Depression of biofilm formation and antibiotic resistance by sarA disruption in Staphylococcus epidermidis

瞄准：到学习，简历上的 sarA 基因的混乱的效果拍摄形成和葡萄球菌 epidermidis 的抗菌素抵抗(S。epidermidis ) 。方法：以便破坏 sarA 基因，双转线路相应再结合在 S 被使用。epidermidis RP62A，和四圜素抵抗基因(tet ) 被用作被 PCR 从 pBR322 放大并且插入了到在在上游、下游的 sarA 之间的地点的选择标记，导致 pBT2delta sarA。由 electroporation，原生质标志 pBT2delta sarA 被转变成 S。epidermidis。基因抄写被即时反向的 transcription-PCR (RT-PCR ) 检测。简历电影的决心在 96 井被执行平底的培养皿，和抗菌素抵抗被分光光度测定法分别地在 570 nm 与试管文化分析。结果：破坏的 sarA 拉紧命名 S。epidermidis RP62Adelta sarA 被构造，它在简历电影形成是完全有缺点的，当 sarA 补充紧张 RP62Adelta sarA (pHPS9sarA ) 恢复了简历电影形成显型时。另外， sarA 的大美人导致了减少的抗生素的一种和 S 的卡那霉素抵抗。epidermidis RP62A。比作原来的紧张， S。epidermidis RP62Adelta sarA 分别地在 200-800 microg/mL 在 200-400 microg/mL 和卡那霉素有敏感的增加到抗生素的一种。结论：sarA 的大美人能在 S 在简历电影形成和减少的抗生素的一种和卡那霉素抵抗导致缺点。epidermidis RP62A。

AIM： To study the effects of disruption of sarA gene on biofilm formation and antibiotic resistance of Staphylococcus epidermidis （ S. epiderrnidis）. METHODS： In order to disrupt sarA gene, the double- crossover homologous recombination was applied in S. epiderrnidis RP62A, and tetracycline resistance gene （tet） was used as the selective marker which was amplified by PCR from the pBR322 and inserted into the locus between sarA upstream and downstream, resulting in pBT2ΔsarA. By electroporation, the plasmid pBT2ΔsarA was transformed into S. epiderrnidis. Gene transcription was detected by real-time reverse transcription-PCR （RT-PCR）. Determination of biofilm was performed in 96-well flat-bottomed culture plates, and antibiotic resistance was analyzed with test tube culture by spectrophotometry at 570 nm respectively. RESULTS： A sarA disrupted strain named S. epiderrnidis RP62AΔsarA was constructed, which was completely defective in biofilm formation, while the sarA complement strain RP62AΔsarA （pHPS9sarA） restored the biofilm formation phenotype. Additionally, the knockout of sarA resulted in decreased erythromycin and kanamycin resistance of S. epiderrnidis RP62A. Compared to the original strain, S. epiderrnidis RP62AΔsarA had an increase of the sensitivity to erythromycin at 200-400 μg/mL and kanamycin at 200-800 μg/mL respectively. CONCLUSION： The knockout of sarA can result in the defect in biofilm formation and the decreased erythromycin and kanamycin resistance in S. epiderrnidis RP62A.