本文選題：分枝桿菌噬菌體Leo + 基因組　； 參考：《華中科技大學學報(醫(yī)學版)》2014年05期

【摘要】：目的篩選能有效裂解結核分枝桿菌的噬菌體,并了解其生物學和遺傳學信息。方法雙層平板法擴增噬菌體,觀察噬菌斑形態(tài);電鏡觀察噬菌體顆粒形態(tài);單斑法檢測噬菌體宿主譜。采用λ噬菌體提取試劑盒提取噬菌體DNA,鳥槍法測序,得到的DNA序列用Phred/PhraD/Consedh軟件包組裝,拼接重疊群,PCR擴增重疊群缺口。采用DNAStar軟件包、Glimmer3.0基因預測軟件、Promoter prediction/Prokaryotic程序進行基因成分分析,基因功能預測,啟動子區(qū)預測,并同時對Leo作共線性分析及構建系統(tǒng)進化樹。結果噬菌體Leo由多面體頭部和尾部構成,頭部直徑(70.0±3.0)nm,尾長(211.0±31.7)nm。其噬菌斑清晰透明,能裂解不同的結核分枝桿菌臨床分離株。Leo基因組全長39 981bp,GC含量66.86%,含3個可能的啟動子區(qū),不含串聯(lián)重復序列和tRNA區(qū)。Leo含59個基因,gene32編碼整合酶,不含編碼阻遏蛋白的基因和不利基因,Leo與BPs和Angel相似性最高。結論噬菌體Leo是一株不含不利基因的裂解性噬菌體,能裂解結核分枝桿菌,可以作為雞尾酒療法的候選噬菌體。
[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 鹵3.0 nmand the tail length was 211.0 鹵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.
【作者單位】： 重慶醫(yī)科大學附屬第一醫(yī)院呼吸與危重癥醫(yī)學科;成都市第三人民醫(yī)院呼吸內(nèi)科;重慶市長壽區(qū)人民醫(yī)院呼吸內(nèi)科;
【基金】：國家“十二五”重大專項課題資助項目(No.2012ZX10003009) 國家臨床重點�？茖ｍ椊�(jīng)費資助項目(No.衛(wèi)辦醫(yī)政函【2012】649號)
【分類號】：R378

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