Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus
Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus
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DOI：
10.3791/55087
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发表时间：
2017-03-01
影响因子：
1.2
通讯作者：
Morikawa, Kazuya
中科院分区：
文献类型：
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作者：
Cafini, Fabio;Nguyen Thi Le Thuy;Morikawa, Kazuya
主要的人类机会致病菌金黄色葡萄球菌的一个重要特征是其快速获得抗生素抗性的非凡能力。基因组研究表明，S.金黄色葡萄球菌携带许多位于移动的遗传元件上的毒力和抗性基因，表明水平基因转移（HGT）在金黄色葡萄球菌中起关键作用。金黄色葡萄球菌进化然而，一个完整和详细的描述的方法来研究HGT在S。金黄色葡萄球菌仍然缺乏，特别是关于自然转化，这是最近在这种细菌中报道的。本工作描述了三个协议，是有用的HGT在体外调查S。金黄色葡萄球菌：接合、噬菌体转导和自然转化。为此，使用了cfr基因（氯霉素/氟苯尼考抗性），其赋予酚类、林可酰胺类、恶唑烷酮类、截短侧耳素类和链阳性菌素A（PhLOPSA）抗性表型。了解S.金黄色葡萄球菌将遗传物质转移到其他菌株对于理解快速获得耐药性至关重要，并有助于澄清监测计划中报告的传播模式或进一步预测未来的传播模式。
One important feature of the major opportunistic human pathogen Staphylococcus aureus is its extraordinary ability to rapidly acquire resistance to antibiotics. Genomic studies reveal that S. aureus carries many virulence and resistance genes located in mobile genetic elements, suggesting that horizontal gene transfer (HGT) plays a critical role in S. aureus evolution. However, a full and detailed description of the methodology used to study HGT in S. aureus is still lacking, especially regarding natural transformation, which has been recently reported in this bacterium. This work describes three protocols that are useful for the in vitro investigation of HGT in S. aureus: conjugation, phage transduction, and natural transformation. To this aim, the cfr gene (chloramphenicol/florfenicol resistance), which confers the Phenicols, Lincosamides, Oxazolidinones, Pleuromutilins, and Streptogramin A (PhLOPSA)-resistance phenotype, was used. Understanding the mechanisms through which S. aureus transfers genetic materials to other strains is essential to comprehending the rapid acquisition of resistance and helps to clarify the modes of dissemination reported in surveillance programs or to further predict the spreading mode in the future.