Extended Data Fig. 4: Generation of BA metabolic pathway mutants in Bacteroides.
From: Microbial bile acid metabolites modulate gut RORγ+ regulatory T cell homeostasis
a, Schematic diagram of pNJR6 suicide vector-mediated BA gene deletion in Bacteroides. b, Genotyping of B. thetaiotaomicron and B. fragilis BA metabolic pathway mutants by PCR. PCR primers were designed to target the flanking regions of an intact gene. PCR of an untouched gene plus its flanking regions generated a PCR product of around 1,150–1,500 bp, while deletion of an interested BA metabolic gene resulted in only an approximately 350–450-bp PCR amplicon of its two flanking regions. c, d, Bacterial load (measured as colony-forming unit (CFU) per gram of faeces) of B. thetaiotaomicron (c) and B. fragilis (d) BA metabolic pathway mutants and their wild-type control strains in monocolonized GF mice. e, f, LC–MS quantification of faecal conjugated primary BAs (e) and deconjugated primary BAs (f) in GF mice monocolonized with B. thetaiotaomicron or B. fragilis BA metabolic pathway mutants and their wild-type control strains. Data are representative of two independent experiments in b, e and f. Data are pooled from three independent experiments in c and d. n represents biologically independent animals. Data are mean ± s.e.m. (c–f).
