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尾丝蛋白在噬菌体吸附鸭疫里默氏菌中的作用

刘娜 黄瑜 陈红梅 江南松 刘荣昌 傅光华 傅秋玲 梁齐章 万春和 李昂 程龙飞

刘娜,黄瑜,陈红梅,等. 尾丝蛋白在噬菌体吸附鸭疫里默氏菌中的作用 [J]. 福建农业学报,2024,39(1):1−6 doi: 10.19303/j.issn.1008-0384.2024.01.001
引用本文: 刘娜,黄瑜,陈红梅,等. 尾丝蛋白在噬菌体吸附鸭疫里默氏菌中的作用 [J]. 福建农业学报,2024,39(1):1−6 doi: 10.19303/j.issn.1008-0384.2024.01.001
LIU N, HUANG Y, CHEN H M, et al. Role of Tail Fiber Protein in Attaching Riemerella anatipestifer to Phage [J]. Fujian Journal of Agricultural Sciences,2024,39(1):1−6 doi: 10.19303/j.issn.1008-0384.2024.01.001
Citation: LIU N, HUANG Y, CHEN H M, et al. Role of Tail Fiber Protein in Attaching Riemerella anatipestifer to Phage [J]. Fujian Journal of Agricultural Sciences,2024,39(1):1−6 doi: 10.19303/j.issn.1008-0384.2024.01.001

尾丝蛋白在噬菌体吸附鸭疫里默氏菌中的作用

doi: 10.19303/j.issn.1008-0384.2024.01.001
基金项目: 福建省科技计划公益类专项(2021R10260011);福建省农业高质量发展超越“5511”协同创新工程项目(XTCXGC2021012);福建省农业科学院科技创新团队建设项目(CXTD2021005-1)
详细信息
    作者简介:

    刘娜(1997 —),女,硕士研究生,主要从事预防兽医学研究,E-mail:liunaupip560@163.com

    通讯作者:

    李昂(1964 —),男,硕士,教授,主要从事动物遗传育种与繁殖研究,E-mail:poul@fafu.edu.cn

    程龙飞(1971 —),男,硕士,研究员,主要从事家禽传染病学研究,E-mail:13705991428@139.com

  • 中图分类号: S855.1

Role of Tail Fiber Protein in Attaching Riemerella anatipestifer to Phage

  • 摘要:   目的  以鸭疫里默氏菌( Riemerella anatipestifer )噬菌体CRP2为对象,研究尾丝蛋白(Tail fiber protein,TFP)对噬菌体吸附功能的影响。  方法  克隆ORF70假定的TFP基因,构建重组表达载体pET28a(+)-TFP,在大肠杆菌E.Coli BL21(DE3)中诱导表达,通过Ni-NTA亲和层析纯化。酶联免疫吸附试验(ELISA)测定重组蛋白与CRP2噬菌体的宿主菌外膜蛋白(Outer membrane protein, OMP)的结合作用。竞争吸附试验测定重组蛋白对CRP2噬菌体吸附率的影响。  结果  构建了pET28a(+)-TFP重组表达质粒,在大肠杆菌中得到了可溶性表达。纯化的重组蛋白可以与宿主菌的OMP结合,也可以抑制CRP2对其宿主菌的吸附。  结论  ORF70编码的TFP是噬菌体CRP2的一种受体结合蛋白,其对应的受体是宿主菌的OMP。
  • 图  1  重组蛋白的SDS-PAGE分析

    M:蛋白Marker;1:TFP未诱导;2:TFP裂解液上清;3:TFP裂解液沉淀;4:纯化的rTFP。

    Figure  1.  SDS-PAGE of recombinant protein

    M: Protein molecular weight marker; 1: TFP expression without induction; 2: Expression of TFP lysate supernatant; 3: Expression of TFP lysate precipitate; 4: Purified protein of rTFP.

    图  2  重组蛋白的Western blotting分析

    1:rTFP;M:蛋白Marker。

    Figure  2.  Recombinant protein identified by western blot

    1: Recombinant rTFP; M: Protein molecular weight marker.

    图  3  重组蛋白与OMP的ELISA结果

    OMP+rTFP:OMP与rTFP的作用结果;OMP+PBS:OMP与PBS的作用结果。***:两组数据差异极显著(P<0.01)。

    Figure  3.  ELISA results on recombinant protein and OMP

    OMP+rTFP: Interaction between OMP and rTFP; OMP+PBS: Interaction between OMP and PBS. ***: extremely significant difference between two data sets( P <0.01).

    图  4  重组蛋白对噬菌体CRP2吸附率的影响

    Figure  4.  Effect of recombinant proteins on CRP2 adsorption rate

    图  5  噬菌体CRP2、CRP5、CRP12、CRP19尾丝蛋白氨基酸序列的比较

    图形上的黑色矩形框表示序列相似性最高,红色次之,绿色更低。黑点表示氨基酸缺失。

    Figure  5.  Amino acid sequences of TFPs in CRP2, CRP5, CRP12, and CRP19

    Black rectangle box indicates highest sequence similarity; red, intermediate; and green, lowest. Black dot indicates missing amino acid.

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出版历程
  • 收稿日期:  2023-09-17
  • 修回日期:  2023-11-14
  • 网络出版日期:  2024-01-25
  • 刊出日期:  2024-01-28

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