CBASS_Experiment (CRX1830227; CRA027583)
M. smegmatis mc2155 strains containing the pYC601-cmaABC plasmid were inoculated into fresh 7H9 medium supplemented with hygromycin and 10 ng/mL ATc, with an initial OD600 of 0.03. The cultures were grown at 37 oC with shaking at 200 rpm until an OD600 of approximately 0.8. The competent cells were prepared and transformed with TM4 gene expression plasmids 36. After a recovery period of 4 hours, the cultures were plated onto 7H10 agar supplemented with hygromycin, zeocin, and 50 ng/mL ATc and incubated at 37 oC. A control group was prepared under the same conditions without the addition of ATc to induce defense system expression. The transformants were scraped from the plates, and plasmids were extracted for Next Generation Sequencing (NGS) and comparative analysis.
Institute of pathogen Biology Chinese Academy of Medical Sciences Peking Union Medical College
DRX796064
NGDC
CRX1830227