Extended Data Fig. 6. Generation of gene deletion mutants of P. clara JCM14859, growth of mutants deficient in trypsin degradation and analysis of genes located between 00502 and 00509.
a, Schematic illustration (upper panel) and PCR validation (lower panels) of 03048-03053, 00502 and 00509 gene deletion. Primers used for mutagenesis and PCR validation are listed in Supplementary Table 5. pLGB30: suicide vector used for cloning and integrating sequences into P. clara JCM14859. b, Wild type (WT), Δ00502 or Δ00509 P. clara JCM14589 strains were incubated with recombinant mouse PRSS2 (rmPRSS2) and degradation of rmPRSS2 was analysed by Western Blot with anti-His-tag antibody. c, In vitro growth rate of mutants deficient in trypsin degradation as determined by OD600. : d, Alignment of the 00502-00509 gene cluster in Paraprevotella genomes and annotation of each protein with Prokka 1.14.6. e, Wild type (WT) or the indicated mutant strains of P. clara JCM14589 were incubated with rmPRSS2, and degradation of rmPRSS2 was analysed by Western Blot. c, Data shown as mean ± s.d. n = 5 wells of individual bacterial cultures per group. a, b, e, Images from one experiment are shown. See Supplementary Figure 1 for gel and blot source data.