• Title/Summary/Keyword: kanamycin-resistant

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Expression of Pea Superoxide Dismutase Gene in Transgenic Cucumber (Cucumis sativus L.) Plants (형질전환 오이(Cucumis sativus L.) 식물체에서 완두 Superoxide Dismutase 유전자의 발현)

  • 김재훈;오승용;이행순;조만현;이은모;우인식;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.201-206
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    • 1998
  • To develop the fruits of cucumber (Cucumis sativus L.) producing high yields of superoxide dismutase (SOD), the MnSOD cDNA from pea (Pisum sativum) under the control of the cauliflower mosaic virus 35S promoter was introduced into cucumber using Agrobacterium tumefaciens (strain LBA 4404)-mediated transformation. The kanamycin-resistant shoots were selected on the selection medium containing MS basal salt, 1.0 mg/L zeatin, 0.1 mg/L IAA, 300 mg/L claforan, and 100 mg/L kanamycin. After 6 weeks of culture on the selection medium, the shoots were transferred to MS medium containing 0.2 mg/L NAA to induce roots. PCR analysis using the primers for neomycin phosphotransferase (NPTII) gene revealed that three plantlets were transformed. The fruits of one transgenic plant had approximately 3.2-fold higher SOD activity than those of non-transgenic plants. MnSOD isoenzyme band was strongly detected on native gel in fruits of transgenic plants.

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Transformation of Populus alba $\times$Populus glandulosa Using Phosphinothricin Acetyltransferase Gene (Phosphinothricin acetyltransferase 유전자를 이용한 현사시의 형질전환)

  • 오경은;양덕춘;문흥규;박재인
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.163-169
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    • 1999
  • This study was conducted to produce herbicide resistant plants by transferring phosphinothricin acetyltransferase (PAT) gene into Populus alba $\times$ Populus glandulosa No .3 using Agrobacterium tumefaciens MP 90/PAT. Leaf segments from in vitro grown shoots of hybrid poplar No. 3 were soaked in a AB medium containing Agrobacterium tumefaciens MP 90/PAT for 10 min and cocultivated for 2 days on MS medium containing 1.0 mg/L 2,4-D and 0.2mg/L kinetin (CIM). Putative transformed calli could be selected after cocultivation of leaf segments on CIM supplemented with 50mg/L kanamycin and 500mg/L cefotaxime for 3 weeks. The selected calli were cultured on CIM supplemented with 50 mg/L kanamycin and 500 mg/L cefotaxime for 5~8 weeks before transfer to WPM containing 1.0mg/L zeatin, 0.1mg/L BAP, 50 mg/L kanamycin and 500mg/L cefotaxime for shoot regeneration. Shoots were regenerated from the callus after 4 week cultivation, and the regenerants were grown on the same medium for 7~l0 weeks. The plants rooted on 1/2 WPM containing 0.2 mg/L IBA and 50 mg/L kanamycin. To confirm the gene insertion into plants, GUS activity was detected by histochemical assay in the transformed plants. Finally, the presence of both NPT II and PAT genes from the transgenic plants were confirmed by PCR amplification with the gene specific primers and subsequent PCR-Southern blot with DIG-labeled PAT gene probe. After acclimatization in pots for 4 weeks, the plants were sprayed by 3 mL/L of Basta to test resistance to the herbicide. The transgenic plants remained green, whereas all the control plants died after one week.

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Antibiotic Susceptibility of Salmonella spp. Isolated From Diarrhoea Patients in Seoul From 1996 to 2001 (서울 시내 설사환자에서 분리한 살모넬라의 항생제 감수성의 년도별 변화 추이)

  • 박석기;박성규;정지헌;진영희
    • Journal of Food Hygiene and Safety
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    • v.17 no.2
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    • pp.61-70
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    • 2002
  • In order to investigate the classification and antibiotic resistance of Salmonella species,718 isolates were isolated from patient in Seoul from 1996 to 2001. The two hundred and ninety eight isolates (41.5%) were identified as Sal. Enteritidis, followed by Sal. Typhi 218 isolates (30.4%), and Sal. Typhimurium 87 isolates (12.1%). The identified Salmonella species were most resistant to tetracycline (32.7%), followed by streptomycin (28.0%), ticarcillin (18.1%) and ampicillin (12.4%). Among isolates,34.7% of Sal. Enteritidis were resistant to tetracycline, 32.3% to streptomycin,23.2% to ticarcillin,13.5% to ampicillin, respectively. 13.8% of Sal. Typhi were resistant to streptomycin,10.6% to tetracycline, respectively.66.7% of Sal. Typhimurium were resistant to tetracycline, 42.5% to streptomycin, 28.7% to ticarcillin, 26.4% to ampicillin and 17.2% to chloramphenicol, respectively. Of 718 isolates, 324 isolates (45.1%) were resistant to 1 or more drugs and 64 isolates (19.8%) were resistant to 1 drug, 132 isolates (40.7%) were resistant to 2 drugs,50 isolates (15.4%) were resistant to 3 drugs, 27 isolates (8.3%) to 4 drugs,27 isolates (8.3%) to 5 drugs,22 Isolates (6.8%) to 6 drugs. The most prevalent multiple resistant pattern was tetracycline-kanamycin (35.5%), followed by tetracycline-kanamycin-ticarcillin (8.3%), and tetracycline-kanamycin-ticarcillin-ampicillin (7.4%) . Antibiotic resistant rate of Sal. Typhimurium was 73.6%,1311owe4 by Sal. Enteritidis 53.7% and Sal. Typhi 19.3%. Most Sal. Enteritidis was resistant to 1 drug o.2 drugs, whereas Sal. Typhi. and Sal.. Typhunurium were more .resistant to 5 (16.7%) or 6 drugs (26.6%). The old generation antibiotics such as ampicillin, tetracycline, and streptomycin were annually more resistant than the new generation antibiotics such as ceftriaxone, ciprofloxacin or cefoxitin.

Antibiotic Resistance of Escherichia coli from Pigs Fed with Antibiotics-containing Feedstuffs (돈유래(豚由來) Escherichia coli의 항생물질내성(抗生物質耐性) 및 전달성내성인자(傳達性耐性因子)에 관(關)하여)

  • Tak, Ryun Bin;Chung, Gill Taik
    • Korean Journal of Veterinary Research
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    • v.16 no.2
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    • pp.159-163
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    • 1976
  • One hundred and sixty one Escherichia coli strains isolated from 24 swine (11 swine fed with feedstuffs containing 7.5mg/kg of tetracycline and 13 swine not received antibiotic) were studied for the drug resistance and distribution of R factors. About 42 per cent of E. coli strains isolated from pigs of a herd fed with tetracycline (TC)-containing feeds were resistant to TC, streptomycin(SM), sulfisomidine(SA), ampicillin (AP) and kanamycin (KM), alone or in combination thereof, but none of the swine not receiving antibiotic containing feedstuffs excreted E. coli resistant to these drugs, Among resistant strains, 18.2% were found to be singly resistant to TC, whereas 81.8% were resistant to two or more antibiotics. The most common pattern was the triple resistant to TC, SM and SA(30.3%), and follolwed by double ones to TC and SM(24.2%). About one half of resistant strains carried R factors which were tranferable to the recipients by conjugation. In spite of feeding with feedstuffs containing only TC, high incidences of multiple resistance and R factors were observed in the E. coli isolated from these swine.

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Annual Population Variation and Identification of Antibiotic-Resistant Bacteria in the Lower Lake Geumgang (금강호의 항생제 내성세균의 분포 및 동정)

  • Bae, Myoung-Sook;Choi, Gang-Guk;Park, Suhk-Hwan;Choi, Moon-Sul;Lee, Geon-Hyoung
    • The Korean Journal of Ecology
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    • v.27 no.5
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    • pp.283-289
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    • 2004
  • This study was conducted to evaluate the annual population variation and identification of antibiotic-resistant bacteria in the lower artificial Lake Geumgang from January to December, 2002. Samples were taken from the surface waters at 3 stations near the estuarine barrage. The results were as follows; the population densities of heterotrophic bacteria varied from 4.1±1.0×10² to 6.7±1.1×10³ cfu ml/sup -1/ during the investigation periods. The population densities of antibiotic-resistant bacteria ranged from 1.5±0.7×10 to 4.3±0.3×10³ cfu ml/sup -1/ for ampicillin; from 0 to 6.4±0.4× 10² cfu ml/sup -1/ for chloramphenicol; from 0 to 2.8±0.3×10³ cfu ml/sup -1/ for gentamicin; from 0 to 4.5±1.0×10³ cfu ml/sup -1/ for kanamycin; and from 1.0±0.4 × 10 to 2.3±0.5×10³ cfu ml/sup -1/ for streptomycin, respectively. Of the sixty isolates, 90% were Gram negative. Dominant genera by 16S rDNA analysis were identified Aeromonas spp. (14 strains), Bacillus spp. (6 strains), Enterobacter spp. (4 strains), and Stenotrophomonas spp. (6 strains). These strains were clustered into 12 groups based on relatedness by average linkage method. Of the 60 isolates, 85% had the resistance to ampicilin and 32% were shown resistance to more than 2 kinds of antibiotics.

Introduction of Bean Chitinase Gene into Korean Ginseng by Agrobaterium tumefaciens (Agrobacterium tumefaciens에 의한 강낭콩 키틴가수분해효소 유전자의 고려인삼으로의 도입)

  • 이행순;권석윤;백경희;김석원;이광웅;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.95-99
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    • 1995
  • We have previously established a system for plant regeneration through somatic embryogenesis and Agrobacterium-mediated transformation of Korean ginseng. In this study to produce a fungus-resistant plant, we introduced a bean chitinase gene into ginseng using the transformation system. A binary vector pChi/748 was constructed by introducing the bean basic chitinase gene into EcoRI site of pGA748 which carries the CaMV 35S promoter governing the introduced gene and neomycin phosphotransferase II(NPT-II)gene as a positive selection marker. Cotyledonary explants were cocultured with A. tumefaciens strain LBA4404 harboring the binary vertor pChi/748 for 48 h, and transferred to MS medium supplemented with l mg/L2,4-D,0.1mg/L kinetin, 100 mg/L kanamycin, and 500mg/L carbenicillin. Kanamycin-resistant calli were formed on the cut surface of cotyledonary explants after one month of culture, and subsequently they gave rise to somatic embryos. Upon transfer onto medium containing 1 mg/L each of BA and GA$_3$, most of them converted to plantlets after 5 weeks of culture. The genomic DNA of eight kanamycin-resistant regenerants was subjected to polymerase chain reaction (PCR) using two specific 21-mer oligonucleotides derived from the chitinase gene. PCR-Southern blot analysis confirmed that the chitinase gene was incorporated into six out of the eight regenerants..

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Expression of in vitro-tested ribozyme against cucumber mosaic virus RNA in tobacco plant (시험관내에서 합성한 오이모자이크 바이러스 RNA단편을 성공적으로 절단한 ribozyme의 식물체내의 발현)

  • Park, Sang-Gyu
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.355-360
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    • 1996
  • In vitro-tested ribozyme against synthesized cucumber mosaic virus (CMV) RNA (Agric. Chem. & Biotech. 37:56-63(1994)) was expressed in tobacco plant to develop virus resistant plants. The ribozyme sequence was linked to cauliflower mosaic virus 35S promoter and nopaline synthase(nos) terminator and this chimeric 35S-ribozyme-nos gene was sequenced. The sequenced chimeric gene was transferred to Agrobacterium tumefaciens LBA4404 using tri-parental mating system. The E. coli HB101 containing chimeric gene was incubated with E. coli HB101(pRK2073) as a helper and Agrobacterium tumefaciens LBA4404. Then Agrobacterium cells containing the ribozyme construct was cocultivated with tobacco leaf pieces. Ten different plants were regenerated from kanamycin containing MS medium. The presence of the ribozyme construct in the transgenic tobacco plants was confirmed by polymerase chain reaction (PCR). Seven different transgenic plants in ten different kanamycin resistant plants showed the expected size (570 base pairs) of 35S-ribozyme-nos gene fragment. Total RNAs were isolated from four different transgenic plants and separated on a 1% agarose gel containing formamide. Northern hybridization with 35S-ribozyme-nos gene fragment as a probe indicated that ribozyme transcripts may be degraded tv nuclease. Therefore, nuclease-resistant ribozymes are needed for the development of virus-resistant transgenic plants using ribozymes.

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Exchange method of the nasal cavity spraying amikacin instead of kanamycin and gentamicin (가나마이신 및 겐타마이신 비강 분무액을 아미카신 비강 분무액으로 바꾸는 방법)

  • Kim, Hong-Jib
    • Korean Journal of Veterinary Research
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    • v.49 no.2
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    • pp.113-119
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    • 2009
  • Atrophic rhinitis (AR) is the one of important respiratory diseases and causes severe economic losses in pig industry. Severe attempts have been made to reduce the economic losses by preventing the disease. One of the methods is the spraying of antibiotics into nasal cavity of piglets. Recently, the efficacy of the spraying with kanamycin and gentamicin was reduced in the Korean swine industry. Therefore, the preventive methods have been required to be changed based on the antimicrobial susceptibility profiles of causative agents of swine AR. Based on the current situations of this disease, Bordetella (B.) bronchiseptica and Pasteurella (P.) multocida 4D were isolated from pigs with clinical signs of AR. The isolation rates of B. bronchiseptica and P. multocida 4D were 58.5% and 32.9%, respectively. In the antimicrobial susceptibility test, the bacteria were resistant to kanamycin and gentamicin which have been used as the spraying agents, but they were susceptible to amikacin. A new spraying agent was developed using amikacin using ${\beta}$-glucan and yakbaltag as supplementary agents. Field efficacy of the agent was carried out with different schedule. The results from this study suggested that the newly developed spraying agents might be helpful to prevent AR in swine.

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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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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