• Title/Summary/Keyword: R plasmids

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$Ginsenoside-R_{b1}$ Acts as a Weak Phytoestrogen in MCF-7 Human Breast Cancer Cells

  • Lee, Young-Joo;Jin, Young-Ran;Lim, Won-Chung;Park, Wan-Kyu;Cho, Jung-Yoon;Jang, Si-Youl;Lee, Seung-Ki
    • Archives of Pharmacal Research
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    • v.26 no.1
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    • pp.58-63
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    • 2003
  • Ginseng has been recommended to alleviate the menopausal symptoms, which indicates that components of ginseng very likely contain estrogenic activity. We have examined the possibility that a component of Panax ginseng, $ginsenoside-R_{b1}$ acts by binding to estrogen receptor. We have investigated the estrogenic activity of $ginsenoside-R_{b1}$ in a transient transfection system using estrogen-responsive luciferase plasmids in MCF-7 cells. $ginsenoside-R_{b1}$ activated the transcription of the estrogen-responsive luciferase reporter gene in MCF-7 breast cancer cells at a concentration of 50 $\mu$M. Activation was inhibited by the specific estrogen receptor antagonist ICI 182,780, indicating that the estrogenic effect of $ginsenoside-R_{b1}$ is estrogen receptor dependent. Next, we evaluated the ability of $ginsenoside-R_{b1}$ to induce the estrogen-responsive gene c-fos by semi-quantitative RT-PCR assays and Western analyses. $ginsenoside-R_{b1}$ increased c-fos both at mRNA and protein levels. However, $ginsenoside-R_{b1}$ failed to activate the glucocorticoid receptor, the retinoic acid receptor, or the androgen receptor in CV-1 cells transiently transfected with the corresponding steroid hormone receptors and hormone responsive reporter plasmids. These data support our hypothesis that $ginsenoside-R_{b1}$ acts a weak phytoestrogen, presumably by binding and activating the estrogen receptor.

Genetic properties of R plasmids in Salmonella isolates of swine and bovine origin in Korea I. Distribution and drug resistance of Salmonella isolated from dairy cow (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) I. 유우(乳牛)에서 Salmonella속균(屬菌)의 분포상황(分布狀況) 및 약제내성(藥劑耐性))

  • Choi, Won-pil;Lee, Hi-suk;Yeo, San-geon;Lee, Hun-jun;Chae, Tae-chul
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.331-337
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    • 1988
  • This paper dealt with the distribution of Salmonella (S) infection on 4 herds in Kyungju and Taegu during the period from May to October 1986. Isolated Salmonella were examined for serotypes, antimicrobial drug resistance and detection of R plasmid. The results obtained were summarised as followings: 1. Of total 4.622 samples from 4 herds, 67 Salmonella were isolated from 51 samples(1.1%), and their serovar strains were S typhimurium 6, S derby 5, S infantis 4, S bareilly 4, S dublin 3, S anatum 2, S montevideo 2 and untypable 41. 2. The isolation rate of Salmonella was higher in summer and autumn. 3. Of the 67 strains examined, 45 (67.2%) were resistant to one or more antibiotics, such as ampicillin (Am), cephalothin (Ce), chloramphenicol (Cm), rifampicin (Rf), sulfadimethoxine (Su), and tetracycline (Tc), and higher resistant to Sm (40.2%), Ce (31.3%), Am (23.9%). 4. Of the 45 resistant Salmonella strains, 44 (97.8%) harbored conjugative R plasmids and the transfer frequency of Sm (100%), Ce (95.2%), Tc (91.0%) and Su (80.0%) resistance was much higher than that of the other drug resistance. 5. The most common resistant patterns were Sm, Ce, AmCeCmSmSuTc, and AmCe. 6. In 4 herds, the incidience of drug resistance was 57.7%~100% and transfer frequency of conjugative R plasmid was 96.1%~100%.

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Characteristics of MCPA plasmid isolated from pseudomonas sp. (Pseudomonas에서 분리한 MCPA 플라스미드의 특성)

  • 이영록;최대성;은성호;박영두
    • Korean Journal of Microbiology
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    • v.24 no.4
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    • pp.394-399
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    • 1986
  • From the lysates of the 7 selected strains of Pseulomonas utilizing 2-methyl-4-chlorophenoxyactate as a sole source of carbon and energy, several MCPA plasmids, which encodes genes for the degradation of 2-methyl-4-chlorophenoxyacetate, were isolated, and measured their molecular weight as well as genetic characters such as resistance to antibiotics and degradative ability of other chlorinated herbicides. Transmissibility of the MCPA plasmids, pKU1, pKU15, and pKU17 was tested by conjugation or transformation and the restriction pattern of pKU15 for Pvu II, Hind III, EcoR I, Xho I, Bgl II, and Ava II was analyzed.

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Enhancement of Clavulanic Acid by Replicative and Integrative Expression of ccaR and cas2 in Streptomyces clavuligerus NRRL3585

  • Hung, Trinh Viet;Malla, Sailesh;Park, Byoung-Chul;Liou, Kwang-Kyoung;Lee, Hei-Chan;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1538-1545
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    • 2007
  • Clavulanic acid (CA) is an inhibitor of ${\beta}$-lactamase that is produced from Streptomyces clavuligerus NRRL3585 and is used in combination with other antibiotics in clinical treatments. In order to increase the production of CA, the replicative and integrative expressions of ccaR (encoding for a specific regulator of the CA biosynthetic operon) and cas2 (encoding for the rate-limiting enzyme in the CA biosynthetic pathway) were applied. Six recombinant plasmids were designed for this study. The pIBRHL1, pIBRHL3, and pIBRHL13 were constructed for overexpression, whereas pNQ3, pNQ2, and pNQ1 were constructed for chromosomal integration with ccaR, cas2, and ccaR-cas2, respectively. All of these plasmids were transformed into S. clavuligerus NRRL3585. CA production in transformants resulted in a significantly enhanced amount greater than that of the wild type, a 2.25-fold increase with pIBRHLl, a 9.28-fold increase with pNQ3, a 5.06-fold increase with pIBRHL3, a 2.93-fold increase with pNQ2 integration, a 5.79-fold increase with pIBRHLl3, and a 23.8-fold increase with pNQ1. The integrative pNQl strain has been successfully applied to enhance production.

Prevalence and Drug Resistance of Shigella in Taegu Area of Korea (대구지방에서 분리된 Shigella의 양상과 항균제 내성)

  • Chun, Do-Ki;Park, Jong-Wook;Suh, Seong-Il;Cho, Dong-Taek;Seol, Sung-Yong;Lee, Yoo-Chul
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.461-471
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    • 1986
  • Shigella strains isloated in the Teagu area during the period from 1973 to 1985 were studied for species distribution, drug resistance, and R plasmids. Approximately 1,200 strains were isolated during this period, and most of them were classified into Shigella flexneri, S. sonnei occupied less than 20%, and S. dysenteriae and S. boydii were very rarely isolated. More than 95% of them were resistant to one or more of these drugs; chloramphenicol (Cm), tetracycline (Tc), streptomycin (Sm), sulfisomidine (Su), ampicillin (Ap), and trimethoprim (Tp). Strains resistant to kanamycin, nalidixic acid (Na), and rifampin (Rf) were rare, and no strain was resistant to cephaloridine, gentamicin, and amikacin. Approximately half of the isolates were resistant to drugs in 1973, but the rate of resistant strains increased to more than 95% from 1977. Strains resistant to the four drugs (Cm, Tc, Sm, and Su) occupied the majority of resistant strains until 1977, but the most prevalent multiplicity of drug resistance increased to six drugs (Cm, Tc, Sm, Su, Ap, and Tp) from 1978 with the marked increase of Ap- and Tp-resistant strains. Approximately 75% of them transferred resistance to Escherichia coli by conjugation, and the resistance was considered to be mediated by R plasmids. Almost all of them transferred the complete patterns of resistance to drugs except Na and Rf. However, among some strains of recent isolates, small numbers of segregants of transferred resistance were observed. The R plasmids in Shigella were mostly classified into Inc FII, and only small numbers into Inc B. Segregants were in most cases unclassified.

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Survey of drug resistance in Edwardsiella tarda isolated from diseased eels(Anguilla japonica) (뱀장어 병어로부터 분리한 Edwardsiella tarda의 약제내성)

  • Choi, Min-Soon;Choi, Sang-Hoon;Park, Kwan-Ha;Jang, Seon-Il;Yoon, Chang-Yong;Cho, Jeong-Gon;Song, Hee-Jong
    • Journal of fish pathology
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    • v.9 no.2
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    • pp.195-201
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    • 1996
  • Ninety-six isolates of Edwardsiella tarda recovered from outbreaks of Edwardsiellosis in cultured eels(Anguilla japonica) in Kunsan, were examined for drug susceptibility, distribution and transferabilities of R plasmid. All of the E. tarda isolates examined were sensitive to gentamicin(GM), streptomycin(SM), norfloxacin(NF), and amikacin(AK). But most isolates were resistant to sulfadimethoxine(SD, 86 strains), ampicillin(AM, 84 strains), penicillin G(PM, 80 strains), nalidixic acid (NA, 67 strains), oxytetracycline(OT, 44 strains), and oxolinic acid(OA, 37 strains). Twenty different combinations of drug resistance patterns were observed : the frequently encountered pattern was SD-AM-PM-NA-OA(16 strains), SD-AM-PM-NA(14 strains), SD-AM-PM-NA-OT-OA(12 strains), SD-AM-PM-OT(10 strains), and SD-AM-PM-NA-OT(8 strains). Transferable R plasmids were found out to be carried in 78 out of 94 resistant strains, indicating that these isolates carry conjugally transferable R plasmids associated with single or multiple drugs. The frequently observed transferarble R plasmids were AM(8 strains), AM-PM-NA(8 strains), Am-SD(6 strains), PM(6 strains), and SD(6 strains) These results suggest that high dose of various antibacterials might have already been introduced to eel culture system leading to the acquirement of multi-drug resistance to wide range of antibacterials.

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유전공학적으로 변형시킨 R-plasmid 들의 전이에 미치는 균주와 pH 의 영향

  • 김희태;이성기;김치경
    • Korean Journal of Microbiology
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    • v.30 no.2
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    • pp.88-95
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    • 1992
  • The genetically engineered microorganisms (GEMS) could be released accidentally or ii)rexperimental purposes, as the genetic engineering, technique ha:. become very popular inany laboratories of biological sciences. But there have been littlt: informations on transkrbehavior of the genetically ~nodified genes in the natural en\ironmentx. In this stutly.antibiotic resistant bacteria were isolated from nat.ural waters. and then GEM strains wereconstructed i'rom the natural isolate (NI) by ~noclification oi' the Km' plasmitl. Thetransferability of the plasmids in the GEM and NI strains were examinetl by con-jugationin Luria-Bertani broth :it 30$^{\circ}$C. Also the cff'ccts 01' mating strain and pH on their transferfrequency and rearrangement of the plasmids in tl-~ec o~ijugantsM ere comp:irati\ely stuclictl.I'hc transkr frequency of Km' plasmid in donor of GEM and N1 strains wah similar a.;about 10 ' when co~ljugation was conducted wit11 M'I'I strain is recipient at pH 7. butthat of 1)KCOOI was lowered to 1.2X 10 '. And when the lab. stlain was uhccl as recipient.the transfer tendency of the plasmid was about same in both (;EM and NI strains usedas donor. All thc tionor 5trains. except for I)KC601. showecl the Ilighcs~ frequency of about10 ' at pH 7 and the frequcncics were lowered at both pH 5 and 9. Hut the mocliliedKm' plasmid in the cloned strain of DKC601 was transferred hy very low frequency of10 "at pH 5 ant1 7 comparing to other GEM strains. especiall! any co~~.jugantws ere notobtained at pH 4 and 9 even after conjugation for 6 hours. Rearrangement of the plasmidstranskrred into the lab. strain was not found in the conjugants. I\ulcornerut a lot of rearrangclncntwas ohservecl nlhen they were transferred into the NI strain. Such a rearrangement wasmore severe when donor was GEM strain rather then NI strain Hut such ;r phenomenonwas less affected by p!-l values.r phenomenon was less affected by p!-l values.

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Comparative Genetic Characterization of Plasmids of Agrobacterium Species Isolated in Korea (한국산 Agrobacterium plasmid의 유전학적 성상에 관한 연구)

  • Kim, Jung-Hye;Koo, Yong-Bum;Lee, Ki-Yung;Chung, Jae-Kyu
    • Journal of Yeungnam Medical Science
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    • v.1 no.1
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    • pp.41-48
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    • 1984
  • The soil bacterium Agrobacterium tumefaciens is a plant pathogen that cause3 crown gall tumors by infecting the wounded dicotyledonous plants and subsequent integration of bacterial DNA into plant nuclear DNA. Virulent A. tumefaciens strains harbor a large Ti (tumor-inducing) plasmid that carries genes essential for tumorigenesis. In the present study, 13 strains (Malus pumila Mill; $A_{1-3}$, Populus monilifera; $W_{1-6}$, Populus tomentiglandlosa; $P_{1-3}$ and Rosa species; $R_1$) of Agrobacterium isolated in korean crown gall tumors and plasmids were observed in 6 strains ($W_2$, $W_3$, $W_6$, $P_1$, $P_3$ and $A_2$). The test for crown gall tumor formation was resulted only in ATCC15955 and $KW_2$ strains inoculated into the stem of sun flower and the development was observed for 4 and 6 weeks after inoculation. Above two Ti plasmids (pTi) were purified by cesium chloride-ethidium bromide density gradient centrifugation and digested with restriction enzyme and fragments of pTiATCC 15955 and $pTiKW_2$ observed by EcoR I ; 25&27, Hind III; 23&21, BamH I ; each 20 and Hpa I ; 12&27, and sizes of pTiATCC15955 and and $pTiKW_2$ calculated as 200 and 87 kbases. Octopine was isolated from tumor tissue ($W_{1-6}$ and $P_{1-3}$) and these strains confirmed as octopine type.

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Drug Resistance and R Plasmids of Escherichia coli in Patients and Healthy Individuals in Korea (한국(韓國)의 환자(患者) 및 건강인(健康人)에서 분리(分離)한 E.coli의 약제내성(藥劑耐性) 및 R Plasmids)

  • Seol, Sung-Yong
    • The Journal of the Korean Society for Microbiology
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    • v.12 no.1
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    • pp.11-18
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    • 1977
  • A total of 665 strains of Escherichia coli isolated in Korea from stools of patients who were treated with antimicrobial drugs, doctors, and students were tested for the drug resistance and distribution of R plasmids. Approximately 25 to 41% of isolates were resistant to chloramphenicol, tetracycline, streptomycin, sulfisemidine and ampicillin(AP), and 9.5% were resistant to kanamycin. Nalidixic acid-resistant strains were only rarely encountered. The prevalence of resistant strains was significantly higher among patients than doctors and students. Strains multiply resistant to four or more drugs were significantly more prevalent among patient isolates than those from doctors and students, while no difference on the incidence of strains resistant to three or less drugs was noted among isolates from the three groups. The persons carrying strains resistant to four or more drugs were more frequently found among patients than doctors and students. Quite large proportions of drug-resistant strains transferred their resistance to drug-sensitive E. coli, with frequent transfer of whole resistance and AP resistance. Strains having higher multiplicity of resistance showed a tendency toward higher incidence of resistance transfer, irrespective of the origins of strains.

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Role of CopA to Regulate repABC Gene Expression on the Transcriptional Level (전사 수준에서 repABC 유전자 발현을 조절하는 CopA 단백질의 역할)

  • Sam Woong Kim;Sang Wan Gal;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;In Gyu Baek;Kyu Ho Bang
    • Journal of Life Science
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    • v.34 no.2
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    • pp.86-93
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    • 2024
  • Since replication of plasmids must be strictly controlled, plasmids that generally perform rolling circle replication generally maintain a constant copy number by strictly controlling the replication initiator Rep at the transcriptional and translational levels. Plasmid pJB01 contains three orfs (copA, repB, repC or repABC) consisting of a single operon. From analysis of amino acid sequence, pJB01 CopA was homologous to the Cops, as a copy number control protein, of other plasmids. When compared with a CopG of pMV158, CopA seems to form the RHH (ribbon-helix-helix) known as a motif of generalized repressor of plasmids. The result of gel mobility shift assay (EMSA) revealed that the purified fusion CopA protein binds to the operator region of the repABC operon. To examine the functional role of CopA on transcriptional level, 3 point mutants were constructed in coding frame of copA such as CopA R16M, K26R and E50V. The repABC mRNA levels of CopA R16M, K26R and E50V mutants increased 1.84, 1.78 and 2.86 folds more than that of CopA wt, respectively. Furthermore, copy numbers owing to mutations in three copA genes also increased 1.86, 1.68 and 2.89 folds more than that of copA wt, respectively. These results suggest that CopA is the transcriptional repressor, and lowers the copy number of pJB01 by reducing repABC mRNA and then RepB, as a replication initiator.