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. 2008 Oct 1;36(19):6249–6259. doi: 10.1093/nar/gkn633

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© 2008 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 8. — Analysis of tisAB mediated uxaA cleavage. (A) Map of uxaCA operon and position of northern probes and antisense region. Drawing is to scale. (B) Expression and cleavage of uxaA mRNA in BK4012 (tisAB::Kan^r), AB1157 and AB1157 transformed with pBK410 (overexpressing tisAB). Total RNA was extracted from the three isogenic strains and 5 µg RNA obtained from unexposed cells (–) and cells exposed to UV (50 J/m²) at two different time points (5 and 20 min after UV exposure) was separated on formaldehyde-1% agarose gel and blotted onto a nylon membrane. The blot was hybridized with a 400 nt uxaA riboprobe (Probe 1) covering the uxaA/tisAB antisense sequence (lower panel), and rehybridized with a 1100 nt uxaA riboprobe (Probe 2) covering only the sequence upstream of the antisense sequence. Size marker: 4 µg Millennium Markers-Formamide (Ambion). (C) Sequence alignments indicating base changes of the pBK410 derived mutants tisABuA1 and tisABuA2 followed by cleavage of uxaA mRNA in AB1157 transformed with pBK410 and the antisense mutants tisABuA1 and tisABuA2. Northern was carried out as in (B) except the blot was hybridized only with the 400-nt probe (Probe 1).