Discovery, purification, and characterization of a temperate transducing bacteriophage for Bordetella avium.
Discovery, purification, and characterization of a temperate transducing bacteriophage for Bordetella avium.
复制标题
鸟博德特氏菌温带转导噬菌体的发现、纯化和表征。
DOI：
10.1128/jb.182.21.6130-6136.2000
复制
发表时间：
2000
影响因子：
3.2
通讯作者：
Orndorff,PE
中科院分区：
文献类型：
--
作者：
Shelton,CB;Crosslin,DR;Casey,JL;Ng,S;Temple,LM;Orndorff,PE
我们发现了一种温和的转导噬菌体（Ba 1）的禽呼吸道病原体禽波氏杆菌。在筛选了4株B.溶原性的疫苗。在这两种噬菌体中，只有Ba 1显示出通过等位基因置换机制进行噬菌体的能力，并对其进行了进一步的研究。就宿主范围而言，Ba 1在9株临床分离的B.但在支气管败血波氏杆菌、欣兹波氏杆菌、百日咳波氏杆菌或副百日咳波氏杆菌的任何测试菌株上均不能生长。Ba 1纯化CsCl梯度离心，并发现有一个二十面体的头部，其中包含一个线性基因组的约46.5 kb（轮廓长度）的双链DNA和收缩，鞘尾。Ba 1很容易溶原化我们的实验室。禽流感病毒株（197 N），在无外界感染的情况下，原噬菌体状态至少能稳定25代。DNA杂交研究表明，原噬菌体在宿主和噬菌体复制子上的优选位点整合。Ba 1转导五个明显不同的插入突变，表明转导是普遍的。转导频率范围约为2 × 10 - 7至1 × 10 - 8 transducants/PFU，取决于被转导的标记。UV照射的转导裂解物显着提高转导频率，并减少了转导过程中溶原化的转导子的数量。Ba 1可能被证明是研究B的有用的遗传工具。禽毒力因子
We discovered and characterized a temperate transducing bacteriophage (Ba1) for the avian respiratory pathogenBordetella avium. Ba1 was initially identified along with one other phage (Ba2) following screening of four strains ofB. aviumfor lysogeny. Of the two phage, only Ba1 showed the ability to transduce via an allelic replacement mechanism and was studied further. With regard to host range, Ba1 grew on six of nine clinical isolates ofB. aviumbut failed to grow on any tested strains ofBordetella bronchiseptica,Bordetella hinzii,Bordetella pertussis, orBordetella parapertussis. Ba1 was purified by CsCl gradient centrifugation and was found to have an icosahedral head that contained a linear genome of approximately 46.5 kb (contour length) of double-stranded DNA and a contractile, sheathed tail. Ba1 readily lysogenized our laboratoryB. aviumstrain (197N), and the prophage state was stable for at least 25 generations in the absence of external infection. DNA hybridization studies indicated the prophage was integrated at a preferred site on both the host and phage replicons. Ba1 transduced five distinctly different insertion mutations, suggesting that transduction was generalized. Transduction frequencies ranged from approximately 2 × 10−7to 1 × 10−8transductants/PFU depending upon the marker being transduced. UV irradiation of transducing lysates markedly improved transduction frequency and reduced the number of transductants that were lysogenized during the transduction process. Ba1 may prove to be a useful genetic tool for studyingB. aviumvirulence factors.