• 제목/요약/키워드: Restriction Enzyme Mapping

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황색포도상구균에서 테트라사이클린 내성을 나타내는 플라스미드의 동정 (Characterization of Tetracycline Resistant Plasmid in Staphylococcus aureus by Restriction Enzyme Mapping)

  • 김기현;김종명;문경호
    • 약학회지
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    • 제36권3호
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    • pp.255-258
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    • 1992
  • The clinical isolate Staphylococcus aureus SA8 was resistant to tetracycline(Tc) and harboured a plasmid pKH1(24.82 kb). pKH1 was shown by curing and by transformation to specify resistance to Tc. The cleavage map of a pKH1 was determined by restricction enzyme mapping techniques. Cleavage map is given for BglII, EcoRI, HpaII, PvuII and SalI. Restriction endonuclease BamHI, BglI, BstEII, HpaI, PstI, and XhoI have no sites on this plasmid. HaeIII, XbaI, and HindIII have 5, 6, 14 sites, respectively.

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Pseudomonas syringe pv. phaseolicola로 부터 제한효소의 분리정제 및 특성 (Purification and Characterzation of a Restriction Endonuclease from Pseudomonas syringae pv.phaselicola)

  • 배무;이은영
    • 한국미생물·생명공학회지
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    • 제22권5호
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    • pp.485-490
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    • 1994
  • A restriction endonuclease, PsyI, has been isolated from Pseudomonas syringae pv. pha- seolicola, and its catalytic properties have been studied. This enzyme was purified through strepto- mycin sulfate and ammonium sulfate fractionation, phosphocellulose Pll, DEAE-cellulose, hydroxy- apatite and Sephadex G-100 column chromatography. It's molecular weight was about 50,000 dalton as determined by 7.5% polyacrylamide gel electrophoresis containing 0.1% SDS. In catalytic proper- ties, PsyI shows stable at wide ranges of pH between 7.0 and 10.0, of temperature between 30$\circ$C and 37$\circ$C, and its thermal stability is between 25$\circ$C, and 45$\circ$C, at the presence Of 10 mM MgCl$_{2}$-PsyI essentially require Na salt for enzyme reaction, is rather inhibited in the high Na salt concent- ration. The presence of 2-mercaptoethanol is absolutely required for the enzyme activity. This endonuclease, PsyI was determined to be an isoschizomer of SalI from the results of the restriction mapping and DNA sequencing.

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Subcloning of Nodulin 26 Wild Type(S262) and Phosphorylation Site Mutant(S262D) into the Yeast Expression Vector pYES2

  • Cha, Youn-Soo
    • Preventive Nutrition and Food Science
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    • 제2권1호
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    • pp.61-65
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    • 1997
  • Wild type nodulin 26(nod 26) cDNA(S262) and phodphorylation aite mutant(S262D) were constructed by a yeast expression system using pYES2 plasmids(pTES2-D262 and pTES2-S262D) were sc-reened by restriction mapping with BamHI of KpnI. S262 nod 26 contained a sreine residue at position 262 and S262D nod 26 contained the substitution mutation of serine to aspartic acid residue at position 262 were verified by automated floursent DNA sequencing.

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생명공학 단원의 제한 효소 지도 작성 탐구실험 수업이 고등학생의 과학긍정경험에 미치는 영향 (The Effects of Positive Experience about Science of High School Students in an Inquiry Experiment Class on Restriction Enzyme Mapping in Biotechnology Chapter)

  • 정수연;장정호
    • 과학교육연구지
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    • 제46권3호
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    • pp.293-311
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    • 2022
  • 본 연구는 분자생물학 기초 소양 함양을 위한 제한 효소 지도 작성의 탐구실험을 구성하였고, 일반계 고등학교 2학년을 대상으로 학생 중심의 제한 효소 지도 작성 탐구실험 수업을 통해 탐구실험 능력과 과학긍정경험에 미치는 영향을 분석하였다. 우선, 탐구실험 수업이 일반계 고등학생의 과학긍정경험에 있어 '그렇다' 이상으로 응답한 비율이 사전에 비해 사후에서 높은 것으로 보아 유의미하게 효과가 있는 것으로 나타났다. 제한 효소 지도 작성을 위한 연속의 5단계로 구성하여 적용한 결과 학생들의 과학 분야 학업에 대한 흥미 유발뿐만 아니라 학생들의 수업 참여도가 높아졌고, 구체적인 과학 학습 동기, 과학 진로 포부 및 체험 자료로도 효과적인 것으로 나타났다. 탐구실험 수업의 과학 환경이 학생들의 학습 태도와 과학긍정경험의 향상으로 이어져 수업의 집중과 수업의 질적 향상, 학생간의 적극적인 의사 소통과 상호 협력의 중요성에 긍정적인 영향을 끼쳤다. 또 탐구실험 수업이 진로 체험의 기회를 제공함으로써 분자생물학의 기초 소양 함양과 이공계 진학의 토대가 될 것이다.

다제내성 황색포도상구균이 가지고 있는 테트라사이클린 내성 플라스미드의 동정 (Characterization of Tetracycline Resistance Plesmid of Multidrug-resistant Staphylococcus aureus)

  • 이대운;문경호
    • 약학회지
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    • 제39권1호
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    • pp.6-9
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    • 1995
  • The clinical isolate Staphylococcus aureus SA2 had four kinds of plasmids and was resistant to ampicillin, chloramphenicol, clindamycin, erythromycin, gentamicin, kanamycin, methicillin, streptomycin, tetracycline and tobramycin. Transformation experiment demonstrated that 4.44 kb plasmid(pKH6) encoded resistance to tetracycline. The cleavage map of pKH6 was determined by restriction enzyme mapping techniques. The cleavage map is given for EcoRV, HindIII, HpaI, HpaII, KpnI and Xbal. Restriction endonucleases BamHl, BglI, BGIII, BstEII, EcoRI, HaellI, PstI, PvuII, SalI, Smal, and Xhol have no site on this plasmid. The restriction map revealed extensive structural homology between pKH6 and pT181.

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다제내성 황색포도상구균이 가지고 있는 클로람페니콜 내성 플라스미드의 동정 (Characterization of Chloramphenicol Resistant Plasmid of Multidrug-resistant Staphylococcus aureus)

  • 이대운;문경호
    • 약학회지
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    • 제37권6호
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    • pp.621-624
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    • 1993
  • The clirical isolate Staphylococcus aureus SA2 had four kinds of plasmids and was resistant to ampicillin, chloroamphenicol, clindamycin. erythromycin, gentamicin, kanamycin, methicillin, streptomycin, tetracycline and tobramycin. Transformation experiment demonstrated that 4.14kb plasmid(pKH7) encoded resistance to chloramphenicol. The cleavage map of pKH7 was determined by restriction enzyme mapping techniques. The cleavage map is given for BstEll, Hindlll, Hpall, and Xbal. The above restriction endonucleases have a single site, but nucleases BamHl, Bgll, BglII, EcoRl, EcoRV, HaeIII, Hpal, Kpnl, Pstl, PvnII, Sall, Smal, and XhoI have no site on this plasmid.

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Klebsiella pneumoniae NFB-320의 Pullulanase 유전자의 제한효소 분석과 효소학적 특성 (Restriction Mapping of Cloned Pullulanase Gene and Property of Pullulanase Produced in Escherichia coli (pYKL451) and Klebsiella pneumoniae NFB-320)

  • Yu, Ju-Hyun;Chung, Kun-Sub;Kong, In-Su;Lee, Jung-Kee
    • 한국미생물·생명공학회지
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    • 제15권6호
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    • pp.436-440
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    • 1987
  • 앞서 보고한 바와 같이 토양으로부터 분리한 K. pneumoniae NFB-320의 pullulanase 유전자를 pBR 322을 이용하여 E. coli에 cloning한 결과 약 14.4kb 의 재조합 plasmid DNA pYKL451을 얻었다. 이러한 pullulanase 유전자에 대한 유전적 정보를 얻기 위해 여러 가지 제한효소로 단일 혹은 이중 절단을 행하여 삽입된 pullulanase 유전자의 제한효소 절단지 도를 작성하였으며, E. coli(pYKL451)과 K. pneumoniae NFB-320이 생산하는 pullulanase의 효소적 특성을 조사하였다. 생산되는 두 균주의 효소는 50-55$^{\circ}C$ 부근에서 최적온도를 나타냈으며 최적pH는 모두 6.0이었다. 효소 안정성에 미치는 pH의 영향은 4$0^{\circ}C$에서 90min 간 방치했을 때 pH 5.0-10.0에서 안정하였으며 열안정성은 (pH6.0) 각 온도에서 한시간 처리하였을 때 4$0^{\circ}C$까지는 안정하였으나 5$0^{\circ}C$ 이상에서는 효소의 활성이 급격히 감소하였다.

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A Molecular Marker Specific to Metarhizium anisopliae var. majus

  • YOON, CHEOL-SIK;GI HO SUNG;JAE MO SUNG;JAEANG OON LEE
    • Journal of Microbiology and Biotechnology
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    • 제9권3호
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    • pp.334-339
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    • 1999
  • More innovative molecular markers were investigated for rapid and consistent differentiation of Metarhizium anisopliae var. majus from M. anisopliae var. anisopliae. A total of 28 isolates were obtained from various countries and hosts: 13 isolates of M. anisopliae var. anisopliae, 12 isolates of M. anisopliae var. majus, and 3 isolates of M. anisopliae collected in Korea. This study involved restriction enzyme digestions of a PCR product amplified from nuclear internally transcribed spacer (ITS) and a portion of the 28S rDNA regions. Among 11 different restriction enzymes used in this study, MboⅠ digestion particularly produced a restriction pattern that had characteristics of M. anisopliae var. majus. This restriction pattern was consistent in all isolates of M. anisopliae var. majus regardless of their geographic origins and insect hosts. Mapping experiments revealed that MboⅠ sites of M. anisopliae var. majus are identical to those of M. anisopliae var. anisopliae with an exception for the presence of an additional site in the PCR product. Results from this study provide an additional method for identification and differentiation of isolates of these two varieties of M. anisopliae for use in the field and laboratory experiments.

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SEQUENCE ANALYSIS AND COMPARISON OF BOVINE αS1-CASEIN GENOMIC DNA

  • Lin, C.S.;Huang, M.C.;Choo, K.B.;Tseng, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제6권4호
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    • pp.541-547
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    • 1993
  • A phage clone containing the partial ${\alpha}_{S1}$-casein gene was isolated from a bovine genomic library by using mixed probes of ovine ${\alpha}_{S1}$-, ${\beta}$- and ${\kappa}$-casein cDNAs. Restriction enzyme mapping analysis for 14.6 kb revealed that the map was in conflict with the report of Meade et al. (1990), especially in the 3'-end fragment. Sequence analysis of 12.6 kb revealed a high AT/GC ratio (1.64); we have identified eight exon sequences according to the bovine ${\alpha}_{S1}$-casein cDNA sequence. The same exon/intron splice junction sequence was observed between these exons. We suggest that the bovine ${\alpha}_{S1}$-casein gene night contain a minimum of 18 exons and the full length is approximately 18-19 kb.

황색포도상구균의 항생제 다제내성을 갖는 플라스미드의 동정 (Characterization of Multidrug Resistant Plasmid of Staphylococcus aureus)

  • 김기현;이대운;김종명;문경호
    • 약학회지
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    • 제36권5호
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    • pp.486-490
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    • 1992
  • The clinical isolate Staphylococcus aureus SA2 was resistant to ampicillin, Chloramphenicol, clindamycin, erythromycin, gentamicin, kanamycin, methicillin, streptomycin, and tobramycin and harboured more than two kinds of plasmids. Transformation experiment demonstrated that 40.98-kb plasmid(pKH2) encoded resistance to ampicillin, clindamycin, erythromycin, kanamycin, and streptomycin. The cleavage map of a pKH2 was determined by restriction enzyme mapping techniques. Cleavage map is given for BamHI, BglI, BstEII, SalI and XhoI.

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