化学与生命科学学院 |
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| 基金项目： | 国家自然科学基金(31402234) |
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| 摘 要： | 为拓宽噬菌体宿主谱,本研究通过将产肠毒素大肠杆菌(Enterotoxigenic Escherichia coli,ETEC)特异性噬菌体v B_Eco M_JS09与肠出血性大肠杆菌(Enterohemorrhagic Escherichia coli,EHEC)O157∶H7细菌进行多轮共培养,筛选到了6株能够同时裂解ETEC和EHEC O157∶H7的噬菌体,并多次传代和宿主谱鉴定,选择其中1株噬菌体Bp47-12鉴定其生物学特性。双层琼脂平板试验结果表明,噬菌体Bp47-12在平板上均呈现出了典型的裂解性噬菌体特征,空斑透亮,边缘整齐清晰,无晕环噬菌斑;宿主谱鉴定结果表明,ETEC和EHEC O157∶H7菌株均对噬菌体Bp47-12敏感,实现了噬菌体在ETEC和EHEC O157∶H7菌株之间的交叉裂解反应,拓宽了亲本噬菌体JS09的宿主谱;噬菌体Bp47-12能够耐受60℃左右高温,在p H值3~9时效价稳定;最佳感染复数为1;潜伏期为30 min,爆发期为70 min,平均爆发量分别为15;体外裂解试验结果表明,在5 h之内噬菌体Bp47-12能够有效裂解ETEC和EHEC O157∶H7菌株,OD600值下降约0.66;吸附曲线结果表明,在25 min之内,效价为105 PFU/mL的噬菌体Bp47-12能够完全吸附到宿主菌表面。本研究具有潜在的应用价值,也为拓宽裂解性噬菌体宿主谱和应用噬菌体提供了科学依据。
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| 关 键 词： | 抗生素耐药性 大肠杆菌 噬菌体 拓宽宿主谱 |
EXTENSION AND CHARACTERIZATION OF HOST RANGE OF E. COLI SPECIFIC BACTERIOPHAGES |
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| ZHOU Yan,XUE Jin-xin,YE Min,ZHANG Hui,BAO Hong-duo,WANG Ran.EXTENSION AND CHARACTERIZATION OF HOST RANGE OF E. COLI SPECIFIC BACTERIOPHAGES[J].Chinese Journal of Animal Infectious Diseases,2020(1):8-15. |
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| Authors: | ZHOU Yan XUE Jin-xin YE Min ZHANG Hui BAO Hong-duo WANG Ran |
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| Institution: | (Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base,Ministry of Science and Technology,Institute of Food Safety and Nutrition,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;School of Chemistry&Life Science,Nanjing University Jinling College,Nanjing 210014,China) |
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| Abstract: | The Enterotoxigenic Escherichia coli(ETEC)specific bacteriophage vBEcoMJS09 and Enterohemorrhagic Escherichia coli(EHEC)O157:H7 bacteria were co-cultured for several rounds in order to extend the host range of bacteriophages.Total six strains of bacteriophages were isolated and passaged several times for further selection.As a result,a strain of bacteriophage with broad host range was identified and designated as strain Bp47-12 for analysis of the biological characteristics.The results of double layer agar plate experiment revealed that Bp47-12 showed typical characteristics of lytic phage.The plaques were bright with neat and clear edges and no halo-rings around the plaques formed.The host range results showed that both ETEC and EHEC O157:H7 strains were sensitive to Bp47-12,which indicated the cross-lytic activity and extension of the host range of parental phage JS09.Bp47-12 tolerated high temperature of about 60℃,and was stable at pH 3-9.The results also showed the optimal MOI at 1,the latent period for 30 min,the burst period for 70 min,and the average burst size at 15 phages per cell.In vitro lytic assay revealed that Bp47-12 effectively lysed ETEC and EHEC O157:H7 strains in 5 h,and the OD600 value of bacteria decreased by about 0.66.The adsorption curve showed that Bp47-12 with potency of 10^5 PFU/mL completely adsorbed to the surface of host bacteria within 25 min. |
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| Keywords: | Antibiotic resistance E coli bacteriohage host range |
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