RNA-seq analysis ofRv3574,Rv0047c,Rv0602c over-expression under 6h induction of ATc in Mycobacterium smegmatis

These vectors were then electroporated into Mycobacterium smegmatis and used as a control for pUV15 empty vector transfer. There were three experimental groups and one empty vector control group, with three biological replicates in each group and a total of 12 samples.

Cultivate the expression strain based on 37 ° C with 7H9 until the OD600 is around 0.4, then add ATc induced transcription factor at a final concentration of 50ng/mL and induce expression at 37 ° C for 6 hours. Subsequently, collect the bacteria under low temperature conditions, grind and crush them with liquid nitrogen, and extract RNA using triazol (Invitogen) according to the manufacturer's instructions. Subsequently, ribosomal RNA was removed using Ribo clean rRNA Deletion Kit Mega (Vazyme).