• BMB Reports
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• v.29 no.1
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• pp.63-67
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• 1996

In order to elucidate the molecular mechanism of plasmid incompatibility of broad host-range plasmid R1162, the plasmid-encoded replication protein RepIB was purified and tested for binding to the 20 bp direct repeat (DR) DNA sequence which is reiterated 3 and 1/2 times within the replicative origin of the plasmid. The RepIB protein specifically binds to the DR DNA. Point mutations in the DR which affect expression of plasmid incompatibility also coordinately affect binding. These results indicate that the incompatibility of broad host-range plasmid R1162 is exerted by the DR DNA by titrating the essential replication protein RepIB.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1614-1620
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• 2015

Plasmid-cured derivative strains of Bacillus anthracis are frequently used in laboratory studies. Plasmid incompatibility, which does not increase the risk of chromosomal mutation, is a useful method for plasmid curing. However, in bacteria containing multiple plasmids, it often requires the sequential introduction of multiple, specific incompatibility plasmids. This lengthy process renders the traditional plasmid incompatibility method inefficient and mutation-prone. In this study, we successfully cured plasmids pXO1 and pXO2 from B. anthracis A16 simultaneously using only one recombinant incompatible plasmid, pKORT, to obtain a plasmid-free strain, designated A16DD. This method may also be useful for the simultaneous, one-step curing of multiple plasmids from other bacteria, including Bacillus thuringiensis and Yersinia pestis.

• Microbiology and Biotechnology Letters
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• v.17 no.1
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• pp.51-55
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• 1989

The conjugally transferred TOL plasmid or NAH plasmid was stably maintained and expressed in n-alkane assimilating Pseudomonas putida KCTC 2405. However, these plasmids were not able to coexist in this strain because of incompatibility. The incompatibility of TOL and NAH plasmid was bypassed using CAM::TOL* plasmid, which was constructed by the transposition of only tol gene without incompatibility system in TOL plasmid into CAM plasmid. p. putida 3SK capable of growing on m-toluate, naphthalene, camphor, and n-alkane(C8-C24) was constructed by the conjugal transfer of NAH plasmid into n-alkane assimilating p. putida SK carrying CAM:: TOL* plasmid. CAM::TOL＊ plasmid in p. putida 3SK was stable on the selective media but unstable on the nonselective media.

• Korean Journal of Microbiology
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• v.27 no.4
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• pp.334-341
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• 1989

Toluate degradative plasmid from Pseudomonas cepacia SUB37 was determined to be molecular weight as $79.3\times 10^6$

• Korean Journal of Microbiology
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• v.25 no.2
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• pp.87-93
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• 1987

The properties of the plasmid pKU41 isolated from Pseudomonas putkda KU190 have been investigated, pKU41 was defined as an R plasmid having a transmissible ampicillin and tetracycline resistance determinant, and could be classified as a plasmid belonging to IncP-1 group according to incompatibility grouping.

• Korean Journal of Veterinary Research
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• v.29 no.2
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• pp.59-67
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• 1989

This paper deals with the genetic properties of R plasmids in Salmonella originated from pigs and cattle. The plasmid DNA was examined for incompatibility, stability and fertility inhibition(Fi), and gel electrophoresis was performed for isolation of plasmid DNA. The results obtained were summerized as follows: 1. Among the 66 conjugative R plasmids from 44 pigs and 22 cattle, 61 R plasmids (92.4%) were $Fi^-$, whereas the remainder were $Fi^+$. 2. The Inc groups of 66 R plasmids were determined with 7 standard plasmids. Twenty-six R plasmids were classified into Inc group $I{\alpha}$, H1, H2 or F1, 40 R plasmids being not classified with standard plasmids used, and the Inc group $I{\alpha}$ (57.7%) was most frequent. 3. Inc groups $I{\alpha}$, H1, and F1 were identified in strains from swine, Inc groups H2 and F1 from cattle. 4. The plasmid DNA profiles in 16 Salmonella isolated from pigs and cattle were confirmed as being 1 to 10 fragments by the gel eletrophoresis. Their molecular weight ranged 1.0 to 90 megadalton. 5. The molecular weight of conjugative plasmids ranged 1.0 to 80 megadalton in 4 Salmonella (P-4, P-5, P-7 and P-8) isolated from pigs.

• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.433-436
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• 1990

The TOL plasmid, pWWO, conjugally transferred from Pseudomonas putida mt-2 was dissociated into TOL* and TOL $\Delta$A in P. putidu PpGl carrying CAM plasmid. The TOL* was integrated into the CAM plasmid, and the resulting plasmid was designated as CAM::TOL*. The introduction of NAH plasmid, belonging to Inc P9 incompatibility group, into P. putida CSTBA carrying CAM::TOLt plasmid and TOL A plasmid did not affect m-toluate catabolism, but resulted in expelling the TOL $\Delta$ plasmid.

• Journal of Microbiology and Biotechnology
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• v.20 no.8
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• pp.1163-1178
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• 2010

Plasmids play important roles in horizontal gene transfer among Vibrionaceae, but surprisingly little is known about their replication and incompatibility systems. In this study, we successfully developed a bioinformatics-assisted strategy of experimental identification of seven Vibrio plasmid replicons. Comparative sequences analysis of the seven Vibrio plasmid replicons obtained in this study together with eight published Vibrionaceae plasmid sequences revealed replication-participating elements involved in the ColE1 mode of replication initiation and regulation. Like plasmid ColE1, these Vibrionaceae plasmids encode two RNA species (the primer RNA and the antisense RNA) for replication initiation and regulation, and as a result, the 15 Vibrionaceae plasmids were designated as ColE1-like Vibrionaceae (CLV) plasmids. Two subgroups were obtained for the 15 CLV plasmids, based on comparison of replicon organization and phylogenetic analysis of replication regions. Coexistence of CLV plasmids were demonstrated by direct sequencing analysis and Southern hybridization, strongly suggesting that the incompatibility of CLV plasmids is determined mainly by the RNA I species like the ColE1-like plasmids. Sequences resembling the conserved Xer recombination sites were also identified on the CLV plasmids, indicating that the CLV plasmids probably use the host site-specific recombination system for multimer resolution like that used by ColE1-like plasmids. All the results indicated that the 15 plasmids form a new ColE1-like group, providing a basis for the rapid characterization and classification of Vibrionaceae plasmids.

• Korean Journal of Veterinary Research
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• v.30 no.2
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• pp.203-214
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• 1990

A total of 166 strains of Salmonella (S) typhimurium var copenhagen isolated from pigeons (164 strains) and aquatic birds (2 strains) were examined for the biochemical characteristics and plasmid profiles. All the strains were sensitive to ampicillin, chloramphenicol, gentamicin, kanamycin and sulfadimethoxine. But 13 strains(7.8%) were resistant to streptomycin (Sm), 2 (1.2%) to tetracycline, 2 (1.2%) to rifampicin, and 1 (0.6%) to nalidixic acid. Among drug resistant strains, only one strain resistant to Sm contained conjugative R plasmid which was fertility inhibition and incompatibility group $I_{\alpha}$. All the strains were sensitive to cobalt chloride, cupric sulfate, lead nitrate, mercuric chloride and silver nitrate. Of 166 isolates, 6 (3.6%) were resistant to sodium arsenate and 1 (0.6%) to potassium tellurite. Among 166 isolates, 1 (0.6%) was colicinogenic, 12 (7.2%) sucrose fermenters, and 166 (100%) maltose fermenters. Plasmid profiles were confirmed as being 4 or 5 plasmids, and their molecular weight ranged 3.2 to 60 megadalton (MD). All the strains harbored 60 Md plasmid. There are three patterns by the plasmid profile, 150 isolates (90.4%) were pattern I (3.2, 3.5, 33, 60Md), 14 (8.4%) pattern II (3.2, 3.5, 29, 60Md), and 2 (1.2%) pattern III (4.2, 7.8, 8.5, 15, 60Md). S typhimurium var copenhagen strains containing 60Md plasmid were resistant to killing by 90% normal guinea pig serum.

• The Journal of the Korean Society for Microbiology
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• v.22 no.1
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• pp.35-53
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• 1987

Multiply resistant Shigella strains isolated in Taegu area were subjected for the characterization of R plasmids. All strains isolated in 1984 and 1985 were susceptible to gentamicin, amikacin, and cephalothin, and most strains were susceptible to kanamycin (Km) and rifampin by agar dilution antimicrobial susceptibility test. The resistance frequency of S. flexneri against ampicillin (Ap) was higher than that of S. sonnei. The strains resistant to sulfisomidine (Su) and trimethoprim (Tp) were found at higher frequency in S. sonnei than in S. flexneri. The most prevalent resistance pattern of S. flexneri was chloramphenicol (Cm) tetracycline (Tc) streptomycin (Sm) Ap, followed by the pattern of CmTcSmSuApTp, CmTcSmSuApTp nalidixic acid, and CmTcSmSuAp in the decreasing order. The antibiogram of CmTcSmSuTp was found to be the most frequent pattern in S. sonnei. The ratio of conjugal transfer of S. flexneri was 47% and 75% of S. sonnei. The average number of plasmid harboring in Shigella was 4 and the size of plasmid ranged 1.3 to 134 megadalton (Mdal). Most S. flexneri carried plasmids of 2 to 3 Mdal and S. sonnei carried those of 3 to 4 Mdal size. The sizes of conjugative plasmids ranged 40-90 Mdal. The incompatibility group (Inc) F II plasmids (54-59 Mdal) were most frequent and rare Inc B plasmids (60 Mdal) of isolates in 1979 and 1980 and Inc FI (87 Mdal) of 1983 isolates were able to be classified by the colony test with standard reference plasmids. The R plasmids of known Inc group were tested for the restriction endonuclease analysis. The pattern of plasmids digested by EcoRl were apparently different by the Inc group but there was no significant difference between species or by the resistance patterns. Nonconjugative plasmids and their phenotypes were identified by transformation test. The transformants were resistant to less than two drugs. Colicin producing transformants carried the Col plasmid of 3.7 or 3.9 Mdal size. $Ap^r$ plasmids derived from S. sonnei were found to be mobilized by transfer factor RT641 to E. coli #CS100. $Ap^r$ plasm ids of same size shared by S. flexneri, S. sonnei, and E. coli were digested with Pstl. All of them showed two restriction fragments of 2.8 kilobase(kb) and 0.7kb. Other plasmids ($Sm^r\;Su^r$) derived from S. flexneri, S. boydii, and S. sonnei were digested with Pstl and they showed same restriction fragment patterns of 3.1kb and 2.9kb. The plasmid profiles of three strains of S. sonnei producing colicin and showing same resistance pattern of CmTcSmSuApTpKm appeared to be similar. Restriction patterns by EcoRl and the behavior of plasmids in conjugation or transformation process were also similar between those plasmids. The restriction patterns were significantly different between the plasmids of Inc FI group and those of unclassified Inc group.