• 한국미생물·생명공학회지
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• 제27권5호
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• pp.364-369
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• 1999

In order to obtain the suitable plasmids for constructing plasmid vectors of Bacillus species, endogeneous plasmid DNAs were screended from thermo-tolerant soil bacteria. Based on agarose gel electrophoresis patterns of the isolated plasmid DNAs, two strains harboring small-size plasmids were selected. The isolated were identified to belong to the genus Bacillus on the basis of their morphological and biochemical properties, and named Bacillus sp. 3-3 and 77-8, respectively. The restriction endonuclease maps were determined for four plasmids including two plasmids from each Bacillus isolates. It is interesting that Bacillus sp. 3-3 and 77-8 have an identical plasmid according to the restriction maps. The three kinds of hybrid plasmids constructed by introducing each plasmid of two isolates into a Escherichia coli plasmid vector. pUCCm18 containing chloramplenicol resistance gene active in Bacillus strains, could be replicated in B. subtilis and B. licheniformis. These plasmids are very stable in B. subtilis, suggesting that the Bacillus plasmids identified in this work would be useful for development of new cloning vectors for Bacillus strains.

• Journal of Microbiology and Biotechnology
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• 제20권8호
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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.

• 미생물학회지
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• 제25권4호
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• pp.255-261
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• 1987

Five independent Actinomycetes harboring plasmids were isolated from soil. Molecular weight of these plasmids was 55kb, 6.2kb, 4.4kb, 55kb and 7.0kb, respectively. Among them small and apprent high copy number plasmids, pJY501 of 4.4kb and pHY711 of 7.0kb, were selected. The plasmids purified by CsCl-EtBr density gradient centrifugation preserved the conformation of supercoiled covalently closed circular molecule, and an apparent copy number was estivated about 150 and about 35 per chromosome. The isolates carrying plasmids were assigned to the genus Streptomyces. For the purpose of introducing selection markers into the isolated plasmids, the tsr fragmemt of pIJ702 was inserted into the BclI site of pJY 501 and pJY711. And the recombinant plasmids constructed designated as pJY502 and pJY712 respectively.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.71.1-71
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• 2003

Most major plant pathogenic bacteria in Korea belong to Xanthomonas spp.. Xanthomonas oryzae pv. oryzae is a major pathogen in rice, X. campestris pv. vesicatoria in pepper, X. axonopodis pv. giycines in soybean, X. campestris pv. campestris in cabbage, and X. axonoposid pv. citri in tangerin. Host specificity of the bacterial pathogen depends on the avirulence gene in the pathogen and the corresponding resistance gene in host plants. Many avirulence genes in bacteiral pathogen located on the native plasmids. However, the presence of the native plasmids in Xanthomonas spp. was not investigated well. In order to study the host specificity, we isolated native plasmids from Xanthomonas spp. and compared those plasmids each other, The presence of the native plasmids and the characteristics of the plasmids depended on the bacterial strains. In the X. axonopodis pv. glycines, most strains carried native plasmids but some strains did not. Some strains carry about 60 kb native plasmids including 3 different aviurlence genes. We will discuss the characteristics of the native plasmids isolated from the Xanthomonas spp.

• 대한수의학회지
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• 제34권4호
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• pp.759-768
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• 1994

E coli strains isolated from pigs were investigated with respect to antimicrobial resistances and prevalence of R-plasmids. Also determined were properties of R-plasmids by plasmid conjugation, curing and southern hybridization using gene probes. All of 400 E coli strains were resistant to CL and SU, and 0.3% to 96.8% of the strains were resistant to most antimicrobials such as TC, PG, AM, SM, CP, GM, EM, NM, etc, while all strains were sensitive to AK. All strains were also multiply resistant to three to twelve antimicrobials. The resistances to PG, SM, TC, AM, CP, SU and ST were transferable and supposed to be mediated by R-plasmids which were opportunistic for transposition into chromosome. Plasmids bigger in size than chromosomal DNA were considered as R-plasmids and most plasmids in small size (<4Kb) proved as cryptic plasmids or nonconjugative R-plasmids. In a strain(No 99), AM resistant property was determined from both chromosomal DNA and R-plasmid DNA which is bigger in size than chromosome.

• 한국어병학회지
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• 제33권2호
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• pp.103-109
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• 2020

In order to use nervous necrosis virus (NNV) virus-like particles (VLPs) as a delivery tool for heterologous antigens or plasmids, we attempted to produce red-spotted grouper nervous necrosis virus (RGNNV) VLPs displaying a partial region of viral hemorrhagic septicemia virus (VHSV) glycoprotein at the surface and VLPs that are harboring DNA vaccine plasmids within the VLP. A peptide encoding 105 amino acids of VHSV glycoprotein was genetically inserted in the loop region of NNV capsid gene, and VLPs expressing the partial part of VHSV glycoprotein were successfully produced. However, in the transmission electron microscope analysis, the shape and size of the partial VHSV glycoprotein-expressing NNV VLPs were irregular and variable, respectively, indicating that the normal assembly of capsid proteins was inhibited by the relatively long foreign peptide (105 aa) on the loop region. To encapsulate by simultaneous transformation with both NNV capsid gene expressing plasmids and DNA vaccine plasmids (having an eGFP expressing cassette under the CMV promoter), NNV VLPs containing plasmids were produced. The encapsulation of plasmids in the NNV VLPs was demonstrated by PCR and cells exposed to the VLPs encapsulating DNA vaccine plasmids showed fluorescence. These results suggest that the encapsulation of plasmids in NNV VLPs can be done with a simple one-step process, excluding the process of disassembly-reassembly of VLPs, and NNV VLPs can be used as a delivery tool for DNA vaccine vectors.

• 대한미생물학회지
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• 제22권1호
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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.

• 대한수의학회지
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• 제29권2호
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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.

• The Korean Journal of Ecology
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• 제11권1호
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• pp.29-36
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• 1988

This present study has been carried out to examine the correlation between the distribution of the coliform bacterial plasmids and the viability test against heavy metals in the upper stream of Han river(Gapyung; clean water) and the its basin (Anyang Cheon; polluted water). And the distribution of plasmids were examined to be used as trophic indicator for analysis between the clean and polluted waters. 1. A total of 110 isolates were analyzed for the presence of plasmids by means of the boiling method and agarose gel techniques. Plasmids were significantly more frequent in the strains which had been isolated from the clean water (14.3%). Also, there were much higher multiplicity of plasmids at the polluted water(41.0%), compared with the clean water (33.0%). By the comparision between molecular weight of bacterial plasmids in the clean water and those of polluted water, there were no significant differences of the clean water from the polluted water, to such extent as 30.0%, 28.6%, respectively, in frequency for occurrence of high molecular weight plasmids iver 35.8% Mdal. 2. Each isolate was carried out the resistance test for mercury(10-5M), nickel(10-3M) and arsenic (0.1%). At the polluted site, the survival ability of the plasmid-carrying straings(Hg, 31.0%l Ni, 5.7%; As, 65.7%) was higher than that of the non plasmid-carrying strains(Hg, 12.1%; Ni, 3.0%; As, 54.6%). This trend was more remarkable in the clean site (plasmid-carrying strains: 16.7%,-,-). As a result it is suggested that plasmids could be used as an indicator of a certain types of water pollution. In addition, heavy metals might have inflyenced, some extent, to the distribution of plasmids in the environment which has been surveyed in the present study.

• 대한미생물학회지
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• 제22권3호
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• pp.267-273
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• 1987

In the analysis of plasmid profile obtained by agarose gel electrophoresis, the strain harvoring multiple reference plasmids of known molecular weight were needed to estimate the size of unknown molecular weight plasmids. Six strains of E. coli isolated from clinical specimens carried multiple plasmids and these strains could be available as a reference plasmids harvoring strain. These E. coli strains showed 4 to 9 plasmids of various size ranging 2.5 to 94.3 megadalton (Mdal). The correlation coefficients of linear regression between the relative mobility and molecular weight were 0.99966 to 1.00000. Among them, E. coli KE327 from throat which contained 7 plasm ids as follows: 79 Mdal, 46 Mdal, 33 Mdal(pKY3027 C), 4.9Mdal, 3.8Mdal(pKY3027 E), 3.5Mdal, and 2.7 Mdal. Relative amount of the pKY3027 C was the smallest(7.09%) and that of the pKY3027 E was the largest (24.24%) among the plasmid fractions of E. coli KE327.