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. 2001 Apr;183(8):2476–2484. doi: 10.1128/JB.183.8.2476-2484.2001

FIG. 1.

FIG. 1

Generation of miniplasmids from pSEK131. (A) Ethidium bromide-stained agarose gel (0.7%) showing miniplasmids in various E. coli strains harboring pSEK131. Lane 1, YK1100 (wild type [wt]); lane 2, YK1340 (hupA hupB); lane 3, YK2920 (himA hip); lane 4, YK2741 (himA hupA hupB); lane 5, YK4122 (wt); lane 6, YK4124 (hns); lane 7, YK4137 (hns hupA hupB); lane 8, YK4205 (hns himA hip); lane 9, YK5246 (stpA); lane 10, YK5244 (stpA hns); lane 11, RZ4500 (wt); lane 12, MDW246 (fis); lane 13, CSH26 (wt); lane 14, CU211 (hns); lane 15, KT1008 (wt); lane 16, KT1004 (rpoS); lane 17, KT1008 hns (hns); lane 18, KT1004 hns (rpoS hns). Positions of pSEK131 (monomers [M] and dimers [D]) and several kinds of miniplasmids are indicated. ccc and oc indicate covalently closed circular and open circular DNA molecules, respectively. Note that miniplasmids of pSEK131 were generated in the hns wild-type strains (+) but not in the hns mutant strains (−). (B) Ethidium bromide-stained agarose gels (1%) showing PCR-amplified fragments from miniplasmids of pSEK131. PCR was performed with (a) the total plasmid DNA prepared from the same E. coli strains as those used above (lanes 1 to 18); (b) the total plasmid DNA prepared from strains CU241 (wt; lane 1) and CU242 (Δhns::Kmr; lane 2) and from the CU241 derivatives with an hns mutation in the N- or C-terminal domain, HM12 (hns12; lane 3), HM52 (hns52; lane 4), and HM60 (hns60; lane 5); and (c) the total plasmid DNA prepared from the H-NS-deficient strain YK4124 harboring plasmid pYS6 with the wild-type hns gene (lane 2), plasmid pYS7 with the hns gene mutated in the central domain (lane 3), or pSTV28 without the hns gene (lane 1). The DNA fragments marked with α indicate PCR fragments from pSEK131, and small fragments marked with β indicate those from the most abundant miniplasmids of pSEK131. Molecular sizes of linear marker DNA fragments are shown on the side of the gel. Note that PCR fragments from miniplasmids were generated in the hns mutant strain (−∗) with a mutation in the N- or C-terminal domain but not in the one with a mutation in the central domain.