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分枝杆菌噬菌体Leo的生物学和基因组学特征

发布时间:2018-05-13 16:58

  本文选题:分枝杆菌噬菌体Leo + 基因组 ; 参考:《华中科技大学学报(医学版)》2014年05期


【摘要】:目的筛选能有效裂解结核分枝杆菌的噬菌体,并了解其生物学和遗传学信息。方法双层平板法扩增噬菌体,观察噬菌斑形态;电镜观察噬菌体颗粒形态;单斑法检测噬菌体宿主谱。采用λ噬菌体提取试剂盒提取噬菌体DNA,鸟枪法测序,得到的DNA序列用Phred/PhraD/Consedh软件包组装,拼接重叠群,PCR扩增重叠群缺口。采用DNAStar软件包、Glimmer3.0基因预测软件、Promoter prediction/Prokaryotic程序进行基因成分分析,基因功能预测,启动子区预测,并同时对Leo作共线性分析及构建系统进化树。结果噬菌体Leo由多面体头部和尾部构成,头部直径(70.0±3.0)nm,尾长(211.0±31.7)nm。其噬菌斑清晰透明,能裂解不同的结核分枝杆菌临床分离株。Leo基因组全长39 981bp,GC含量66.86%,含3个可能的启动子区,不含串联重复序列和tRNA区。Leo含59个基因,gene32编码整合酶,不含编码阻遏蛋白的基因和不利基因,Leo与BPs和Angel相似性最高。结论噬菌体Leo是一株不含不利基因的裂解性噬菌体,能裂解结核分枝杆菌,可以作为鸡尾酒疗法的候选噬菌体。
[Abstract]:Objective to screen bacteriophages capable of effectively lysing Mycobacterium tuberculosis and to understand their biological and genetic information. Methods the phage was amplified by double plate method to observe the morphology of phage plaque, the morphology of phage particle was observed by electron microscope, and the host spectrum of phage was detected by single spot method. The bacteriophage DNA was extracted by 位 phage extraction kit and sequenced by bird gun method. The resulting DNA sequence was assembled by Phred/PhraD/Consedh software package, and overlapping group gap was amplified by splicing overlapping colony. Gene composition analysis, gene function prediction and promoter region prediction were carried out by DNAStar software package Glimmer 3.0 gene prediction software named Promoter prediction/Prokaryotic. At the same time, co-linear analysis of Leo and construction of phylogenetic tree were carried out. Results the bacteriophage Leo was composed of polyhedron head and tail, the head diameter was 70.0 卤3.0 nmand the tail length was 211.0 卤31.7nm. Its plaque is clear and transparent, and it can break down the full length of 39 981bpGC of different Mycobacterium tuberculosis isolates. Leo genome contains three possible promoter regions, no tandem repeat sequence and tRNA region containing 59 gene gene32 coding integrase. There was the highest similarity between Leo and BPs and Angel without encoding repressor protein. Conclusion Phage Leo is a lytic bacteriophage without unfavorable gene and can be used as a candidate phage for cocktail therapy.
【作者单位】: 重庆医科大学附属第一医院呼吸与危重症医学科;成都市第三人民医院呼吸内科;重庆市长寿区人民医院呼吸内科;
【基金】:国家“十二五”重大专项课题资助项目(No.2012ZX10003009) 国家临床重点专科专项经费资助项目(No.卫办医政函【2012】649号)
【分类号】:R378

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