沙雷菌裂解性噬菌体PS2生物学特性及其全基因组测序

发布时间：2018-07-20 19:13

【摘要】：目的明确一株粘质沙雷菌裂解性噬菌体PS2的生物学特性及全基因组序列。方法电镜观察PS2的形态;双层平板法分析PS2噬菌谱、生长曲线及理化稳定性;采用MiniBEST Viral RNA/DNA Extraction Kit提取噬菌体基因组,酶解作用鉴定基因组类型;构建噬菌体PS2测序文库,采用MiSeqTM系统测序,Velvet 1.2.08进行序列拼接。结果 PS2为肌尾型噬菌体。其感染S2菌株的潜伏期为21min,爆发量为70PFU。PS2在pH5到pH10的环境中具有较好的稳定性,在50℃和60℃的环境中活性也无明显变化。PS2基因组为双链DNA,全长167 266bp,平均GC含量为41.7%,共预测到276个ORF。BLASTN初步比对结果提示其与多株T4-like噬菌体基因组核酸序列具有显著相似性。结论分离鉴定了一株粘质沙雷菌属噬菌体,进行了生物学特性、全基因组测序和生物信息学初步分析,为噬菌体治疗多重耐药细菌及噬菌体生物信息学深入分析奠定了基础。
[Abstract]:Objective to identify the biological characteristics and genome sequence of a lytic phage PS2 from Shareh. Methods the morphology of PS2 was observed by electron microscope, the phage spectrum, growth curve and physicochemical stability of PS2 were analyzed by double-layer plate method, the phage genome was extracted by MiniBEST virus RNA-DNA extraction kit, the genome type was identified by enzymatic hydrolysis, and the phage PS2 sequencing library was constructed. MiSeqTM system was used to sequence Velvet 1.2.08. Results PS2 was a muscular tail phage. The incubation period of S2 strain was 21 min, and the outbreak amount of 70PFU.PS2 was stable at pH 5 to pH10. There was no significant change in activity at 50 鈩，

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