RNA-seq of  Clostridioides difficile  ΔsigV、Δanti-sigV against wild-type strain control
The WT, ΔsigV, and Δanti-sigV strains were inoculated into BHIS medium and cultured until an OD600 of 0.6 was reached. The cultures were then centrifuged at 13,500 × g for 10 min, the supernatant was discarded, and the bacterial pellets of each strain were collected. The samples were transported with sufficient dry ice in a foam box to Shanghai Majorbio Bio-Pharm Technology Co., Ltd. for transcriptome sequencing. Total RNA was extracted from WT, ΔsigV, and Δanti-sigV using a bacterial total RNA extraction kit (DP430, TIANGEN, Beijing). Once the samples were deemed satisfactory, the following steps were conducted for sequencing: (1) Removal of ribosomal RNA; (2) Enrichment and purification of mRNA; (3) Fragmentation of mRNA; (4) Construction of the sequencing library and quality assessment of the library.