Pseudomonas aeruginosa is an increasingly prevalent pathogen that has become a serious health concern due to an increasing incidence of multidrug-resistant (MDR) hospital-acquired infections. The emergence of MDR-P. aeruginosa coupled with shrinking antibiotic pipelines has increased the demand for new antimicrobials and therapeutics. An effective tool for drug screening both in vitro and in vivo can facilitate the discovery of drugs and regimens for treating P. aeruginosa infection. Here, for the first time, we combined the mini-Tn7 system and Xer/dif recombinase system to construct a stable and selectable marker-free autoluminescent P. aeruginosa (SfAlPa) by one step. Afterwards, in vitro and in vivo activities of several antibiotics including amikacin, biapenem, levofloxacin and polymyxin B were assessed using SfAlPa. This study demonstrated that the use of SfAlPa could significantly facilitate rapid real-time evaluating the activities of compounds. Compared to prevailing methods, this method reduces the time, effort, animals and costs consumed in the discovery of new drugs against P. aeruginosa. Additionally, the methodology described in this study could be easily modified for construction of selectable marker-free reporter strain in other Gram-negative bacteria.

铜绿假单胞菌（Pseudomonas aeruginosa）是一种日益普遍的病原体，由于医院获得的多药耐药性（MDR）感染的发生率增加，已成为严重的健康问题。MDR-铜绿假单胞菌的出现以及抗生素管道的减少增加了对新型抗菌剂和治疗剂的需求。一个有效的工具，药物筛选都在体外和体内可以促进药物和疗法的发现用于治疗铜绿假单胞菌感染。在这里，第一次，我们联合迷你Tn7位系统的Xer / DIF重组酶系统构建一个稳定和小号总统当选标记- ˚F REE一反对派升uminescent P。只需一步就可以制作出一种欧洲菊（SfAlPa）。之后，使用SfAlPa评估了几种抗生素（包括丁胺卡那霉素，比阿培南，左氧氟沙星和多粘菌素B）的体外和体内活性。这项研究表明，使用SfAlPa可以大大促进快速实时评估化合物的活性。与现有方法相比，该方法减少了发现铜绿假单胞菌新药的时间，精力，动物和成本。此外，本研究中描述的方法可以轻松修改，以构建其他革兰氏阴性细菌中的无标记选择性报告基因菌株。