• 제목/요약/키워드: multiple plasmids

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

  • 방성혁;이유철;설성용;조동택
    • 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.

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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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    • 제25권10호
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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
    • 한국미생물·생명공학회지
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    • 제40권3호
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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.

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

  • 변우현;김영선;조은희;권동현;이호주;홍순주
    • 미생물학회지
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    • 제23권4호
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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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클로람페니콜 내성 황색포도상구균의 플라스미드 동정 (Characterization of Plasmids of Chloramphenicol Resistant Staphylococcus aureus)

  • 박정희;이재윤;문경호
    • 약학회지
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    • 제52권1호
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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.

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

  • 간조리그 뭉크사츠랄;임재윤;황인규;이경
    • 미생물학회지
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    • 제54권3호
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    • pp.286-288
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    • 2018
  • 남자 대학생의 피부에서 분리한 Moraxella osloensis NP7는 베타-락탐과 아미노글리코사이드 항생제에 대해 내성을 보였다. 본 연구에서는 NP7 균주 유전체의 완전한 염기서열과 유전자 주석을 보고하고자 한다. NP7 균주는 원형 염색체와 7개의 플라스미드를 갖고 있다. 염색체는 43.9%의 G + C 함량을 갖는 2,389,582개의 염기쌍을 갖고 있으며, 단백질을 암호하는 2,065개의 유전자를 보유하고 있다. 전체 플라스미드는 평균적으로 40.5%의 G + C 함량을 갖는 654,202개의 염기쌍을 갖고 있으며, 단백질을 암호하는 667개의 유전자를 보유하고 있다. 염색체는 4개의 리보좀 RNA 오페론, 1개의 transfermessenger RNA 유전자, 47개의 tRNA 유전자, 3개의 핵산스위치 유전자 그리고 3개의CRISPR array를 포함하고 있으며, 1개의 CRISPR은 pNP7-1 플라스미드에 존재한다. 베타-락탐과 아미노글리코사이드 항생제에 내성을 부여하는 유전자는 pNP7-1 플라스미드에 존재하고 있다.

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

  • Park, In-Sick;Park, Jung-Youn;Jin, Deuk-Hee;Hong, Yong-Ki
    • 한국미생물·생명공학회지
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    • 제16권4호
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    • pp.270-274
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    • 1988
  • 우리나라 연근해역의 해양오염중 주종을 이루는 Bunker-C유를 대상으로 부산 및 충무, 울산항구의 해수를 균원으로 접종하여 enrichment culture시켰다. 이와 같은 혼합배양 해양세균들의 plasmid DNA분포를 agarose gel 전기영동상에서 조사하여 보았다. 우선 오염되지 않은 해수로부터 분리한 세균들의 Plasmid 분포(22%)와 만성적으로 유류오염이 예상되는 각 항구의 세균 plasmid 분포(25%)에는 조금 차이는 있었으며 enrichment culture중에는 29%정도로서 plasmid 분포비율이 증가하였다. 각종 탄화수소 화합물들에 성장하는 세균중 약33%는 plasmid룰 함유하고 있었으며, 또 약 62%정도는 Gram 음성균이었다. Plasmid 함유 세균중 23% 정도는 2종 이상의 plasmid를 갖고 있으며, 41%의 plasmid는 20kb 이상의 크기를 가지고 있는 것으로 나타났다.

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

  • 최민순;김영길
    • 한국어병학회지
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    • 제7권1호
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    • pp.37-46
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    • 1994
  • 서해안 일원의 가온 양만장 사육조로부터 분리한 E. tarda균에 대한 약제내성검사 및 내성균의 내성인자 전달 시험을 실시하였다. 공시한 항생제에 대한 내성률은 LM, PM, SD, SF 등에는 100%, AP, CH, SM, OT 및 CP 등에는 89% 이상, OA 및 KM에는 70% 정도 그리고 AK, GM 및 EF 등에는 30% 이하의 내성률을 보였으며, 이와같은 내성률은 각 양만장에서 분리한 균주간에 차이를 보이지 않았다. 내성유형은 총 34유형으로 모든 분리균주가 8~14제의 다제내성을 보였으며, 그 중 LM PM SD SF CP CH SM OT AP OA KM(22주, 21.4%) 및 LM PM SD SF CP CH SM OT AP OA(12주, 11.7%) 유형등이 고빈도로 출현하였다. 내성균의 내성인자 전달유형은 총 55형으로 단제에서 13제까지의 다양한 내성인자가 전달되었는데, 그 중 LM PM SD SF CP CH SM OT AP OA(10주. 10%), LM PM SD SF CP CH SM OT AP OA KM(7주, 7%) 및 LM PM SD SF CP CH AP OA(7주, 7%) 등의 다제내성유형이 고빈도의 전달률을 보였으며, 약재별로는 GM, EF 및 AK 등에는 저빈도(0~20%), OT, SM, KM 및 OA 등에는 중등도(49~62%), AP, PM, LM, SF, CP, CH 및 SD 등에는 고빈도(71~94%)로 내성인자를 전달하였다.

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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.
    • 한국미생물·생명공학회지
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    • 제48권2호
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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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    • 제16권11호
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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.