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. 2011 Feb 9;39(11):4653–4663. doi: 10.1093/nar/gkr055

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© The Author(s) 2011. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Mapping AS promoters. (A) Overnight growth of E. coli cells harboring plasmids with AS promoters fused to promoterless galK gene on a McConkey agar plate. The following plasmids were used: pAS-galK—a DNA fragment (from +252 to +178) of Eco29kI system fused to promoterless galK; pASmut1-galK—pAS-galK with inactive AS_P1; pASmut2-galK—pAS-galK with inactive AS_P2; pASmut3-galK—pAS-galK with inactive AS_P1 and AS_P2. (B) The results of primer extension reaction carried out with RNA purified from cells harboring a plasmid with AS promoters fused to promoterless galK gene are shown. (C) Primer extension analysis of RNA samples purified from cells harboring a plasmid with intact Eco29kI system (p29), left lane, and the same plasmid with inactive eco29kIR promoters (p29-ResPmut, right lane) with primers designed to reveal AS promoters initiated transcripts.