• Title/Summary/Keyword: Restriction Enzyme Mapping

Search Result 29, Processing Time 0.021 seconds

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

  • Kim, Ki-Hyun;Kim, Jong-Myung;Moon, Kyung-Ho
    • YAKHAK HOEJI
    • /
    • v.36 no.3
    • /
    • pp.255-258
    • /
    • 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.

  • PDF

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

  • Bae, Moo;Lee, Eun-Young
    • Microbiology and Biotechnology Letters
    • /
    • v.22 no.5
    • /
    • pp.485-490
    • /
    • 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.

  • PDF

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
    • /
    • v.2 no.1
    • /
    • pp.61-65
    • /
    • 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.

  • PDF

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

  • Soo Yeon Jeong;Jeong Ho Chang
    • Journal of Science Education
    • /
    • v.46 no.3
    • /
    • pp.293-311
    • /
    • 2022
  • In this study, a restriction enzyme mapping inquiry experiment was developed for cultivating basic knowledge on molecular biology and the effects on inquiry experiment ability and positive experience on science through student-centered molecular biology inquiry experiment class for second graders of a general high school was analyzed. First of all, it was found that the experimental class through the inquiry experiment was significantly effective as the percentage of high school students who answered 'yes' or higher in the positive science experience of general high school students was higher after than before the test. As a result of developing and applying a series of five classes for the creation of restriction enzyme maps, not only did the students' interest in science studies, but also their class participation increased. They were also used as effective specific science learning motives, science career aspirations and experience data. The science environment of the inquiry experiment class led to the improvement of students' learning attitudes and positive science experience, which had a positive effect on the importance of class concentration and class quality, active communication and mutual cooperation among students. In addition, inquiry and experiment classes will provide opportunities for career experience, which will become the foundation for cultivating basic knowledge on molecular biology and advancing to science and engineering.

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

  • 이대운;문경호
    • YAKHAK HOEJI
    • /
    • v.39 no.1
    • /
    • pp.6-9
    • /
    • 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.

  • PDF

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

  • 이대운;문경호
    • YAKHAK HOEJI
    • /
    • v.37 no.6
    • /
    • pp.621-624
    • /
    • 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.

  • PDF

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

  • Yu, Ju-Hyun;Chung, Kun-Sub;Kong, In-Su;Lee, Jung-Kee
    • Microbiology and Biotechnology Letters
    • /
    • v.15 no.6
    • /
    • pp.436-440
    • /
    • 1987
  • Pullulanase gene (pul) of Klebsiella pneumoniae NFB-320 which was cloned previously in Escherichia coli with plasmid pBR322. The gene was analyzed with various restriction enzymes. The cloned gene was contained within n 10 kb BamHI DNA fragment. We constructed the restriction map of the hybrid plasmid pYKL451. The optimum temperatures for pullulanases produced in E. coli (pYKL451) and K. pneumoniae NFB-320 were almost the same, 50-55 $^{\circ}C$. The optimum pHs for the reaction of the enzymes produced by E. coli (pYKL451) and K. pneumoniae NFB-320 was 6.0. Both enzyme preparations were stable under the range of pH 5.0 to 10.0 when those were kept at 40 $^{\circ}C$ for 90 min and were stable until 40 $^{\circ}C$ when allowed to stand for 1hr at various temperatures.

  • PDF

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
    • /
    • v.9 no.3
    • /
    • pp.334-339
    • /
    • 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.

  • PDF

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
    • /
    • v.6 no.4
    • /
    • pp.541-547
    • /
    • 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.

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

  • Kim, Ki-Hyun;Lee, Dae-Woon;Kim, Jong-Myung;Moon, Kyung-Ho
    • YAKHAK HOEJI
    • /
    • v.36 no.5
    • /
    • pp.486-490
    • /
    • 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.

  • PDF