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. 2011 Aug;79(8):3229–3238. doi: 10.1128/IAI.00019-11

Fig. 5.

Fig. 5.

Confirmation and gene expression of a C. difficile prsW insertion mutant. (A and B) Schematic representations of the wild-type prsW gene (A) and the prsW gene disrupted by the LtrAL1::ermB intron of C. difficile strain JIR8094 (B). The locations and orientations of the oligomers used for PCR screening (CDEP515, CDEP516, and CDEP692) are represented as arrows. (C) PCR analysis of the JIR8094 parent strain (wild type [wt]) and the prsW::ltrBL1::ermB (prsW mutant) strain. The oligomers CDEP515 and CDEP516 generated a 465-bp product for the wild type and a 2,400-bp product for the prsW mutant. The oligomers CDEP515 and CDEP692 amplified no band for the wild type and a 435-bp product for the prsW mutant. The outside lanes contain a 1-kb DNA ladder from NEB. (D) Relative RNA levels of the different genes in the wild type or the prsW mutant. The oligonucleotides used in the qRT-PCR for csfT (TEQ003 and TEQ004), csfU (TEQ005 and TEQ006), csfV (TEQ007 and TEQ008), tcdA (TEQ017 and TEQ018), and rpoB (TEQ009 and TEQ010) are listed in Table S1 in the supplemental material. RNA levels were normalized to the housekeeping gene rpoB. Experiments were performed in triplicate, and the standard deviations are indicated.