• Title/Summary/Keyword: multiple plasmids

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The Use of the Strain Containing Multiple Plasmids as Size Reference Plasmids (분자량측정을 위한 Reference Plasmid 보유균주의 이용)

  • Bang, Sung-Hyuk;Lee, Yoo-Chul;Seol, Sung-Yong;Cho, Dong-Taek
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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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.

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Curing Both Virulent Mega-Plasmids from Bacillus anthracis Wild-Type Strain A16 Simultaneously Using Plasmid Incompatibility

  • Wang, Dongshu;Gao, Zhiqi;Wang, Huagui;Feng, Erling;Zhu, Li;Liu, Xiankai;Wang, Hengliang
    • 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.

Characterization of Plasmids from Multiple Antibiotic Resistant Vibrio sp. Isolated from Molluscs and Crustaceans

  • Manjusha, Sayd;Sarita, Ganabhat Bhat
    • Microbiology and Biotechnology Letters
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    • v.40 no.3
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    • pp.197-207
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    • 2012
  • This study investigated the role of plasmids and their relationship with the multiple antibiotic resistance of 30 Vibrios sp. isolated from molluscs and crustaceans sampled from the Kerala coastal waters of India. The biochemical identification and antibiotic resistance profiles were determined, followed by the plasmid profiles, conjugation and transformation efficiencies. The results showed a considerable difference in the level of bacterial resistance to various antibiotics; while all 30 strains were found to be MAR Vibrios sp. and their resistance patterns varied. All the strains were resistant to amoxycillin, ampicillin and carbeniciliin. 87% were resistant to rifampicin; 74% to cefuroxime; 67 to streptomycin; 53% to norfloxacin and ciprofloxacin and 47% to furazolidone and nalidixic acid. In addition to their antibiotic resistance, the plasmid DNA of the MAR Vibrios strains isolated from the molluscs and crustaceans was also studied. Nine strains isolated from crustaceans and molluscs were found to harbor 1-3 plasmids with sizes varying from 5. 98 kb to 19. 36 kb. The average transformation efficiency was about $5{\times}10^{-8}$ and the conjugation efficiency varied from $2.1{\times}10^{-3}$ to $10^{-9}$. A further study of antibiotic resistance patterns may be useful to test the extent of drug resistance in seafoods and help to devise a nationwide antibiotic policy.

R-plasmids in staphylococcus aureus (Staphylococcus aureus의 항생제 내성 plasmid에 관한 연구)

  • 변우현;김영선;조은희;권동현;이호주;홍순주
    • Korean Journal of Microbiology
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    • v.23 no.4
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    • pp.282-290
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    • 1985
  • Small size antibiotic resistance plasmids having molecular weights less than 10 Mdal were isolated and characterized from ten clinically isolated multiple resistant Staphylococcus aureus. Agarose gel electrophoresis profiles and antibiotic resistance patterns divided these strains into four groups. Strain 2-23-6, the representative strain of a group of five strains conferred two plasmids of molecular weights $1.6{\times}10^6\;dal\;and\;2.0{\times}10^6$ dal. The small plasmid (pSBK 112) specified macrolides, lincosamides and streptogramin type B (MLS) resistance gene which are expressed constitutively. Lage plasmid (pSBK 125) specified chloramphenicol resistance gene which is inducible. Strain 10-5 conferred a $3.0{\times}10^6$ dal plasmid (pSBK 141) which carry an inducible ampicillin resistance gene and strain P-H-2 conferred and $1.6{\times}10^6$ dal plasmid (pSBK 190) which carry a constitutive MLS resistance gene. Strain D-H-1 conferred four plasmids of molecular weights $0.8{\times}10^6$ dal (pSBK 201), $1.6{\times}10^6$ dal (pSBK 202), $2.5{\times}10^6$ dal (pSBK 203), and $1.2{\times}10^7$ dal (pDBK 204), respectively. Among those four plasmids, only pSBK 203 specified chloramphenicol resistance gene. Curing of constitutive MLS resistance using acriding orange or ethidium bromide in 2-23-6 and P-H-2 strains produced 'inducible' MLS resistance strains which are less resistant to MLS than the wild type strains, suggesting that there are two resistance genes in both strains; one is constitutive and the other is inducible.

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Characterization of Plasmids of Chloramphenicol Resistant Staphylococcus aureus (클로람페니콜 내성 황색포도상구균의 플라스미드 동정)

  • Park, Jung-Hee;Lee, Jae-Yoon;Moon, Kyung-Ho
    • YAKHAK HOEJI
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    • v.52 no.1
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    • pp.33-36
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    • 2008
  • Staphylococcus aureus KH13 and Staphylococcus aureus KH28 were resistant to chloramphenicol, ampicillin, clindamycin, erythromycin, gentamicin, kanamycin, streptomycin, tobramycin, and norfloxacin. A plasmid (pKH13) and two plasmids (pKH14, pKH15) were isolated from Staphylococcus aureus KH13 and Staphylococcus aureus KH28, respectively and complete nucleotide sequences of three plasmids were determined. It was found that pKH13 and pKH15 mediated chloramphenicol resistance and pKH14 was a cryptic plasmid.

Complete genome sequence of multidrug-resistant Moraxella osloensis NP7 with multiple plasmids isolated from human skin (사람의 피부에서 분리한 다약제 내성이며 다수의 플라스미드를 갖는 Moraxella osloensis NP7 균주의 유전체 서열 분석)

  • Ganzorig, Munkhtsatsral;Lim, Jae Yun;Hwang, Ingyu;Lee, Kyoung
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.286-288
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    • 2018
  • Moraxella osloensis NP7 was isolated from human skin of a collage male and showed resistance to ${\beta}-lactam$ and aminoglycoside antibiotics. Herein, we report the complete whole-genome sequence and gene annotations of M. osloensis NP7. It possesses single circular chromosome and seven plasmids. Chromosome is 2,389,582 bp in length with the G + C content of 43.9% and encodes 2,065 protein-coding genes. The combined seven plasmids are 654,202 bp in size with the average G + C content of 40.5% and code for a total of 667 protein-coding genes. The chromosome of NP7 strain contains four ribosomal RNA operon copies, one transfer-messenger RNA gene, forty-seven tRNA genes, three riboswitch genes and three CRISPR arrays. Additional CRISPR array is found in the plasmid pNP7-1. The genes conferring resistance to ${\beta}-lactam$ and aminoglycoside antibiotics were predicted to reside in the plasmid pNP7-1.

Incidence of Plasmids in Marine Bacteria Isolated from the Bunker-C Oil Enriched Culture (Bunker-C유 집식배양으로부터 해양세균 Plasmid의 분포)

  • Park, In-Sick;Park, Jung-Youn;Jin, Deuk-Hee;Hong, Yong-Ki
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.270-274
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    • 1988
  • Samples used for the enrichment culture were collected from the sea water of suspected chronic petroleum contamination in the vicinity of Pusan, Chungmu and Ulsan ports, Korea. Alkaline Iysis and agarose gel electrophoresis techniques were employed to screen these isolates for the presence of plasmid DNA. There were n little differences in the percentage of isolates containing plasmids between sampling sites of unpolluted sen water (22%) and polluted son water (25%). Bacterial isolates taken from the Bunker-C oil enriched culture showed significantly more plasmid incidence (29%). About two thirds of strains grown on a variety of hydrocarbons were Gram negative strains of which 33% contained one or more plasmids. Multiple plasmids were observed in 23% of the plasmid-carrying strains. Forty one percent of the plasmids detected were estimated to have a mass of 20 kb or more.

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Antibiotic Resistance and R Plasmids in Edwardsiella tarda (양만장 사육조에서 분리한 Edwardsiella tarda의 약제 내성과 R Plasmid)

  • Choi, Min-Soon;Kim, Young-Gill
    • Journal of fish pathology
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    • v.7 no.1
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    • pp.37-46
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    • 1994
  • A total 103 strains of Edwardsiella tarda was isolated from eel culture-ponds and examined for drug resistance, distribution and transferabilities of R plasmids. The drugs used were lincomycin(LM), penicillin(PM), sulfamethazine(SF), sulfadimethoxine(SD), cephalosprin(CP), chloramphenicol(CH), streptomycin(SM), oxytetracycline(OT), ampicillin(AP), oxolinic acid(OA), kanamycin(KM), amikacin(AK), gentamicin(GM) and enrofloxacin(EF). Two strains were resistant to the all drugs used, and all isolates were multiply resistant to drugs(at least 8 among 14 drugs), mostly restricted to LM(103 strains), PM(103 strains), SD(103 strains), SF(103 strains), CP(102 strains), CM(101 strains), SM(100 strains), OT(94 strains), AM(92 strains), OA(80 strains), KM(60 strains), AK(30 strains), GM(19 strains) and EF(14 strains), in combination at high degree showing 34 different drug resistant patterns. The most frequently encountered patterns were LM PM SD SF CP CH SM OT AP OA KM(22 strains, 22.4%) followed by LM PM SD SF CP CH SM OT AP OT(12 strains, 11.7%). LM PM SD SF CP CH SM OT AP(7 strains, 6.8%), LM PM SD SF CP CH SM OT AP OA KM AK GM(6 strains, 5.8%) and LM PM SD SF CP CH SM OT AM OX KM AK GM(6 strains, 5.8%). Transfer experiment of drug resistance showed that of 103 resistant strains, 100 strains(97%) transferred part or all resistance to all drugs, indicating that most isolates carried conjugally transferrable R plasmids determining multiple drugs. The most frequently observed transferarble R plasmids were LM PM SD SF CP CH SM OT AP OA(10 strains), LM PM SD SF CP CH SM OT AP OX KM(7 strains) and LM PM SD SF CP CH AP OA(7 strains). These results sugested that eel culture-ponds were highly contaminated with different strains of Edwardsiella tarda, and that contaminated bacteria might be highly multiple resistant strains to drugs, carring transferable R plasmids.

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Simplex PCR Assay for Detection of blaTEM and gyrA Genes, Antimicrobial Susceptibility Pattern and Plasmid Profile of Salmonella spp. Isolated from Stool and Raw Meat Samples in Niger State, Nigeria

  • Musa, Dickson A.;Aremu, Kolawole H.;Ajayi, Abraham;Smith, Stella I.
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.230-235
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    • 2020
  • The global evolution of antibiotic resistance has threatened the efficacy of available treatment options with ravaging impacts observed in developing countries. As a result, investigations into the prevalence of antibiotic resistance and the role of plasmids are crucial. In this study, we investigated the presence and distribution of blaTEM and gyrA genes, plasmid profiles, and the antimicrobial susceptibility pattern of Salmonella strains isolated from raw meat and stool sources across Niger State, Nigeria. Ninety-eight samples, comprising 72 raw meat and 26 stool samples, were screened for Salmonella spp. The antimicrobial susceptibility of Salmonella isolates to 10 commonly used antimicrobial agents was determined using the KirbyBauer disc diffusion method. Isolates were further analyzed for plasmids, in addition to PCR amplification of beta-lactamase (blaTEM) and gyrA genes. A total of 31 Salmonella spp. were isolated, with 22 from raw meat (70.97%) and 9 from stool (29.03%). Salmonella spp. with multiple resistance patterns to ceftazidime, cefuroxime, ceftriaxone, erythromycin, ampicillin, cloxacillin, and gentamicin were detected. Ofloxacin and ciprofloxacin were found to be the most effective among the antibiotics tested, with 67.7% and 93.5% susceptible isolates, respectively. Nine (29.03%) isolates harbored plasmids with molecular sizes ranging between 6557 bp and 23137 bp. PCR amplification of gyrA was detected in 1 (3.23%) of the 31 isolates while 28 isolates (90.32%) were positive for blaTEM. This study shows the incidence of antibiotic resistance in Salmonella isolates and the possible role of plasmids; it also highlights the prevalence of ampicillin resistance in this local population.

Rhizosphere Communication: Quorum Sensing by the Rhizobia

  • He, Xuesong;Fuqua, Clay
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1661-1677
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    • 2006
  • Rhizobium and related genera are soil bacteria with great metabolic plasticity. These microorganisms survive in many different environments and are capable of eliciting the formation of nitrogen-fixing nodules on legumes. The successful establishment of symbiosis is precisely regulated and requires a series of signal exchanges between the two partners. Quorum sensing (QS) is a prevalent form of population density-dependent gene regulation. Recently, increasing evidence indicates that rhizobial quorum sensing provides a pervasive regulatory network, which plays a more generalized role in the physiological activity of free-living rhizobia, as well as during symbiosis. Several rhizobia utilize multiple, overlapping quorum sensing systems to regulate diverse properties, including conjugal transfer and copy number control of plasmids, exopolysaccharide biosynthesis, rhizosphere-related functions, and cell growth. Genomic and proteomic analyses have begun to reveal the wide range of functions under quorum-sensing control.