Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.
Laboratory of Phytopathology, Wageningen University, Wageningen, The Netherlands.
Sci Rep. 2017 Mar 7;7:43889. doi: 10.1038/srep43889.
We present integration vectors for Staphylococcus aureus encoding the fluorescent reporters mAmetrine, CFP, sGFP, YFP, mCherry and mKate. The expression is driven either from the sarA-P1 promoter or from any other promoter of choice. The reporter can be inserted markerless in the chromosome of a wide range of S. aureus strains. The integration site chosen does not disrupt any open reading frame, provides good expression, and has no detectable effect on the strains physiology. As an intermediate construct, we present a set of replicating plasmids containing the same fluorescent reporters. Also in these reporter plasmids the sarA-P1 promoter can be replaced by any other promoter of interest for expression studies. Cassettes from the replication plasmids can be readily swapped with the integration vector. With these constructs it becomes possible to monitor reporters of separate fluorescent wavelengths simultaneously.
我们提供了用于金黄色葡萄球菌的整合载体，该载体编码荧光报告蛋白 mAmetrine、CFP、sGFP、YFP、mCherry 和 mKate。表达由 sarA-P1 启动子或任何其他所需启动子驱动。报告基因可以无标记地插入到多种金黄色葡萄球菌菌株的染色体中。选择的整合位点不会破坏任何开放阅读框，提供良好的表达，并且对菌株的生理没有可检测的影响。作为中间构建体，我们提供了一组包含相同荧光报告蛋白的复制质粒。在这些报告质粒中，sarA-P1 启动子也可以被任何其他感兴趣的启动子替代，用于表达研究。复制质粒的盒可以很容易地与整合载体交换。有了这些构建体，就可以同时监测不同荧光波长的报告基因。
