Transfer of Plasmid DNA to Clinical Coagulase-Negative Staphylococcal Pathogens by Using a Unique Bacteriophage
Transfer of Plasmid DNA to Clinical Coagulase-Negative Staphylococcal Pathogens by Using a Unique Bacteriophage
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使用独特的噬菌体将质粒 DNA 转移至临床凝固酶阴性葡萄球菌病原体
DOI：
10.1128/aem.04190-14
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发表时间：
2015
影响因子：
4.4
通讯作者：
Rohde H
中科院分区：
文献类型：
--
作者：
Winstel V;Kühner P;Krismer B;Peschel A;Rohde H
新出现的细菌病原体，如凝固酶阴性葡萄球菌（CoNS）的基因操作是临床和基础微生物学研究的主要障碍。强大的遗传障碍，如限制性修饰系统或成簇的规则间隔短回文重复序列（CRISPR），通常会干扰DNA转化的可用技术，因此使CoNS的操作复杂化或使其不可能。因此，目前对CoNS的致病性和毒力决定因素的了解非常有限。在这里，提出了一种快速，高效，高度可靠的技术，将基因工程所必需的质粒DNA从独特的金黄色葡萄球菌菌株通过特定的S.金黄色葡萄球菌噬菌体，Φ187。一旦供体和受体菌株具有相似的Φ187受体特性，即使是电穿孔难治的菌株也可以通过该技术转导。作为原理证明，该技术用于通过等位基因置换以高效率删除鼻和临床表皮葡萄球菌分离株中的替代转录因子σ B（SigB）。所描述的方法将允许对广泛的CoNS病原体进行遗传操纵，并可能激发研究活动以类似的方式操纵其他重要病原体。
Genetic manipulation of emerging bacterial pathogens, such as coagulase-negative staphylococci (CoNS), is a major hurdle in clinical and basic microbiological research. Strong genetic barriers, such as restriction modification systems or clustered regularly interspaced short palindromic repeats (CRISPR), usually interfere with available techniques for DNA transformation and therefore complicate manipulation of CoNS or render it impossible. Thus, current knowledge of pathogenicity and virulence determinants of CoNS is very limited. Here, a rapid, efficient, and highly reliable technique is presented to transfer plasmid DNA essential for genetic engineering to important CoNS pathogens from a unique Staphylococcus aureus strain via a specific S. aureus bacteriophage, Φ187. Even strains refractory to electroporation can be transduced by this technique once donor and recipient strains share similar Φ187 receptor properties. As a proof of principle, this technique was used to delete the alternative transcription factor sigma B (SigB) via allelic replacement in nasal and clinical Staphylococcus epidermidis isolates at high efficiencies. The described approach will allow the genetic manipulation of a wide range of CoNS pathogens and might inspire research activities to manipulate other important pathogens in a similar fashion.
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影响因子：
16.6
作者：
Winstel, Volker;Liang, Chunguang;Sanchez-Carballo, Patricia;Steglich, Matthias;Munar, Marta;Broeker, Barbara M.;Penades, Jose R.;Nuebel, Ulrich;Holst, Otto;Dandekar, Thomas;Peschel, Andreas;Xia, Guoqing
通讯作者：
Xia, Guoqing
影响因子：
6.7
作者：
Geiger T;Francois P;Liebeke M;Fraunholz M;Goerke C;Krismer B;Schrenzel J;Lalk M;Wolz C
通讯作者：
Wolz C
影响因子：
2.6
作者：
Bae, T;Schneewind, O
通讯作者：
Schneewind, O
葡萄球菌管病毒和肌病毒的壁磷壁酸依赖性吸附
影响因子：
3.2
作者：
Guoqing Xia;R. Corrigan;Volker Winstel;C. Goerke;A. Gründling;A. Peschel
通讯作者：
A. Peschel
影响因子：
2.8
作者：
Heilbronner, Simon;Hanses, Frank;Foster, Timothy J.
通讯作者：
Foster, Timothy J.