霍乱弧菌分型噬菌体VP2部分生物学特性及基因组分析

孙惠惠 范宇峰 李臻鹏 赵硕 王晓勋 逄波 阚飙

孙惠惠, 范宇峰, 李臻鹏, 赵硕, 王晓勋, 逄波, 阚飙. 霍乱弧菌分型噬菌体VP2部分生物学特性及基因组分析[J]. 疾病监测. doi: 10.3784/jbjc.202105190282
引用本文: 孙惠惠, 范宇峰, 李臻鹏, 赵硕, 王晓勋, 逄波, 阚飙. 霍乱弧菌分型噬菌体VP2部分生物学特性及基因组分析[J]. 疾病监测. doi: 10.3784/jbjc.202105190282
Sun Huihui, Fan Yufeng, Li Zhenpeng, Zhao Shuo, Wang Xiaoxun, Pang Bo, Kan Biao. Partial biological characteristics and genomic analysis of Vibrio cholerae typing phage VP2[J]. Disease Surveillance. doi: 10.3784/jbjc.202105190282
Citation: Sun Huihui, Fan Yufeng, Li Zhenpeng, Zhao Shuo, Wang Xiaoxun, Pang Bo, Kan Biao. Partial biological characteristics and genomic analysis of Vibrio cholerae typing phage VP2[J]. Disease Surveillance. doi: 10.3784/jbjc.202105190282

霍乱弧菌分型噬菌体VP2部分生物学特性及基因组分析

doi: 10.3784/jbjc.202105190282
基金项目: 国家重点基础研究发展计划(No. 2015CB554201)
详细信息
    作者简介:

    孙惠惠,女,山东省诸城市人,博士研究生在读,主要从事霍乱弧菌分型噬菌体相关研究工作,Email:sunhuihui@nieh.chinacdc.cn

    通讯作者:

    阚飙,Tel:010–58900742,Email:kanbiao@icdc.cn

  • 中图分类号: R211; R378.3

Partial biological characteristics and genomic analysis of Vibrio cholerae typing phage VP2

Funds: This study was supported by the National Basic Science and Technology Fund (No. 2015CB554201)
More Information
  • 摘要:   目的  对霍乱弧菌分型噬菌体VP2基因组、蛋白质组和增殖特征等开展分析,获取VP2的生物学特征。  方法  电镜观察噬菌体VP2的形态,测定最佳感染复数和一步生长曲线;通过在线RAST(https://rast.nmpdr.org/)对噬菌体VP2基因组进行编码基因预测及功能注释;提取VP2成熟颗粒中的蛋白质进行质谱分析;使用pyani(https://github.com/widdowquinn/pyani)进行噬菌体全基因组平均核苷酸一致性(ANI)聚类分析。  结果  噬菌体VP2为典型的20面体短尾噬菌体,最佳感染复数为1∶100,一步生长曲线结果显示其潜伏期约为60 min,60~120 min为爆发期,120 min后为稳定期。 预测存在49个开放阅读框,VP2成熟颗粒蛋白质谱分析显示有34个蛋白与预测的基因相对应,ANI聚类分析结果显示VP2与弧菌噬菌体CJY的ANI最高。  结论  明确了分型噬菌体VP2的形态和基因组特征,预测的功能蛋白及结构蛋白的鉴定为后续研究同源噬菌体及其与霍乱弧菌的相互作用奠定了基础。
  • 图  1  VP2噬菌体在电镜下形态(×97 000)

    注:红色箭头示短尾

    Figure  1.  Morphology of VP2 phage under transmission electron microscopy (the short tail of the phage is shown at the red arrow) (×97 000)

    图  2  噬菌体VP2的一步生长曲线

    Figure  2.  One-step growth curve of phage VP2

    图  3  VP2全基因组模式图

    注:最外层表示噬菌体基因组全长,第二圈代表正向阅读框;第三圈代表负向阅读框,其中蓝色为功能基因,橙色为结构基因,绿色为假定基因;第四圈黄色与绿色代表GC含量,其中黄色表示低于全基因组平均GC含量,绿色则相反;最内圈玫红色表示GC偏嗜性G-C/G+C<0,蓝色表示>0

    Figure  3.  Genome map of phage VP2

    图  4  VP2全基因组核苷酸一致性分析

    Figure  4.  Average nucleotide identity of VP2

    表  1  VP2噬菌体与宿主菌16017的最佳感染复数

    Table  1.   The optimal MOI between VP2 phage and bacteria 16017

    噬菌体滴度宿主菌浓度MOI(噬菌体∶菌)结果
    1061061∶1 2.00×106
    1051061∶10 5.20×107
    1041061∶1006.80×107
    下载: 导出CSV

    表  2  VP2颗粒全蛋白质谱与VP2全基因组测序预测蛋白14比对结果

    Table  2.   Comparison between the results of VP2 granule protein spectrum and the predicted proteins of VP2 whole genome sequencing14

    VP2ORF起始基因位点终止基因位点氨基酸长度(aa)功  能蛋白覆盖度(%)
    1409630222  hypothetical protein
    26471147501  terminase small subunit36
    3113428401707  terminase15
    4285144941644  Head-tail connecting protein78
    544944721228  hypothetical protein96
    647344991258  hypothetical protein
    749965838843  hypothetical protein58
    860066974969  structural protein84
    970417310270  hypothetical protein97
    1073258173849  hypothetical protein77
    1181708535366  hypothetical protein59
    1285379010474  LysM peptidoglycan-binding domain-containing protein27
    1390039305303  Phage protein93
    149329115542226  PE family protein/tail protein76
    1511563133471785  structural protein76
    161335313757405  Phage protein98
    1713754158382085  Phage protein93
    1815838170101173  hypothetical protein78
    1917012193602349  structural protein79
    2019364204371074  Phage tail fibers54
    212043921821366  Outer capsid protein53
    222222021846474  hypothetical protein
    232267722240303  hypothetical protein45
    2422828226672226  hypothetical protein
    2523022228251785  metal dependent phosphohydrolase65
    262333623019405  Hydrolase91
    2723598233382085  hypothetical protein
    2824631236001032  Adenylosuccinate synthetase71
    2926961246732289  integrase54
    3028819269511869  DNA polymerase I17
    312933728861477  Single stranded DNA-binding protein,phage-associated15
    323021229403810  hypothetical protein29
    333080030333468  hypothetical protein17
    3432332308631470  superfamily II DNA/RNA helicase29
    353275032397354  hypothetical protein17
    363344232750693  hypothetical protein20
    373383233575258  hypothetical protein18
    383444634036411  structural protein
    393477434448327  hypothetical protein
    403562434998627  hypothetical protein14
    413582835667162  hypothetical protein
    423641835933486  hypothetical protein
    433685936428432  Phage protein38
    443740737036372  hypothetical protein
    453763137407225  hypothetical protein
    463799937709291  hypothetical protein
    473834538106240  hypothetical protein
    483894638356591  hypothetical protein
    493981739548270  hypothetical protein
    下载: 导出CSV
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  • 收稿日期:  2021-05-19
  • 网络出版日期:  2021-09-16

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