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. 2007 Mar 13;35(6):2060–2073. doi: 10.1093/nar/gkm005

Figure 3.

Figure 3.

Radioactive reverse transcription PCR (RT-PCR) analysis of strain SSB318 complemented with S. aureus rnpBwt or rnpBC238/C239. PCR products were analyzed on a 10% polyacrylamide/8 M urea gel. Lanes 1–30: total RNA from SSB318 complemented with S. aureus rnpBwt (lanes 1–4 and 13-16), rnpBC238 (lanes 5–8, 17–20 and 25–30) or rnpBC239 (lanes 9–12 and 21–24) grown at 37°C in the absence of IPTG and in the presence of 2% xylose (w/v); amounts of total RNA were 200 ng in lanes 1–24, 26 and 29, 100 ng in lanes 25 and 28, and 400 ng in lanes 27 and 30. Pxyl rnpA: presence (+) or absence (−) of a xylose-inducible plasmid-encoded B. subtilis rnpA gene. Lanes 1–12 and 25–27: primers specific for S. aureus rnpB (rnpB); lanes 13–24 and 28–30: primers specific for the mRNA encoding B. subtilis ribosomal protein S18 (S18). AMV: presence (+) or absence (−) of reverse transcriptase. For details on RT-PCR, see the Material and Methods section. Lanes 25–30 document that the amount of RT-PCR product was sensitive to RNA template concentration. The figure illustrates a representative experiment, but the results shown here were reproduced in five individual experiments using three independent total RNA preparations.