• Title, Summary, Keyword: plasmids

Search Result 588, Processing Time 0.042 seconds

Characterization of Endogeneous Plasmids from Two Bacillus Isolates (Bacillus 속 분리균 2종의 내재형 Plasmids 특성분석)

  • 윤기홍
    • Microbiology and Biotechnology Letters
    • /
    • v.27 no.5
    • /
    • pp.364-369
    • /
    • 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.

  • PDF

A New ColE1-like Plasmid Group Revealed by Comparative Analysis of the Replication Proficient Fragments of Vibrionaceae Plasmids

  • Pan, Li;Leung, P.C.;Gu, Ji-Dong
    • Journal of Microbiology and Biotechnology
    • /
    • v.20 no.8
    • /
    • pp.1163-1178
    • /
    • 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.

Isolation and characterization of plasmids isolated from streptomyces spp. and construction of recombinant plasmids (Streptomyces 속으로 부터 분리한 플라스미드의 특성 및 재조합 유도체의 제조)

  • 유주현;염도영;공인수
    • Korean Journal of Microbiology
    • /
    • v.25 no.4
    • /
    • pp.255-261
    • /
    • 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.

  • PDF

Isolation and characterization of native plasmids carrying avirulence genes in Xanthomonas spp.

  • Sunggi hen;Lee, Seungdon;Jaewoong Jee;Park, Minsun
    • Proceedings of the Korean Society of Plant Pathology Conference
    • /
    • /
    • pp.71.1-71
    • /
    • 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.

  • PDF

Distribution of antimicrobial resistances and properties of R-plasmids in E coli isolated from pigs (돼지유래 대장균의 항균제내성 분포와 R-plasmid의 성상)

  • Chung, Myeong-eun;Yeo, Sang-geon
    • Korean Journal of Veterinary Research
    • /
    • v.34 no.4
    • /
    • pp.759-768
    • /
    • 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.

  • PDF

Molecular Characteristics of R Plasmids in Shigella (Shigella R Plasmid의 분자적 특성)

  • Lee, Yoo-Chul;Seol, Sung-Yong;Cho, Dong-Taek;Chun, Do-Ki
    • The Journal of the Korean Society for Microbiology
    • /
    • v.22 no.1
    • /
    • pp.35-53
    • /
    • 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.

  • PDF

Dynamics of Supercoiled and Relaxed pTZ18U Plasmids Probed with a Long-Lifetime Metal-Ligand Complex

  • Kang, Jung-Sook;Abugo, Omoefe O.;Lakowicz, Joseph R.
    • BMB Reports
    • /
    • v.35 no.4
    • /
    • pp.389-394
    • /
    • 2002
  • $[Ru(bpy)_2(dppz)]^2+$ (bpy=2,2'-bipyfidine, dppz=dipyrido[3,2-a:2',3'-c]phenazine) (RuBD), a long-lifetime metal-ligand complex, displays favorable photophysical properties. These include long lifetime, polarized emission, but no significant fluorescence from the complex that is not bound to DNA. To show the usefulness of this luminophore (RuBD) for probing the bending and torsional dynamics of nucleic acids, its intensity and anisotropy decays when intercalated into supercoiled and relaxed pTZ18U plasmids were examined using frequency-domain fluorometry with a blue light-emitting diode (LED) as the modulated light source. The mean lifetimes for the supercoiled plasmids (< $\tau$ >=148 ns) were somewhat shorter than those for the relaxed plasmids (< $\tau$ >=160 ns). This suggests that the relaxed plasmids were shielded more efficiently from water. The anisotropy decay data also showed somewhat shorter slow rotational correlation times for supercoiled plasmids (288 ns) than for the relaxed plasmids (355 ns). The presence of two rotational correlation times suggests that RuBD reveals both the bending and torsional motions of the plasmids. These results indicate that RuBD can be useful for studying both the bending and torsional dynamics of mucleic acids.

Genentic properties of R plasmids in Salmonella isolates of swine and bovine origin in Korea II. Incompatibility and profile of R plasmid (우(牛), 돈(豚)에서 분리(分離)한 Salmonella유래(由來) R plasmid의 유전학적(遺傳學的) 및 분자생물학적(分子生物學的) 성상(性狀)에 관한 연구(硏究) II. R plasmid의 비적합성(非適合性) 및 plasmid profile)

  • Choi, Won-pil;Lee, Hi-suk;Yeo, San-geon;Lee, Hun-jun;Jung, Suk-chan
    • Korean Journal of Veterinary Research
    • /
    • v.29 no.2
    • /
    • pp.59-67
    • /
    • 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.

  • PDF

Stability of Recombinant Plasmids Carrying the stb Locus of E. coli IncFII NR1 Plasmid in E. coli and Yeast (대장균과 효모에서 Escherichia coli IncFII NR1 플라스미드의 stb 좌위를 포함하는 재조합 플라스미드의 안정성에 관한 연구)

  • Chung, Kung-Sook;Kim, Choon-Kwang;Kim, Kyu-Won
    • Korean Journal of Microbiology
    • /
    • v.31 no.1
    • /
    • pp.37-43
    • /
    • 1993
  • The effect of stb locus of E. COLI IncFII plasmid NR1 on the stability of chimeric plasmids was investigated. First, we have isolated the stability locus (stb) from E. coli NR1 plasmid and then inserted into the three different vectors, pUC8, YRp17 and YEp24. By examining their stability in E. coli and yeast, we showed that the recombinant plasmids containing stb locus were resonably stable. Also, by comparing the amounts of the rDNA fragments per haploid genome with those of the plasmid fragments, we showed they copy number of recombinant plasmids was not increased. Consequently, the stb locus of E. coli IncFII plasmid NR1 stabilized the chimeric plasmids but did not affect the replication or copy number of plasmids.

  • PDF

Application of Coliform Bacterial Plasmid as a Trophic Indicator (콜리형 세균 Plasmid의 영양단계 지표 활용에 관한 연구)

  • Cho, Hong Bum
    • The Korean Journal of Ecology
    • /
    • v.11 no.1
    • /
    • pp.29-36
    • /
    • 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.

  • PDF